Promoters for driving gene expression in rod photoreceptors and uses thereof

Abstract

This disclosure provides promoters and expression construct and vectors comprising the promoters for expression of operatively linked transgene in rod photoreceptor calls. Also provided are methods of using the rod-specific promoters to treat rod disorders.

Description

PROMOTERS FOR DRIVING GENE EXPRESSION IN ROD PHOTORECEPTORS AND USES THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 U.S.C. §119(e) of the earlier filing date of U.S. Provisional Patent No. 63 / 709,883, filed on October 21, 2024, which is hereby incorporated by reference in its entirety.REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0002] The contents of the electronic sequence listing (162027.53876SeqList.xml; Size: 149,115 bytes; and Date of Creation: September 26, 2025) is herein incorporated by reference in its entirety.FIELD

[0003] The present disclosure relates generally to the field of molecular biology and medicine. More particularly, the methods and compositions herein are useful for applications involving gene therapy.BACKGROUND

[0004] The retina is a layer of photoreceptors and glial cells within the eye. This layer is responsible for capturing incoming photons and transmitting them to the brain to perceive a visual image. Photoreceptor cells comprise rod photoreceptors and cone photoreceptors. Rod cells contain rhodopsin (made of retinal, which is a vitamin A aldehyde), which is a light sensitive pigment that enables the absorption of photons. As a consequence, rods are highly sensitive to light, which enables them to perceive a small amount of light in dim conditions (z.e., rods enable scotopic vision). However, rods are saturated by light intensities that are too high. In contrast, cones, which are responsible for color vision, are significantly less lightsensitive. As such, cones can provide for vision even in bright light (z.e., cones enable photopic vision). Further, cones provide a higher visual acuity and temporal resolution than rods. Humans have three types of cones: blue-, green- and red-sensitive cones.

[0005] Cones are generally located in the center of the retina at the fovea (the central portion of the macula that is responsible for central vision). In contrast, rods are present at high densitythroughout most of the retina, with a sharp decline in the fovea. Rod photoreceptors outnumber cone photoreceptors by a ratio of 20: 1 or greater in most mammalian retinas.

[0006] Mutations that effect rod function can lead to various ocular diseases and are associated with various rod disorders including inherited retinal diseases (IRDs). For example, one common IRD is caused by mutations in the rhodopsin gene, which are associated with autosomal dominant retinitis pigmentosa (adRP).

[0007] Unfortunately, to date, there has been little progress in designing strong, rodspecific promoters particularly for expressing therapeutic transgenes in rod photoreceptor cells. As such, the development of gene therapy approaches specifically targeting rod photoreceptors has been hampered. Accordingly, new rod-specific promoters for use in ocular gene therapy are urgently needed.SUMMARY

[0008] Provided herein are promoters, and expression constructs comprising these promoters, for expression of transgenes in rod photoreceptors. Also provided are vectors comprising the promoters and expression constructs described herein, and pharmaceutical compositions comprising such constructs or vectors. Also provided are methods of using such constructs, vectors, and pharmaceutical compositions. Provided herein are promoters that are useful for driving the rod-specific expression of an operably linked transgene.

[0009] In one aspect, provided is a promoter, or an expression construct comprising the promoter, wherein the promoter comprises:(a) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 18 and (ii) a sequence that is at least 80% identical to SEQ ID NO: 17; or(b) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 19 and (ii) a sequence that is at least 80% identical to SEQ ID NO: 17.

[0010] In one aspect, provided is a promoter, or an expression construct comprising the promoter, wherein the promoter comprises:(a) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 18 and (ii) a sequence that is at least 80% identical to SEQ ID NO: 17; or(b) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 19 and (ii) a sequence that is at least 80% identical to SEQ ID NO: 17.

[0011] In some embodiments, the promoter comprises:(a) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 18 and (ii) a sequence that is at least 90% identical to SEQ ID NO: 17; or(b) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 19 and (ii) a sequence that is at least 90% identical to SEQ ID NO: 17.

[0012] In some embodiments, the promoter comprises:(a) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 18 and (ii) a sequence that is at least 90% identical to SEQ ID NO: 17; or(b) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 19 and (ii) a sequence that is at least 90% identical to SEQ ID NO: 17.

[0013] In some embodiments, the promoter comprises:(a) SEQ ID NO: 18 and SEQ ID NO: 17; or(b) SEQ ID NO: 19 and SEQ ID NO: 17.

[0014] In some embodiments, the promoter comprises:(a) SEQ ID NO: 18 and SEQ ID NO: 17; or(b) SEQ ID NO: 19 and SEQ ID NO: 17.

[0015] In some embodiments, the promoter comprises:(a) a nucleic acid sequence that is at least 80% identical to SEQ ID NO:8; or(b) a nucleic acid sequence that is at least 80% identical to SEQ ID NO:9.

[0016] In some embodiments, the promoter comprises:(a) a nucleic acid sequence that is at least 90% identical to SEQ ID NO:8; or(b) a nucleic acid sequence that is at least 90% identical to SEQ ID NO:9.

[0017] In some embodiments, the promoter comprises SEQ ID NO:8 or SEQ ID NO:9.

[0018] In some embodiments, the promoter consists of SEQ ID NO:8 or SEQ ID NO:9.

[0019] In one aspect, provided is a promoter, or an expression construct comprising a promoter, wherein the promoter comprises a nucleic acid sequence that is at least 80% identical to any one of SEQ ID NO: 10, SEQ ID NON, or SEQ ID NON. In some embodiments, the promoter comprises a nucleic acid sequence that is at least 90% identical to any one of SEQ ID NO: 10, SEQ ID NON, or SEQ ID NON. In some embodiments, the promoter comprises any one of SEQ ID NO: 10, SEQ ID NON, or SEQ ID NON. In some embodiments, the promoter consists of any one of SEQ ID NO: 10, SEQ ID NON, or SEQ ID NON.

[0020] In one aspect, provided is an expression construct comprising a promoter, wherein the promoter:(a) comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 1, wherein the promoter comprises no more than about 800 contiguous nucleotides from SEQ ID NO: 13;(b) comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO:5, wherein the promoter comprises no more than about 400 contiguous nucleotides from SEQ ID NO: 15;(c) comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO:6, wherein the promoter comprises no more than about 800 contiguous nucleotides from SEQ ID NO: 16; or(d) comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO:2, wherein the promoter comprises no more than about 400 contiguous nucleotides from SEQ ID NO: 14.

[0021] In some embodiments, the promoter:(a) comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 1 ; (b) comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO:5; (c) comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO:6; or(d) comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO:2.

[0022] In some embodiments, the promoter comprises any one of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:2.

[0023] In some embodiments, the promoter comprises or consists of a nucleic acid sequence that is at least 80% identical to any one of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:2. In some embodiments, the promoter comprises or consists of a nucleic acid sequence that is at least 90% identical to any one of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:2. In some embodiments, the promoter comprises or consists of any one of SEQ ID NO: 1, SEQ ID NO:5, SEQ ID NO:6, or SEQ IDNO:2.

[0024] In one aspect, provided is an expression construct comprising a promoter, wherein the promoter comprises:a) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 18, (ii) a sequence that is at least 80% identical to SEQ ID NO: 27, (iii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 28, and (iv) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 29;b) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 19, (ii) optionally SEQ ID NO: 27 (iii) a sequence that is at least 80% identical to SEQ ID NO: 28, and (iv) a sequence that is at least 80% identical to SEQ ID NO: 29; c) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 19, (ii) a sequence that is at least 80% identical to SEQ ID NO: 28, (iii) a sequence that is at least 80% identical to SEQ ID NO: 27, and (iv) a sequence that is at least 80% identical to SEQ ID NO: 30; ord) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 31, (ii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 28, and (iii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 32; or e) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 36, (ii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 21, and (iii) optionally the nucleic acid sequence CGCGGCCGC.

[0025] In some embodiments, the promoter comprises:a) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 18, (ii) a sequence that is at least 90% identical to SEQ ID NO: 27, (iii) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 28, and (iv) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 29;b) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 19, (ii) optionally, a sequence that is at least 90% identical to SEQ ID NO: 27, (iii) a sequence that is at least 90% identical to SEQ ID NO: 28, and (iv) a sequence that is at least 90% identical to SEQ ID NO: 29;c) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 19, (ii) a sequence that is at least 90% identical to SEQ ID NO: 28, (iii) a sequence that is at least 90% identical to SEQ ID NO: 27, and (iv) a sequence that is at least 90% identical to SEQ ID NO: 30; ord) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 31, (ii) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 28, and (iii) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 32; or e) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 36, (ii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 21, and (iii) optionally the nucleic acid sequence CGCGGCCGC.

[0026] In some embodiments, the promoter comprises:a) (i) SEQ ID NO: 18, (ii) SEQ ID NO: 27, (iii) SEQ ID NO: 28, and (iv) SEQ ID NO: 29;b) (i) SEQ ID NO: 19, (ii) SEQ ID NO: 27, (iii) SEQ ID NO: 28, and (iv) SEQ ID NO:29;c) (i) SEQ ID NO: 19, (ii) SEQ ID NO: 28, and (iii) SEQ ID NO: 29;d) (i) SEQ ID NO: 19, (ii) SEQ ID NO: 28, (iii) SEQ ID NO: 27, and (iv) SEQ ID NO:30; ore) (i) SEQ ID NO: 31, (ii) SEQ ID NO: 28, and (iii) SEQ ID NO: 32; orf) (i) SEQ ID NO: 36, (ii) SEQ ID NO: 21, and (iii) CGCGGCCGC.

[0027] In some embodiments, the promoter comprises:(a) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 22;(b) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 23;(c) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 24;(d) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 25; or (e) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 26.

[0028] In some embodiments, the promoter comprises:(a) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 22;(b) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 23;(c) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 24;(d) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 25; or (e) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 26.

[0029] In some embodiments, the promoter comprises any one of SEQ ID NOs: 3 and 22-26.

[0030] In one embodiment, the expression construct disclosed herein comprises a transgene operatively linked to a promoter described herein. In one embodiment, the transgene encodes a therapeutic protein. In some embodiments, the transgene encodes ArchT, R9AP (Rgs9-anchor protein or RGS9BP), Jaws (cruxhalorhodopsin), iClC2, RHO (Rhodopsin), GNAT1(G Protein Subunit Alpha Transducin 1), CNGB1 (Cyclic Nucleotide Gated Channel Subunit Beta 1), APOE (apolipoprotein E), HTRA1 (HtrA serine peptidase 1), or C3 (complement 3). In one embodiment, the expression construct disclosed herein further comprises a post-transcriptional regulatory element. In one embodiment, the post-transcriptional regulatory element is a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). In one embodiment, the post-transcriptional regulatory element comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 11, 38, or 39. In one embodiment, the post-transcriptional regulatory element comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 11, 38, or 39. In one embodiment, the post-transcriptional regulatory element comprises SEQ ID NO: 11, 38, or 39. In one embodiment, the expression construct disclosed herein further comprises a polyadenylation signal (pA). In one embodiment, the polyadenylation signal comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 12. In one embodiment, the pA comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 12. In one embodiment, the pA comprises SEQ ID NO: 12.

[0031] In one aspect, provided is a vector comprising an expression construct disclosed herein comprising a promoter described herein. In one embodiment, the vector is a non-viral vector. In one embodiment, the vector is a viral vector. In some embodiments, the viral vector is selected from the group consisting of adenoviral vector, adeno-associated virus (AAV) vector, and lentiviral vector. In one embodiment, the viral vector is an AAV vector. In some embodiments, the AVV vector comprises inverted terminal repeats (ITRs) derived from AAV2, AAV5, AAV8, or AAV9. In one embodiment, the AAV vector comprises ITRs derived from AAV2. In some embodiments, the AAV vector comprises capsid proteins derived from AAV5 or AAV7m8.

[0032] In one aspect, provided is a cell comprising an expression construct disclosed herein or a vector disclosed herein.

[0033] In one aspect, provided is a pharmaceutical composition comprising (i) an expression construct disclosed herein or a vector disclosed herein and (ii) a pharmaceutically acceptable carrier.

[0034] In one aspect, provided is a method of expressing a transgene in a rod photoreceptor in a subject in need thereof, the method comprising administering to the subject an expression construct disclosed herein, a vector disclosed herein, or a pharmaceutical composition disclosed herein.

[0035] In one aspect, provided is a method of prolonging the survival of cones that otherwise degenerate due to a bystander effect in a subject in need thereof, wherein the method comprises administering to the subject an expression construct disclosed herein, a vector disclosed herein, or a pharmaceutical composition disclosed herein.

[0036] In one aspect, provided is a method of treating an ocular disorder in a subject in need thereof, the method comprising administering to the subject an expression construct disclosed herein, a vector disclosed herein, or a pharmaceutical composition disclosed herein. In one embodiment, the subject has macular degeneration. In one embodiment the ocular disorder is macular degeneration. In one embodiment, the macular degeneration is dry age-related maculardegeneration (AMD), wet age-related AMD, or geographic atrophy (GA). In one embodiment, the subject has a rod dystrophy. In one embodiment the ocular disorder is rod dystrophy. In one embodiment, the subject is a human.BRIEF DESCRIPTION OF THE DRAWINGS

[0037] FIG. 1 provides a schematic of the promoter design approach used in the Examples.

[0038] FIGS. 2A, 2B, and 2C show eGFP fluorescence three weeks post-transduction in retinal organoids transduced with AAV2 / 7m 8 -promoter 3 (TFBS)-eGFP vector. FIG. 2A illustrates a retinal organoid transduced with an AAV2 / 7m8-promoter 3 -eGFP vector. eGFP is expressed across the entirety of the retinal organoid and is not localized to a particular section.FIG. 2B shows a fixed and cryosectioned, retinal organoid that was stained DAPI and eGFP. DAPI expression appeared in the interior of the organoid, while eGFP expression was largely localized to the organoid’s periphery. Scale bar 200 pm and inset 10 pm. FIG. 2C shows the flow cytometry gating strategy with anti-CD73 antibodies to identify rod and cone photoreceptor cells (positive cell populations). Following CD73 gating, photoreceptors were analyzed for expression of eGFP.

[0039] FIGS. 3A and 3B show flow cytometric analysis of eGFP intensity in retinal organoids transduced with AAV2 / 7m8 vectors comprising nucleic acids comprising an eGFP transgene under the control of the indicated promoters. FIG. 3A depicts the median eGFP intensity in CD73 positive / eGFP positive photoreceptor cells. Each dot represents a single transduced retinal organoid. Error bars = SEM (standard error of the mean). FIG. 3B shows the fold change in median eGFP intensity as compared to the RK promoter (dashed line). Median eGFP intensity in eGFP positive / CD73 positive photoreceptors (between 20-61% of CD73 positive cells) was normalized to eGFP expression observed for organoids transduced with a AAV2 / 7m8-RK-eGFP vector. n=5 organoids from n=3 experiments.

[0040] FIGS. 4A, 4B, 4C, 4D, 4E, 4F, 4G, and 4H show the immunohistochemical analysis of eGFP expression in hiPSC retinal organoids transduced with AAV2 / 7m8 vectors comprising nucleic acids comprising an eGFP transgene under the control of the indicated promoters. Retinal organoids were fixed in 4% PFA and cryosectioned at 7 pm. Sections were stained to identify eGFP, L / M opsin, and nuclei (using DAPI). n=2 per group, representative images are shown. Retinal organoids transduced with vectors comprising nucleic acids comprising promoters 3, 4, 5, 6, 7, and 9 showed highly specific rod transduction as indicated by the lack of colocalization with a cone-specific marker (L / M opsin). Note that image acquisition settingswere set higher in the eGFP channel for RK group. Scale bar 40 pm. ROI = region of interest.FIG. 4A Cells transduced with AAV2 / 7m8-RK-eGPF. FIG. 4B Cells transduced with AAV2 / 7m8-promoter 1-eGPF. FIG. 4C Cells transduced with AAV2 / 7m8-promoter 3-eGPF.FIG. 4D Cells transduced with AAV2 / 7m 8 -promoter 4-eGPF FIG. 4E Cells transduced with AAV2 / 7m8-promoter 5-eGPF. FIG. 4F Cells transduced with AAV2 / 7m8-promoter 6-eGPF.FIG. 4G Cells transduced with AAV2 / 7m8-promoter 7-eGPF. FIG. 4H Cells transduced with AAV2 / 7m8-promoter 9-eGPF.

[0041] FIG. 5 illustrates the results of qPCR analysis of eGFP expression in mouse retinal cells transduced with the indicated AAV2 / 5 vectors. Adult WT C57BL / 6 mice were subretinally injected (4 x 109VG / eye) with AAV2 / 5 vectors expressing eGFP under the control of the indicated promoters. Retinas were dissected four weeks post-injection (pi) and analyzed using qPCR. Data displayed are normalized to eGFP expression levels driven by AAV2 / 5-RK-eGFP (dashed line), mean + / - SEM (n = 4 retinas per group).

[0042] FIGS. 6A, 6B, 6C, 6D, 6E, and 6F show immunohistochemical analysis of eGFP expression in mouse eyes. Mice were treated with AAV2 / 5 vectors comprising nucleic acids comprising the eGFP gene under the control of the indicated promoters. Whole eyes were enucleated, fixed in 4 % PF A and embedded in OCT. 18 pm sections were stained with PNA (biotinylated peanut agglutinin) and DAPI. n = 3-4 eyes per group, representative images are shown. Note that image acquisition settings were set higher in the eGFP channel for RK group. ROI = region of interest. Scale bar 50pm. FIG. 6A shows RK promoter activity in cone photoreceptor cells. FIG. 6B shows that the promoter 1 resulted in increased reporter protein expression compared to the RK promoter. FIG.6C shows that promoter 3 resulted in increased reporter protein expression compared to the RK promoter. FIG. 6D shows that promoter 4 exhibited un-specific transduction in the RPE (indicated by the arrowhead). FIG. 6E shows that the promoter 6 resulted in increased reporter protein expression compared to the RK promoter. FIG. 6F shows that the promoter 9 resulted in increased reporter protein expression compared to the RK promoter.

[0043] FIG. 7 illustrates the sequence relationship between promoters 3 and 6-9. Note that the length of the bars is not representative of sequence length.

[0044] FIG. 8 illustrates the median GFP intensity measured in photoreceptors for the RK control promoter and promoters 3, 9, and V12-V16 in human retinal organoids. Human retinal organoids were transduced with AAV2 / 7m8 vectors encoding GFP under control of the indicated promoters. Median GFP intensity in CD73+photoreceptors was determined by flow cytometry. Data are expressed as a fold change relative to expression driven by the RKpromoter. Bars show the mean MFI + / - SEM of n= 8 organoids from two independent transduction experiments). Promoters 3 and 9 increased median GFP intensity in photoreceptor cells by 2-3-fold as compared to the control promoter. Promoters V12-16 increased reporter protein expression 5-9-fold as compared to the control promoter.

[0045] FIG. 9 illustrates that promoter VI 5 promotes rod photoreceptor-specific GFP expression. Cryosections of human retinal organoids were transduced with AAV2 / 7m8-RK-GFP or AAV2 / 7m8-V15-GFP, respectively. GFP expression in the photoreceptor layers was assessed. Promoter VI 5 demonstrated increased GFP expression in photoreceptor cell layer (cell bodies (labeled “photoreceptors”) and outer-segments (labeled “OS”)) as compared to the control promoter. Scale bar = 100 pm and 10 pm (insert).

[0046] FIG. 10 illustrates that promoters V12-V16 do not drive GFP expression in cone photoreceptor cells in human organoid frozen sections. Immunohistochemistry was performed on frozen sections of human organoids transduced with AAV2 / 7m8 vectors comprising nucleic acids encoding GFP under the control of the indicated promoters. L / M opsin labels cone cells. Promoters V12-V16 do not drive GFP expression in cone in cone cells, showing the specificity of V12-V16 for rod cells. * indicates that the channel corresponding to GFP was enhanced to better show GFP fluorescence. Scale bar =25 pm.

[0047] FIG. 11 illustrates eGFP expression level driven by indicated promoters in mouse retina, as assessed by qPCR analysis. Adult WT C57BL / 6 mice were subretinally injected (4 x 109VG / eye) with AAV2 / 5 vectors expressing GFP under control of the indicated promoters. Retinas were dissected four weeks post injection and analyzed using on-target qPCR. Promoters V5 and VI 2- 16 increase GFP expression by up to 28-fold on average (as compared to GFP expression driven by the control promoter). Data shown was normalized to eGFP expression levels driven the RK promoter (dashed line). Mean + / - SEM (n=4-8 per group).

[0048] FIG. 12 demonstrates promoters V5 and VI 2- 16 do not drive GFP expression in cone photoreceptor cells in the mouse retina. Immunohistochemistry was performed on frozen sections of mouse retinas that had been transduced with AAV2 / 5 vectors comprising nucleic acids comprising the GFP transgene under control of the indicated promoters. Arrestin 3 (ARR3) labels cone photoreceptor cells. Promoters V5 and V12-16 do not drive GFP expression in cone in cone cells, showing the specificity of V5 and V12-16 for rod cells. GFP expression driven by the control RK promoter was very weak. As such, to visualize these “dim” expressing cells, the contrast / brightness of images within bottom panel has been increased (*).DETAILED DESCRIPTION

[0049] Provided herein are promoters for the expression of operably linked transgenes in rod photoreceptors. Specifically, promoters disclosed herein can result in high expression levels and / or specific expression of the operatively linked transgenes in mammalian rod photoreceptors, including in murine and / or human rod photoreceptors.

[0050] Gene therapy may serve as a therapeutic treatment for rod disorders by allowing for the expression of a relevant transgene in rod cells. Accordingly, promoters disclosed herein can be used in gene therapy applications to treat rod photoreceptor disorders by allowing both for rod-specific and / or strong expression in rod cells.

[0051] Provided herein are promoters that are rod-specific. Tissue-specific can be used instead of tissue-specific viral capsids frequently used for gene delivery (or can be used in combination with tissue-specific viral capsids). Further, for some gene therapies, it is desirable that gene expression is limited to a specific tissue.

[0052] Provided herein are promoters that more strongly drive gene expression as compared to known reference promoters. Promoters with increased potency allow for the administration of smaller amounts of gene therapy vector, thus decreasing immune responses and associated safety risks. Further, decreasing the amount of a gene therapy vector needed for given therapeutic outcome may also reduce manufacturing costs.

[0053] Rod-Specific Promoters

[0054] Traditionally, promoters are defined as DNA regions where transcription is initiated. A promoter is typically located at the 5' end of the gene (or transcription unit) that the promoter regulates. Promoters are DNA sequences comprising response elements that provide an initial binding site for the RNA transcriptional machinery (including RNA polymerase and transcription factors that recruit the RNA polymerase). Transcription factors bind specific activator and repressor sequences that attach to certain promoters and regulate gene expression. Since the promoter region controls when and where a gene of interest is expressed in an organism, promoters are important elements for regulating the level and specificity of transgene expression, particularly in the context of gene therapy.

[0055] As used herein, a “rod-specific promoter” is a promoter that shows significantly increased, and / or preferential, expression of an operatively linked transgene in rod photoreceptors, as compared to expression in other tissues or cells (such as cone photoreceptors).

[0056] As used herein, “operatively linked” refers to a first molecule joined to a second molecule, wherein the molecules are so arranged that the first molecule affects the function of the second molecule. The two molecules may or may not be part of a single contiguous molecule and may or may not be adjacent. For example, a promoter is operatively linked to a transcribable polynucleotide molecule if the promoter modulates transcription of the transcribable polynucleotide molecule of interest in a cell. Additionally, two portions of a transcription regulatory element are operatively linked to one another if they are joined such that the transcription-activating functionality of one portion is not adversely affected by the presence of the other portion. Two transcription regulatory elements may be operatively linked to one another by way of a linker nucleic acid (e.g., an intervening non-coding nucleic acid) or may be operatively linked to one another with no intervening nucleotides present.

[0057] In some embodiments, the rod-specific promoter comprises a sequence disclosed in Table 3. In some embodiments, the rod-specific promoter comprises a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to a sequence disclosed in Table 3. In some embodiments, the rod-specific promoter comprises any one of SEQ ID NOs: 1-26.

[0058] In one embodiment, the rod-specific promoter comprises a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to ID NO: 1. In one embodiment, the rod-specific promoter comprises a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:2. In one embodiment, the rod-specific promoter comprises a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:3. In one embodiment, the rod-specific promoter comprises a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:4. In one embodiment, the rod-specific promoter comprises a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:5. In one embodiment, the rod-specific promoter comprises a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:6. In one embodiment, the rod-specific promoter comprises a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:8. In one embodiment, the rodspecific promoter comprises a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:9. In one embodiment, the rod-specific promoter comprises a sequence that is at least 80%, at least 85%, at least 90%,at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 10. In one embodiment, the rod-specific promoter comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:22. In one embodiment, the rod-specific promoter comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:23. In one embodiment, the rod-specific promoter comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:24. In one embodiment, the rod-specific promoter comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:25. In one embodiment, the rod-specific promoter comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:26.

[0059] Provided herein is a rod-specific promoter consisting of a sequence comprising (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 18 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17. In one embodiment, the promoter consists of a sequence comprising a nucleic acid sequence comprising (i) SEQ ID NO: 18 and (ii) SEQ ID NO: 17.

[0060] Provided herein is a rod-specific promoter consisting of a sequence comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 18 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17. In one embodiment, the promoter consists of a sequence comprising a nucleic acid sequence comprising from 5’ to 3’ (i) SEQ ID NO: 18 and (ii) SEQ ID NO: 17.

[0061] To illustrate, if, for example, “sequence C comprises sequence A and sequence B from 5’ to 3’,” this means that sequence A is located 5’ with respect to sequence B. In one embodiment, sequence A is located directly 5’ with respect to sequence B, without any intervening sequence. In another embodiment, sequence A is located 5’ with respect to sequence B, but there is an intervening sequence between sequences A and B.

[0062] Provided herein is a rod-specific promoter comprising a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:8. In one embodiment, the promoter comprises a nucleic acid sequence comprising SEQ IDN0:8.

[0063] Provided herein is a rod-specific promoter consisting of a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:8. Provided herein is a rod-specific promoter consisting of SEQ ID N0:8.

[0064] Provided herein is a rod-specific promoter consisting of a sequence comprising (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 18 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17, wherein the promoter comprises no more than about 1050, about 1040, about 1030, about 1020, about 1010, about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, or about 910 nucleotides.

[0065] Provided herein is a rod-specific promoter consisting of a sequence comprising (i) SEQ ID NO: 18 and (ii) SEQ ID NO: 17, wherein the promoter comprises no more than about 1050, about 1040, about 1030, about 1020, about 1010, about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, or about 910 nucleotides.

[0066] Provided herein is a rod-specific promoter consisting of a sequence comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 18 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17, wherein the promoter comprises no more than about 1050, about 1040, about 1030, about 1020, about 1010, about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, or about 910 nucleotides.

[0067] Provided herein is a rod-specific promoter consisting of a sequence comprising from 5’ to 3’ (i) SEQ ID NO: 18 and (ii) SEQ ID NO: 17, wherein the promoter comprises no more than about 1050, about 1040, about 1030, about 1020, about 1010, about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, or about 910 nucleotides.

[0068] Provided herein is a rod-specific promoter consisting of a sequence comprising (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 19 and (ii) a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17. In one embodiment, the promoter consists of a sequence comprising a nucleic acid sequence comprising SEQ ID NO: 19 and SEQ ID NO: 17.

[0069] Provided herein is a rod-specific promoter consisting of a sequence comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 19 and (ii) a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17. In one embodiment, the promoter consists of a sequence comprising a nucleic acid sequence comprising from 5’ to 3’ (i) SEQ ID NO: 19 and (ii) SEQ ID NO: 17.

[0070] Provided herein is a rod-specific promoter comprising a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:9. In one embodiment, the promoter comprises a nucleic acid sequence comprising SEQ IDN0:9.

[0071] Provided herein is a rod-specific promoter consisting of a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:9. Provided herein is a rod-specific promoter consisting of SEQ ID NO:9.

[0072] Provided herein is a rod-specific promoter comprising (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 19 and (ii) a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17, wherein the promoter comprises no more than about 1200, about 1190, about 1180, about 1170, about 1160, about 1150, about 1140, about 1130, about 1120, about 1110, about 1100, about 1090, about 1080,, or about 1073 nucleotides.

[0073] In embodiments, the promoter comprises a nucleic acid sequence comprising SEQ ID NO: 19 and SEQ ID NO: 17, wherein the promoter comprises no more than about 1200, about 1190, about 1180, about 1170, about 1160, about 1150, about 1140, about 1130, about 1120, about 1110, about 1100, about 1090, about 1080, or about 1073 nucleotides.

[0074] Provided herein is a rod-specific promoter comprising (i) optionally the nucleic acid C, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:20, (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:33, (iv) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17, and (v) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:34. In someembodiments, the promoter comprises no more than about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, about 910, about 900, about 890, about 880, about 870, or about 860nucleotides.

[0075] Provided herein is a rod-specific promoter comprising (i) optionally the nucleic acid C, (ii) SEQ ID NO:20, (iii) SEQ ID NO:33, (iv) SEQ ID NO: 17, and (v) SEQ ID NO:34.

[0076] Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) optionally the nucleic acid C, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:20, (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:33, (iv) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17, and (v) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:34.

[0077] Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) optionally the nucleic acid C, (ii) SEQ ID NO:20, (iii) SEQ ID NO:33, (iv) SEQ ID NO: 17, and (v) SEQ ID NO:34.

[0078] Provided herein is a rod-specific promoter comprising (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:35, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:21, and (iii) optionally the nucleic acid sequence CGCGGCCGC. In some embodiments, the promoter comprises no more than about 800, about 790, about 780, about 770, about 760, about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, or about 662 nucleotides.

[0079] Provided herein is a rod-specific promoter comprising (i) SEQ ID NO:35, (ii) SEQ ID NO:21, and (iii) optionally the nucleic acid sequence CGCGGCCGC. Provided herein is a rod-specific promoter comprising (i) SEQ ID NO:35, (ii) SEQ ID NO:21, and (iii) the nucleic acid sequence CGCGGCCGC.

[0080] Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:35, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:21, and (iii) optionally the nucleic acid sequence CGCGGCCGC.

[0081] Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) SEQ ID NO:35, (ii) SEQ ID NO:21, and (iii) optionally the nucleic acid sequence CGCGGCCGC. Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) SEQ ID NO:35, (ii) SEQ ID NO:21, and (iii) the nucleic acid sequence CGCGGCCGC.

[0082] Provided herein is a rod-specific promoter comprising (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:36, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:21, and (iii) optionally the nucleic acid sequence CGCGGCCGC. In some embodiments, the promoter comprises no more than about 1850, about 1840, about 1830, about 1820, about 1810, about 1800, about 1790, about 1780, about 1770, about 1760, about 1750, or about 1713 nucleotides.

[0083] Provided herein is a rod-specific promoter comprising (i) SEQ ID NO:36, (ii) SEQ ID NO:21, and (iii) optionally the nucleic acid sequence CGCGGCCGC. Provided herein is a rod-specific promoter comprising (i) SEQ ID NO:36, (ii) SEQ ID NO:21, and (iii) the nucleic acid sequence CGCGGCCGC.

[0084] Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:36, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:21, and (iii) optionally the nucleic acid sequence CGCGGCCGC.

[0085] Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) SEQ ID NO:36, (ii) SEQ ID NO:21, and (iii) optionally the nucleic acid sequence CGCGGCCGC. Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) SEQ ID NO:36, (ii) SEQ ID NO:21, and (iii) the nucleic acid sequence CGCGGCCGC.

[0086] Provided herein is a rod-specific promoter comprising a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to any one of SEQ ID NO: 10; SEQ ID NON; or SEQ ID NO:3.

[0087] Provided herein is a rod-specific promoter comprising any one of SEQ ID NO: 10; SEQ ID NON; or SEQ ID NO:3.

[0088] Provided herein is a rod-specific promoter consisting of a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to any one of SEQ ID NO: 10, SEQ ID NON, or SEQ ID NON.

[0089] Provided herein is a rod-specific promoter consisting of any one of SEQ ID NO: 10, SEQ ID NON, or SEQ ID NON.

[0090] Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:20 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:6. Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:20 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:6. Further provided herein is a rod-specific promoter comprising SEQ ID NO:20 and SEQ ID NO:6.

[0091] Provided herein is a rod-specific promoter comprising (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:20 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:6. Provided herein is a rod-specific promoter comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:20 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:6. Further provided herein is a rod-specific promoter comprising SEQ ID NO:20 and SEQ IDNO:6.

[0092] Provided herein is a rod-specific promoter that is derived from SEQ ID NO: 13. Provided herein is a rod-specific promoter that is no longer than about 2000, about 1990, about 1980, about 1970, about 1960, about 1950, about 1940, about 1930, about 1920, about 1910, about 1900, about 1890, about 1880, about 1870, about 1860, about 1850, about 1840, about 1830, about 1820, about 1810, about 1800, about 1790, about 1780, about 1770, about 1760, about 1750, about 1740, about 1730, about 1720, about 1710, about 1700, about 1690, about 1680, about 1670, about 1660, about 1650, about 1640, about 1630, about 1620, about 1610, about 1600, about 1590, about 1580, about 1570, about 1560, about 1550, about 1540, about 1530, about 1520, about 1510, about 1500, about 1490, about 1480, about 1470, about 1460, about 1450, about 1440, about 1430, about 1420, about 1410, about 1400, about 1390, about 1380, about 1370, about 1360, about 1350, about 1340, about 1330, about 1320, about 1310, about 1300, about 1290, about 1280, about 1270, about 1260, about 1250, about 1240, about1230, about 1220, about 1210, about 1200, about 1190, about 1180, about 1170, about 1160, about 1150, about 1140, about 1130, about 1120, about 1110, about 1100, about 1090, about 1080, about 1070, about 1060, about 1050, about 1040, about 1030, about 1020, about 1010, about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, about 910, about 900, about 890, about 880, about 870, about 860, about 850, about 840, about 830, about 820, about 810, about 800, about 790, about 780, about 770, about 760, about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about 640, about 630, about 620, about 610, about 600, about 590, about 580, about 570, or about 560 nucleotides and that (a) comprises a fragment of contiguous nucleotides from SEQ ID NO: 13 or (b) a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to the fragment of (a).

[0093] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 1, wherein the promoter comprises no more than about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about 640, about 630, about 620, about 610, about 600, about 590, about 580, about 570, or about 560 contiguous nucleotides from SEQ ID NO: 13.

[0094] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 1, wherein the promoter comprises no more than about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about 640, about 630, about 620, about 610, about 600, about 590, about 580, about 570, or about 560 nucleotides in length.

[0095] Provided herein rod-specific promoter that consists of SEQ ID NO: 1.

[0096] Provided herein is a rod-specific promoter that is derived from SEQ ID NO: 15. Provided herein is a rod-specific promoter that is no longer than about 2000, about 1990, about 1980, about 1970, about 1960, about 1950, about 1940, about 1930, about 1920, about 1910, about 1900, about 1890, about 1880, about 1870, about 1860, about 1850, about 1840, about 1830, about 1820, about 1810, about 1800, about 1790, about 1780, about 1770, about 1760, about 1750, about 1740, about 1730, about 1720, about 1710, about 1700, about 1690, about 1680, about 1670, about 1660, about 1650, about 1640, about 1630, about 1620, about 1610, about 1600, about 1590, about 1580, about 1570, about 1560, about 1550, about 1540, about 1530, about 1520, about 1510, about 1500, about 1490, about 1480, about 1470, about 1460, about 1450, about 1440, about 1430, about 1420, about 1410, about 1400, about 1390, about1380, about 1370, about 1360, about 1350, about 1340, about 1330, about 1320, about 1310, about 1300, about 1290, about 1280, about 1270, about 1260, about 1250, about 1240, about 1230, about 1220, about 1210, about 1200, about 1190, about 1180, about 1170, about 1160, about 1150, about 1140, about 1130, about 1120, about 1110, about 1100, about 1090, about 1080, about 1070, about 1060, about 1050, about 1040, about 1030, about 1020, about 1010, about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, about 910, about 900, about 890, about 880, about 870, about 860, about 850, about 840, about 830, about 820, about 810, about 800, about 790, about 780, about 770, about 760, about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about 640, about 630, about 620, about 610, about 600, about 590, about 580, about 570, about 560, about 550, about 540, about 530, about 520, about 510, about 500, about 490, about 480, about 470, about 460, about 450, about 440, about 430, about 420, about 410, about 400, about 390, about 380, about 370, about 360, about 350, about 340, about 330, about 320, about 310, about 300, about 290, about 280, about 270, about 260, about 250, about 240, about 230, about 220 or about 210 nucleotides and that (a) comprises a fragment of contiguous nucleotides from SEQ ID NO: 15 or (b) a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to the fragment of (a).

[0097] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 5, wherein the promoter comprises no more than about 400, about 390, about 380, about 370, about 360, about 350, about 340, about 330, about 320, about 310, about 300, about 290, about 280, about 270, about 260, about 250, about 240, about 230, about 220, or about 210 contiguous nucleotides from SEQ ID NO: 15.

[0098] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 5, wherein the promoter comprises no more than about 400, about 390, about 380, about 370, about 360, about 350, about 340, about 330, about 320, about 310, about 300, about 290, about 280, about 270, about 260, about 250, about 240, about 230, about 220, or about 210 nucleotides in length.

[0099] Provided herein rod-specific promoter that consists of SEQ ID NO:5.

[0100] Provided herein is a rod-specific promoter that is derived from one or more fragments of SEQ ID NO: 16.

[0101] Provided herein is a rod-specific promoter comprising one or more components, wherein each component is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to a fragment of SEQ ID NO: 16.

[0102] Provided herein is a rod-specific promoter comprising one or more fragments of SEQ ID NO: 16.

[0103] Provided herein is a rod-specific promoter that is no longer than about 2000, about 1990, about 1980, about 1970, about 1960, about 1950, about 1940, about 1930, about 1920, about 1910, about 1900, about 1890, about 1880, about 1870, about 1860, about 1850, about 1840, about 1830, about 1820, about 1810, about 1800, about 1790, about 1780, about 1770, about 1760, about 1750, about 1740, about 1730, about 1720, about 1710, about 1700, about 1690, about 1680, about 1670, about 1660, about 1650, about 1640, about 1630, about 1620, about 1610, about 1600, about 1590, about 1580, about 1570, about 1560, about 1550, about 1540, about 1530, about 1520, about 1510, about 1500, about 1490, about 1480, about 1470, about 1460, about 1450, about 1440, about 1430, about 1420, about 1410, about 1400, about 1390, about 1380, about 1370, about 1360, about 1350, about 1340, about 1330, about 1320, about 1310, about 1300, about 1290, about 1280, about 1270, about 1260, about 1250, about 1240, about 1230, about 1220, about 1210, about 1200, about 1190, about 1180, about 1170, about 1160, about 1150, about 1140, about 1130, about 1120, about 1110, about 1100, about 1090, about 1080, about 1070, about 1060, about 1050, about 1040, about 1030, about 1020, about 1010, about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, about 910, about 900, about 890, about 880, about 870, about 860, about 850, about 840, about 830, about 820, about 810, about 800, about 790, about 780, about 770, about 760, about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about 640, about 630, about 620, about 610, or about 600 nucleotides and that (a) comprises a fragment of contiguous nucleotides from SEQ ID NO: 16 or (b) a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to the fragment of (a).

[0104] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 6, wherein the promoter comprises no more than about 900, about 890, about 880, about 870, about 860, about 850, about 840, about 830, about 820, about 810, about 800, about 790, about 780, about 770, about 760, about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about640, about 630, about 620, about 610, or about 600 contiguous nucleotides from SEQ ID NO:16.

[0105] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 6, wherein the promoter comprises no more than about 900, about 890, about 880, about 870, about 860, about 850, about 840, about 830, about 820, about 810, about 800, about 790, about 780, about 770, about 760, about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about 640, about 630, about 620, about 610, or about 600 nucleotides in length.

[0106] Provided herein is a rod-specific promoter that consists of a sequence comprising (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:33, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17, and (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:34. In some embodiments, the promoter comprises no more than about 900, about 890, about 880, about 870, about 860, about 850, about 840, about 830, about 820, about 810, about 800, about 790, about 780, about 770, about 760, about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about 640, about 630, about 620, about 610, or about 600 nucleotides in length.

[0107] Provided herein is a rod-specific promoter that consists of a sequence comprising (i) SEQ IDNO:33, (ii) SEQ ID NO: 17, and (iii) SEQ ID NO:34.

[0108] Provided herein is a rod-specific promoter that consists of a sequence comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:33, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 17, and (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ IDNO:34.

[0109] Provided herein is a rod-specific promoter that consists of a sequence comprising from 5’ to 3’ (i) SEQ ID NO:33, (ii) SEQ ID NO: 17, and (iii) SEQ IDNO:34.

[0110] Provided herein is a rod-specific promoter that consists of SEQ ID NO:6.[OHl] Provided herein is a rod-specific promoter that is derived from SEQ ID NO: 14. Provided herein is a rod-specific promoter that is no longer than about 2000, about 1990, about1980, about 1970, about 1960, about 1950, about 1940, about 1930, about 1920, about 1910, about 1900, about 1890, about 1880, about 1870, about 1860, about 1850, about 1840, about 1830, about 1820, about 1810, about 1800, about 1790, about 1780, about 1770, about 1760, about 1750, about 1740, about 1730, about 1720, about 1710, about 1700, about 1690, about 1680, about 1670, about 1660, about 1650, about 1640, about 1630, about 1620, about 1610, about 1600, about 1590, about 1580, about 1570, about 1560, about 1550, about 1540, about 1530, about 1520, about 1510, about 1500, about 1490, about 1480, about 1470, about 1460, about 1450, about 1440, about 1430, about 1420, about 1410, about 1400, about 1390, about 1380, about 1370, about 1360, about 1350, about 1340, about 1330, about 1320, about 1310, about 1300, about 1290, about 1280, about 1270, about 1260, about 1250, about 1240, about 1230, about 1220, about 1210, about 1200, about 1190, about 1180, about 1170, about 1160, about 1150, about 1140, about 1130, about 1120, about 1110, about 1100, about 1090, about 1080, about 1070, about 1060, about 1050, about 1040, about 1030, about 1020, about 1010, about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, about 910, about 900, about 890, about 880, about 870, about 860, about 850, about 840, about 830, about 820, about 810, about 800, about 790, about 780, about 770, about 760, about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about 640, about 630, about 620, about 610, about 600, about 590, about 580, about 570, about 560, about 550, about 540, about 530, about 520, about 510, about 500, about 490, about 480, about 470, about 460, about 450, about 440, about 430, about 420, about 410, about 400, about 390, about 380, about 370, about 360, about 350, about 340, about 330, about 320, about 310, about 300, about 290, about 280, or about 270 nucleotides and that (a) comprises a fragment of contiguous nucleotides from SEQ ID NO: 14 or (b) a sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to the fragment of (a).

[0112] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:2, wherein the promoter comprises no more than about 400, about 390, about 380, about 370, about 360, about 350, about 340, about 330, about 320, about 310, about 300, about 290, about 280, or about 270 contiguous nucleotides from SEQ ID NO: 14.

[0113] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:2, wherein the promoter comprises no more than about 400,about 390, about 380, about 370, about 360, about 350, about 340, about 330, about 320, about 310, about 300, about 290, about 280, or about 270 nucleotides in length. Provided herein is a rod-specific promoter that consists of SEQ ID NO:2.

[0114] Provided herein is a rod-specific promoter that consists of a sequence comprising (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:37 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:21. In embodiments, the promoter comprises no more than about 400, about 390, about 380, about 370, about 360, about 350, about 340, about 330, about 320, about 310, about 300, about 290, about 280, or about 270 nucleotides in length.

[0115] Provided herein is a rod-specific promoter that consists of a sequence comprising (i) SEQ ID NO:37 and (ii) SEQ ID NO:21.

[0116] Provided herein is a rod-specific promoter that consists of a sequence comprising from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:37 and (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:21.

[0117] Provided herein is a rod-specific promoter that consists of a sequence comprising from 5’ to 3’ (i) SEQ ID NO:37 and (ii) SEQ IDN0:21.

[0118] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:21. Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:21, wherein the promoter is no longer than about 2000, about 1990, about 1980, about 1970, about 1960, about 1950, about 1940, about 1930, about 1920, about 1910, about 1900, about 1890, about 1880, about 1870, about 1860, about 1850, about 1840, about 1830, about 1820, about 1810, about 1800, about 1790, about 1780, about 1770, about 1760, about 1750, about 1740, about 1730, about 1720, about 1710, about 1700, about 1690, about 1680, about 1670, about 1660, about 1650, about 1640, about 1630, about 1620, about 1610, about 1600, about 1590, about 1580, about 1570, about 1560, about 1550, about 1540, about 1530, about 1520, about 1510, about 1500, about 1490, about 1480, about 1470, about 1460, about 1450, about 1440, about 1430, about 1420, about 1410, about 1400, about 1390, about 1380, about 1370, about 1360, about 1350, about 1340, about 1330, about 1320, about 1310, about 1300, about 1290, about 1280, about 1270,about 1260, about 1250, about 1240, about 1230, about 1220, about 1210, about 1200, about 1190, about 1180, about 1170, about 1160, about 1150, about 1140, about 1130, about 1120, about 1110, about 1100, about 1090, about 1080, about 1070, about 1060, about 1050, about 1040, about 1030, about 1020, about 1010, about 1000, about 990, about 980, about 970, about 960, about 950, about 940, about 930, about 920, about 910, about 900, about 890, about 880, about 870, about 860, about 850, about 840, about 830, about 820, about 810, about 800, about 790, about 780, about 770, about 760, about 750, about 740, about 730, about 720, about 710, about 700, about 690, about 680, about 670, about 660, about 650, about 640, about 630, about 620, about 610, about 600, about 590, about 580, about 570, about 560, about 550, about 540, about 530, about 520, about 510, about 500, about 490, about 480, about 470, about 460, about 450, about 440, about 430, about 420, about 410, about 400, about 390, about 380, about 370, about 360, about 350, about 340, about 330, about 320, about 310, about 300, about 290, about 280, about 270, about 260, or about 250 nucleotides in length.

[0119] Provided herein is a rod-specific promoter that comprises (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 18, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 27, (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, and (iv) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 29. In embodiments, the promoter comprises no more than about 1600, about 1590, about 1580, about 1570, about 1560, about 1550, about 1540, about 1530, about 1520, about 1510, about 1500, about 1490, about 1480, or about 1474 nucleotides.

[0120] Provided herein is a rod-specific promoter that comprises (i) SEQ ID NO: 18, (ii) SEQ ID NO: 27, (iii) SEQ ID NO: 28, and (iv) SEQ ID NO: 29.

[0121] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 18, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 27, (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, and (iv) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 29. In embodiments, the promotercomprises no more than about 1600, about 1590, about 1580, about 1570, about 1560, about 1550, about 1540, about 1530, about 1520, about 1510, about 1500, about 1490, about 1480, or about 1474 nucleotides.

[0122] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) SEQ ID NO: 18, (ii) SEQ ID NO: 27, (iii) SEQ ID NO: 28, and (iv) SEQ ID NO: 29.

[0123] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:22. In one embodiment, the promoter comprises SEQ ID NO:22.

[0124] Provided herein is a rod-specific promoter that comprises (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 19, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 27 (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, and (iv) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 29. In embodiments, the promoter comprises no more than about 1750, about 1740, about 1730, about 1720, about 1710, about 1700, about 1690, about 1680, about 1670, about 1660, about 1650, about 1640, or about 1637 nucleotides.

[0125] Provided herein is a rod-specific promoter that comprises (i) SEQ ID NO: 19, (ii) SEQ ID NO: 27 (iii) SEQ ID NO: 28, and (iv) SEQ ID NO: 29.

[0126] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 19, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 27 (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, and (iv) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 29. In embodiments, the promoter comprises no more than about 1750, about 1740, about 1730, about 1720, about 1710, about 1700, about 1690, about 1680, about 1670, about 1660, about 1650, about 1640, or about 1637 nucleotides.

[0127] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) SEQ ID NO: 19, (ii) SEQ ID NO: 27 (iii) SEQ ID NO: 28, and (iv) SEQ ID NO: 29.

[0128] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:23. In one embodiment, the promoter comprises SEQ ID NO:23.

[0129] Provided herein is a rod-specific promoter that comprises (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 19, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 27, and (iv) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 30. In embodiments, the promoter comprises no more than about 1750, about 1740, about 1730, about 1720, about 1710, about 1700, about 1690, about 1680, about 1670, about 1660, about 1650, about 1640, or about 1636 nucleotides.

[0130] Provided herein is a rod-specific promoter that comprises (i) SEQ ID NO: 19, (ii) SEQ ID NO: 28, (iii) SEQ ID NO: 27, and (iv) SEQ ID NO: 30.

[0131] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 19, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 27, and (iv) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 30. In embodiments, the promoter comprises no more than about 1750, about 1740, about 1730, about 1720, about 1710, about 1700, about 1690, about 1680, about 1670, about 1660, about 1650, about 1640, or about 1636 nucleotides.

[0132] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) SEQ ID NO: 19, (ii) SEQ ID NO: 28, (iii) SEQ ID NO: 27, and (iv) SEQ ID NO: 30.

[0133] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:24. In one embodiment, the promoter comprises SEQ ID NO:24.

[0134] Provided herein is a rod-specific promoter that comprises (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 19, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, and (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 29. In embodiments, the promoter comprises no more than about 1350, about 1340, about 1330, about 1320, about 1310, about 1300, about 1290, about 1280, about 1270, about 1260, about 1250, about 1240, about 1230, about 1220, or about 1213 nucleotides.

[0135] Provided herein is a rod-specific promoter that comprises (i) SEQ ID NO: 19, (ii) SEQ ID NO: 27 (iii) SEQ ID NO: 28, and (iv) SEQ ID NO: 29.

[0136] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 19, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, and (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 29.

[0137] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) SEQ ID NO: 19, (ii) SEQ ID NO: 27 (iii) SEQ ID NO: 28, and (iv) SEQ ID NO: 29.

[0138] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:25. In one embodiment, the promoter comprises SEQ ID NO:25.

[0139] Provided herein is a rod-specific promoter that comprises (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 31, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, and (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 32. In embodiments, the promoter comprises no more than about 1600, about 1590, about 1580, about 1570, about 1560, about 1550, about 1540, about 1530, about 1520, about 1510, about 1500, about 1490, about 1480, or about 1470 nucleotides.

[0140] Provided herein is a rod-specific promoter that comprises (i) SEQ ID NO: 31, (ii) SEQ ID NO: 28, and (iii) SEQ ID NO: 32.

[0141] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 31, (ii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 28, and (iii) a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 32. In embodiments, the promoter comprises no more than about 1600, about 1590, about 1580, about 1570, about 1560, about 1550, about 1540, about 1530, about 1520, about 1510, about 1500, about 1490, about 1480, or about 1470 nucleotides.

[0142] Provided herein is a rod-specific promoter that comprises from 5’ to 3’ (i) SEQ ID NO: 31, (ii) SEQ ID NO: 28, and (iii) SEQ ID NO: 32.

[0143] Provided herein is a rod-specific promoter that comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:26. In one embodiment, the promoter comprises SEQ ID NO:26.

[0144] In embodiments, the rod-specific promoter consists of a nucleic acid sequence that is at least 80% identical to any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NOs:22-26. In embodiments, the rod-specific promoter consists of a nucleic acid sequence that is at least 90% identical to any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NOs:22-26. In embodiments, the rod-specific promoter consists of any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NOs: 22-26.

[0145] Sequence identity may be calculated using any suitable algorithm. For example, the PILEUP and BLAST algorithms can be used to calculate identity or line up sequences (such as identifying equivalent or corresponding sequences (typically on their default settings), for example as described in Altschul, A protein alignment scoring system sensitive at all evolutionary distances. J Mol Evol. 1993 Mar;36(3):290-300; Altschul et al., Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403-10. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (ncbi.nlm.nih.gov / ). This algorithm involves first identifying high scoring sequence pair (HSPs) by identifying short words of length W in the query sequence that either match or satisfy some positive- valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold. These initial neighborhood word hits act as seeds for initiating searches to find HSPs containing them. Theword hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Extensions for the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment. The BLAST program uses as defaults a word length (W) of 11, the BLOSUM62 scoring matrix alignments (B) of 50, expectation (E) of 10, M=5, N=4, and a comparison of both strands. See Henikoff & Henikoff, Amino acid substitution matrices from protein blocks, Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10915-9.

[0146] The BLAST algorithm performs a statistical analysis of the similarity between two sequences. See, e.g., Karlin & Altschul, Applications and statistics for multiple high-scoring segments in molecular sequences., Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5873-7. One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two polynucleotide or amino acid sequences would occur by chance. For example, a sequence is considered similar to another sequence if the smallest sum probability in comparison of the first sequence to the second sequence is less than about 1, preferably less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.

[0147] Alternatively, the UWGCG Package provides the BESTFIT program which can be used to calculate identity (for example used on its default settings). See Devereux et al., A comprehensive set of sequence analysis programs for the VAX, Nucleic Acids Res. 1984 Jan 11;12(1 Pt l):387-95.

[0148] Also provided herein are promoters that are variants of any of the promoters disclosed herein. Contemplated herein are promoter variants that include sequence in addition (e.g., added at either end or inserted within the promoter sequence) to the elements of a promoter disclosed herein. Likewise, provided are promoter variants in which certain sequence has been removed from promoters disclosed herein (e.g., as a terminal or internal deletion). Provided herein are promoter variants in which at least 10 consecutive nucleotides, at least 15 consecutive nucleotides, at least 20 consecutive nucleotides, at least 25 consecutive nucleotides, at least 30 consecutive nucleotides, at least 35 consecutive nucleotides, at least 40 consecutive nucleotides, at least 45 consecutive nucleotides, or at least 50 consecutive nucleotides have been removed from either the 5’ or the 3’ end of a promoter disclosed herein. Provided herein are promoter variants in which no more than 10 consecutive nucleotides, nomore than 15 consecutive nucleotides, no more than 20 consecutive nucleotides, no more than 25 consecutive nucleotides, no more than 30 consecutive nucleotides, no more than 35 consecutive nucleotides, no more than 40 consecutive nucleotides, no more than 45 consecutive nucleotides, or no more than 50 consecutive nucleotides have been removed from either the 5’ or the 3’ end of a promoter disclosed herein. Provided herein are promoter variants in which 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides have been added to the 5’ and / or the 3’ end of a promoter disclosed herein.

[0149] Transgenes

[0150] In some embodiments, the rod-specific promoters disclosed drive expression of a transgene of interest. In one embodiment, the transgene comprises a protein-coding sequence (e.g., encoding a therapeutic protein).

[0151] In one embodiment, the transgene comprises a protein-coding sequence that encodes a light-sensitive or light-gated G-coupled membrane protein, ion channel, ion pump, or ion transporter. Preferred light-sensitive proteins include ArchT, Jaws (cruxhalorhodopsin), and iQC2. Alternatively, the protein may itself not be directly light-sensitive, but may indirectly modulate endogenous light-sensitive signaling in a rod photoreceptor cell. Examples of such proteins are members of the RGS9 complex, notably R9AP (also known as Rgs9-anchor protein or RGS9BP), and GNB5. In embodiments, the transgene comprises a protein-coding sequence that encodes any other gene product that increases the speed of the endogenous rod signaling mechanism.

[0152] Examples of additional transgenes that can be used with the compositions and methods disclosed herein include, but are not limited to Rhodopsin, GNAT1 (G Protein Subunit Alpha Transducin 1), CNGB1, APOE, HTRA1, or C3.

[0153] Expression Constructs

[0154] The term “expression construct” refers to a recombinant polynucleotide construct that includes a nucleic acid encoding an RNA that is capable of being transcribed in a cell. Methods for constructing expression constructs and plasmids are known in the art.

[0155] Provided herein are expression constructs comprising a rod-specific promoter disclosed herein. Provided herein are expression constructs comprising a rod-specific promoter disclosed herein operatively linked to a transgene, including a transgene disclosed herein.

[0156] The expression constructs disclosed herein may comprise additional regulatory elements, including, but not limited to, enhancers, translation initiation signals, introns, and / orsplicing enhancers. In embodiments, the expression constructs disclosed herein comprise an internal ribosome entry site (IRES). An IRES sequence may be used to produce more than one polypeptide from a single gene transcript. An IRES (or other suitable sequence) is used to produce a protein that contains more than one polypeptide chain or to express two different proteins from or within the same cell.

[0157] The expression constructs disclosed herein may include regulatory elements such as woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) and a polyadenylation signal (pA).

[0158] WPRE sequences are known in the art. In embodiments, the expression construct comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 11 (WPRE6). In embodiments, the expression construct comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:38 (WPRE3). In embodiments, the expression construct comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO:39 (WPRE250). See Table 4. In one embodiment, the expression construct comprises SEQ ID NO: 11, 38, or 39. In some embodiments, the expression construct as described comprises any type of WPRE. Examples of WPRE include, but are not limited to WPRE250 (SQ ID NO:39), WPRE3 (SEQ ID NO:38), and WPRE6 (SEQ ID NO: 11). In some embodiments, a WPRE sequence described in Zanta-Boussif et al., Validation of a mutated PRE sequence allowing high and sustained transgene expression while abrogating WHV-X protein synthesis: application to the gene therapy of WAS. Gene Ther.2009 May;16(5):605-19 is used.

[0159] Polyadenylation sequences are known in the art. In one embodiment, the expression construct comprises BGHpA (bovine growth hormone poly A). In embodiments, the expression construct comprises a nucleic acid sequence that is at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% identical to SEQ ID NO: 12. See Table 4.In one embodiment, the expression construct comprises SEQ ID NO: 12. In one embodiment, provided is a polyadenylation site disclosed in Herrmann et al., PolyASite 2.0: a consolidated atlas of polyadenylation sites from 3' end sequencing. Nucleic Acids Res. 2020 Jan 8;48(D1):D174-D179.

[0160] Vectors

[0161] “Vector,” as used herein, refers to a vehicle that comprises a polynucleotide (e.g., that comprises an expression construct disclosed herein) to be delivered into a host cell, either in vitro, ex vivo or in vivo. Non-limiting examples of vectors include a recombinant plasmid, yeast artificial chromosome (YAC), mini chromosome, DNA mini-circle, or a virus (including virus derived sequences). A vector may also refer to a virion comprising a nucleic acid (e.g., a nucleic acid comprising an expression construct disclosed herein) to be delivered into a host cell, either in vitro, ex vivo or in vivo. In one embodiment, a vector refers to a virion comprising a recombinant viral genome, wherein the viral genome comprises one or more ITRs and a transgene. In one embodiment, a vector refers to a virion comprising a recombinant viral genome, wherein the viral genome comprises one or more ITRs and an expression construct comprising a transgene. In one embodiment, the recombinant vector is a viral vector or a combination of multiple viral vectors. In one aspect, provided is a vector comprising any of the expression constructs disclosed herein.

[0162] For a review of viral and non-viral delivery systems for gene delivery see Nayerossadat et al., Viral and nonviral delivery systems for gene delivery, Adv Biomed Res.2012;l:27.

[0163] Viral Vectors

[0164] Viral vectors for the expression of a target gene in a target cell, tissue, or organism are known in the art and include, for example, an AAV vector, adenovirus vector, lentivirus vector, retrovirus vector, poxvirus vector, baculovirus vector, herpes simplex virus vector, vaccinia virus vector, or a synthetic virus vector (e.g., a chimeric virus, mosaic virus, or pseudotyped virus, and / or a virus that contains a foreign protein, synthetic polymer, nanoparticle, or small molecule).

[0165] AA V vectors

[0166] Adeno-associated viruses (AAV) are small, single-stranded DNA viruses which require helper virus to facilitate efficient replication. The 4.7 kb genome of AAV is characterized by two inverted terminal repeats (ITRs) and two open reading frames which encode the Rep proteins and Cap proteins, respectively. The Rep reading frame encodes four proteins with molecular weights of 78 kD, 68 kD, 52 kD, and 40 kD, respectively. These proteins function mainly in regulating AAV replication and rescue and integration of the AAV into a host cell’s chromosomes. The Cap reading frame encodes three structural proteins with molecular weights of 85 kD (VP 1), 72 kD (VP2), and 61 kD (VP3), which form the virion capsid. More than 80% of total proteins in AAV virion comprise VP3. Flanking the rep andcap open reading frames at the 5' and 3' ends are about 145 bp long ITRs. The two ITRs are the only cis elements important for AAV replication, rescue, packaging, and integration of the AAV genome. The entire rep and cap domains can be excised and replaced with a therapeutic or reporter transgene.

[0167] Recombinant adeno-associated virus “rAAV” vectors include any vector derived from any adeno-associated virus serotype. rAAV vectors can have one or more of the AAV wild-type genes deleted in whole or in part, preferably the Rep and / or Cap genes, but retain functional flanking ITR sequences.

[0168] In some embodiments, the viral vector is an rAAV virion that comprises an rAAV genome and one or more capsid proteins. In some embodiments, the rAAV genome comprises an expression construct disclosed herein.

[0169] In some embodiments, the viral vector disclosed herein comprises a nucleic acid comprising an AAV 5 ' ITR and 3 ' ITR located 5 ' and 3 ' of the transgene, respectively. However, in certain embodiments, it may be desirable for the nucleic acid to contain the 5' ITR and 3' ITR sequences arranged in tandem, e.g., 5' to 3' or a head-to-tail, or in another alternative configuration. In still other embodiments, it may be desirable for the nucleic acid to contain multiple copies of the ITRs or to have 5' ITRs (or conversely, 3' ITRs) located both 5' and 3' to the transgene. The ITRs sequences may be located immediately upstream and / or downstream of the heterologous molecule, or there may be intervening sequences. The ITRs need not be the wild-type nucleotide sequences, and may be altered (e.g., by the insertion, deletion, or substitution of nucleotides) so long as the sequences provide for functional rescue, replication, and packaging. The ITRs may be selected from AAV2, or from among the other AAV serotypes, e.g., as described herein.

[0170] In some embodiments, provided is a vector comprising a nucleic acid sequence comprising (i) an expression construct disclosed herein and (ii) one or more ITRs. In one embodiment, the nucleic acid sequence comprises a 5' ITR and a 3' ITR. In one embodiment, the 5' ITR and the 3' ITR are derived from AAV serotype AAV2. In one embodiment, the 5' ITR and the 3' ITR are derived from AAV serotype AAV5.

[0171] In some embodiments, the viral vector is an AAV vector, such as an AAV1 (z.e., an AAV containing AAV1 ITRs and AAV1 capsid proteins), AAV2 (z.e., an AAV containing AAV2 ITRs and AAV2 capsid proteins), AAV3 (i.e., an AAV containing AAV3 ITRs and AAV3 capsid proteins), AAV4 (z.e., an AAV containing AAV4 ITRs and AAV4 capsid proteins), AAV5 (z.e., an AAV containing AAV5 ITRs and AAV5 capsid proteins), AAV6 (z.e., an AAV containing AAV6 ITRs and AAV6 capsid proteins), AAV7 (z.e., an AAVcontaining AAV7 ITRs and AAV7 capsid proteins), AAV8 (z.e., an AAV containing AAV8 ITRs and AAV8 capsid proteins), AAV9 (z.e., an AAV containing AAV9 ITRs and AAV9 capsid proteins), AAVrh74 (z.e., an AAV containing AAVrh74 ITRs and AAVrh74 capsid proteins), AAVrh.8 (z.e., an AAV containing AAVrh.8 ITRs and AAVrh.8 capsid proteins), or AAVrh.10 (z.e., an AAV containing AAVrh.10 ITRs and AAVrh.10 capsid proteins).

[0172] In some embodiments, the viral vector is a pseudotyped AAV vector, containing ITRs from one AAV serotype and capsid proteins from a different AAV serotype. In some embodiments, the pseudotyped AAV is AAV2 / 9 (z.e., an AAV containing AAV2 derived ITRs and AAV9 capsid proteins). In some embodiments, the pseudotyped AAV is AAV2 / 10 (z.e., an AAV containing AAV2 derived ITRs and AAV10 capsid proteins). In some embodiments, the pseudotyped AAV is AAV2 / 7m8 (z.e., an AAV containing AAV2 derived ITRs and AAV7m8 capsid proteins).

[0173] In some embodiments, the AAV vector contains a recombinant capsid protein, such as a capsid protein containing a chimera of one or more of capsid proteins from AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAVrh74, AAVrh.8, or AAVrh.10. In embodiments, the capsid protein is a variant AAV capsid protein. In one embodiment, the capsid protein is AAV2 variant rAAV2-retro (SEQ ID NO:44 from WO 2017 / 218842, incorporated herein by reference in its entirety). In embodiments, the capsid protein comprises a sequence that is at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a capsid sequence disclosed in WO2017 / 218842.

[0174] Other viral vectors

[0175] Other viral vectors include adenoviral (AV) vectors, for example, those based on human adenovirus type 2 and human adenovirus type 5 that have been made replication defective through deletions in the El and E3 regions. The transcriptional cassette can be inserted into the El region, yielding a recombinant El / E3-deleted AV vector. Adenoviral vectors also include helper-dependent high-capacity adenoviral vectors (also known as high-capacity, “gutless” or “gutted” vectors), which do not contain viral coding sequences. These vectors contain the cis-acting elements needed for viral DNA replication and packaging, mainly the ITR and the packaging signal (CY). These helper-dependent AV vector genomes have the potential to carry from a few hundred base pairs up to approximately 36 kb of foreign DNA.

[0176] Alternatively, other systems such as lentiviral vectors can be used. Lentiviral-based systems can transduce nondividing as well as dividing cells. Lentiviral vectors are derived fromthe human immunodeficiency virus and, like that virus, integrate into the host genome providing the potential for very long-term gene expression.

[0177] Polynucleotides, including plasmids, YACs, minichromosomes and minicircles, comprising an expression construct disclosed herein can also be introduced into a cell or organism by nonviral vector systems using, for example, cationic lipids, polymers, or both as carriers. Conjugated poly-L-lysine (PLL) polymer and polyethylenimine (PEI) polymer systems can also be used to deliver the vector to cells. Other methods for delivering the vector to cells includes hydrodynamic injection and electroporation and use of ultrasound, both for cell culture and for organisms. For a review of viral and non-viral delivery systems for gene delivery see Nayerossadat et al., Viral and nonviral delivery systems for gene delivery, Adv Biomed Res. 2012; 1:27.

[0178] Cells

[0179] In some embodiments, the expression constructs and vectors disclosed herein are used to deliver a nucleic acid comprising a rod-specific promoter operatively linked to a transgene to a cell. Provided herein are cells comprising a promoter, an expression constructs, or a vector disclosed herein. In one embodiment, the cells are photoreceptor cells, in particular rod photoreceptor cells. The cell may be an isolated cell.

[0180] In some embodiments, the cell is a mammalian cell, e.g., a mouse, cow, pig, camel, llama, horse, goat, rabbit, sheep, hamsters, guinea pig, cat, dog cell. In one embodiment, the cell a non-human primate cell. The non-human primate may be, for example, a monkey, such as a cynomolgus monkey, chimpanzee, etc. In one embodiment, the cell is a human cell.

[0181] Pharmaceutical Compositions

[0182] Provided herein are pharmaceutical compositions comprising (i) an expression construct or vector disclosed herein and (ii) a pharmaceutically acceptable excipient. The expression construct or vector disclosed herein is preferably assessed for contamination by conventional methods and formulated into a pharmaceutical composition suitable for storage and / or administration to a patient.

[0183] Formulations of the expression constructs or vectors disclosed herein disclosed herein involve the use of a pharmaceutically and / or physiologically acceptable vehicle or carrier, particularly one suitable for subretinal injection, such as buffered saline or other buffers, e.g., PBS or HEPES, to maintain pH at appropriate physiological levels. The expression constructs or vectors disclosed herein can be formulated into pharmaceuticalcompositions. These compositions may comprise, in addition to the vector, a pharmaceutically and / or physiologically acceptable excipient, carrier, buffer, stabilizer, antioxidants, preservative, or other additives well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient. The precise nature of the carrier or other material may be determined by the skilled person according to the route of administration. The pharmaceutical composition is typically in liquid form. Liquid pharmaceutical compositions generally include a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil. Additional carriers are provided in International Patent Publication No. W02000015822. Physiological saline solution, magnesium chloride, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol may be included. In some cases, a surfactant, such as pluronic acid (PF68) 0.001% may be used. In some cases, Ringer's Injection, Lactated Ringer's Injection, or Hartmann's solution is used. Preservatives, stabilizers, buffers, antioxidants and / or other additives may be included, as required.

[0184] Pharmaceutical compositions comprising an expression construct or vector disclosed herein may formulated with one or more pharmaceutically-acceptable excipients, which can be a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, carrier, manufacturing aid (e.g., lubricant, talc magnesium, calcium or zinc stearate, or steric acid), solvent or encapsulating material, involved in carrying or transporting the therapeutic compound for administration to the subject, bulking agent, salt, surfactant and / or a preservative. Some examples of materials which can serve as pharmaceutically-acceptable excipients include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; gelatin; talc; waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as ethylene glycol and propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents; water; isotonic saline; pH buffered solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.

[0185] A bulking agent is a compound which adds mass to a pharmaceutical formulation and contributes to the physical structure of the formulation in lyophilized form. Suitable bulking agents may include mannitol, glycine, polyethylene glycol and sorbitol.

[0186] The use of a surfactant can reduce aggregation and / or reduce the formation of particulates in the formulation. Suitable surfactants may include polysorbates e.g.,polysorbates 20 or 80); pol oxamers (e.g., pol oxamer 188); Triton; sodium dodecyl sulfate (SDS); sodium laurel sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl-or stearyl-sarcosine; linoleyl-, myristyl-, or cetylbetaine; lauroamidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-betaine (e.g., lauroamidopropyl); myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-dimethylamine; sodium methyl cocoyl-, or disodium methyl oleyl-taurate; and polyethyl glycol, polypropyl glycol, and copolymers of ethylene and propylene glycol (e.g., Pluronics, PF68, etc.).

[0187] Preservatives may be used in formulations and include octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride (a mixture of alkylbenzyl-dimethylammonium chlorides in which the alkyl groups are long-chain compounds), and benzethonium chloride. Other types of preservatives include aromatic alcohols such as phenol, butyl and benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol. Other suitable excipients can be found in standard pharmaceutical texts, e.g., in “Remington's Pharmaceutical Sciences,” The Science and Practice of Pharmacy, 19th Ed. Mack Publishing Company, Easton, Pa., (1995).

[0188] If a vector is to be stored long-term, it may be frozen in the presence of a cryoprotectant such as glycerol.

[0189] Methods of Treatment

[0190] Provided are methods of treating ocular disorders (including rod disorders) using the rod-specific promoters, expression constructs, vectors, or pharmaceutical compositions described herein. In one embodiment, the rod-specific promoters, expression constructs, vectors, or pharmaceutical compositions disclosed herein are used in a gene therapy to treat a subject who suffers from a rod disorder.

[0191] Examples of disorders that can be treated using the compositions and methods disclosed herein include retinal dystrophies. Conditions such as retinal dystrophies cause blindness due to destruction of the photoreceptors in the outer retina (i.e., the rods and cones). These conditions may be a result of direct damage to the photoreceptors, or photoreceptors being indirectly destroyed as a result of pathology in the retinal pigment epithelium and / or choroid. Severe visual impairment is common in advanced stages of the degeneration. Retinal dystrophies can be divided into rod-cone dystrophies (also called retinitis pigmentosa), conerod dystrophies and macular dystrophies. In rod-cone dystrophies the rod photoreceptorsdegenerate resulting in a loss of peripheral vision and night vision, and frequently this is followed by cone destruction leading to a loss of central and color vision. Conversely, in the cone-rod dystrophies there is initially a loss of cone photoreceptors leading to a loss of detailed and color vision and this is then followed by rod degeneration resulting in a loss of peripheral vision and night blindness. Both forms can result in blindness with extensive or complete loss of visual field. Another type of retinal dystrophy called macular dystrophy results in a loss of central vision, but peripheral vision is preserved. In some embodiments, the macular dystrophy is Stargardt disease, Vitelliform macular dystrophy, or North Carolina macular dystrophy.

[0192] In some embodiments, provided are methods of treating macular degeneration, including, but not limited to, age-related macular degeneration (AMD). AMD includes dry AMD, geographic atrophy (GA), and wet AMD.

[0193] In some embodiments, provided are methods of treating rod dystrophies such as congenital stational night blindness or retinitis pigmentosa.

[0194] Also provided are methods of prolonging the survival of cones that otherwise degenerate due to a bystander effect in a subject in need thereof using the rod-specific promoters, expression constructs, vectors, or pharmaceutical compositions disclosed herein. The bystander effect refers to a phenomenon where disease spreads from dying rods to healthy cones.

[0195] The terms “treat,” “treated,” “treating,” or “treatment” as used herein refer to therapeutic treatment, wherein the object is to slow down (lessen) an undesired physiological condition, disorder or disease, or to obtain beneficial or desired clinical results. For the purposes of this disclosure, beneficial or desired clinical results include, but are not limited to alleviation of symptoms; diminishment of the extent of the condition, disorder or disease; stabilization (z.e., not worsening) of the state of the condition, disorder or disease; delay in onset or slowing of the progression of the condition, disorder or disease; amelioration of the condition, disorder or disease state; and remission (whether partial or total), or enhancement or improvement of the condition, disorder or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Desirable treatment outcomes include, but are not limited to, the reduction or elimination of symptoms such as loss of visual acuity, abnormal color vision, photophobia, visual field loss, macular atrophy, rod degeneration, loss of cones, and / or loss of peripheral visual field.

[0196] Provided herein are method of treating disease (or preventing further progression of disease) in a subject in which cone function is wholly or partially reduced, e.g., at least 10%, at least 25%, at least 30%, at least 40%, at least 50%, at least 75%, at least 80%, at least 90%or more reduced as compared to a control. The control may be, for example, a healthy patient or group of healthy patients. The control may be the same patient at an earlier point in time. Healthy rods are rods that are capable of performing normal or partial, e.g., at least 10%, at least 25%, at least 30%, at least 50%, at least 75% or at least 90% of normal rod function in terms of perception of light at scotopic levels.

[0197] The terms “prevent”, “prevention”, and the like refer to acting prior to overt disease or disorder onset, to prevent the disease or disorder from developing or to minimize the extent of the disease or disorder or slow its course of development.

[0198] Also provided herein are methods of making rods more cone-like as disclosed in PCT Publication No. WO2016 / 135457, which is incorporated herein in its entirety, using a promoter, expression construct, vector, or pharmaceutical composition disclosed herein. In many mammalian species including mice and humans, the number of rod photoreceptors that mediate vision under dim light outnumbers greatly that of cone photoreceptors. However, in an industrialized world where illumination allows cones to operate throughout the day, rod-mediated vision can be less important. Many patients with absent rod function from birth are identified only incidentally and, in fact, cannot recognize their abnormal vision. On the contrary, when cone dysfunction is present, patients are always symptomatic and often suffer visual handicap dependent on the degree of their cone dysfunction. In some conditions, however, only (or mostly) the cones are lost or dysfunctional and rods remain relatively preserved. For example, achromatopsia is a severe hereditary retinal dystrophy with a complete absence of cone function from birth but, presumably, with a normal rod function. Mutations in multiple genes including CNGA3 and PDE6C have been associated with the disease. Each of the disease-causing genes encodes a component of the cone phototransduction cascade that translates light into an electric signal by causing hyperpolarization of the photoreceptor cell. In AMD, visual impairment is caused primarily by degeneration of the cone-rich fovea in the central macula. Thus, patients lose central vision and acuity, but often have relatively well-preserved peripheral macula and thus have some useful residual vision that is limited by the paucity of cones outside the fovea. Accordingly, provided herein are methods of making rods more cone-like by administering to a subject a vector comprising a nucleic acid comprising a rods-specific promoter disclosed herein and an operatively linked transgene, wherein the transgene encodes a gene product that is light-sensitive and / or that modulates endogenous light-sensitive signaling in a photoreceptor cell, whereby expression of the transgene in a rod photoreceptor cell increases the intensity of the light to which the rod photoreceptor responds and / or increases the speed at which the rod photoreceptor responds to light. In embodiments,the gene product is ArchT, R9AP, Jaws, iClC2, Rhodopsin, GNAT1, CNGB1, APOE, HTRA1, or C3.

[0199] Provided is a method of increasing the intensity of the light to which a rod photoreceptor responds, the method comprising contacting the rod photoreceptor with an expression construct, a vector, or a pharmaceutical composition disclosed herein.

[0200] Provided is a method of increasing the speed at which a rod photoreceptor responds to light, the method comprising contacting the rod photoreceptor with an expression construct, a vector, or a pharmaceutical composition disclosed herein.

[0201] Provided herein is a method of treating a subject with a rod disorder, the method comprising administering to the subject an expression construct, a vector, or a pharmaceutical composition disclosed herein, wherein the administering increases the sensitivity of the subject’s rod cells.

[0202] Provided herein is a method of prolonging survival of cones in a subject in need thereof, the method comprising administering to the subject an expression construct, a vector, or a pharmaceutical composition disclosed herein, wherein the administering increases the sensitivity of the subject’s rod cells.

[0203] Provided herein are methods of treatment, wherein a therapeutically effective amount of an expression construct, a vector, or a pharmaceutical composition disclosed herein is administered to a subject. An “effective amount” or “therapeutically effective amount” refers to an amount of the compound or agent that is capable of producing a medically desirable result in a treated subject. The treatment method can be performed alone or in conjunction with other drugs or therapy. A therapeutically effective amount can be administered in one or more administrations, applications or dosages and is not intended to be limited to a particular formulation or administration route.

[0204] As used herein, the terms “subject” and “patient” are used interchangeably irrespective of whether the subject has undergone treatment in the past or is currently undergoing any form of treatment. As used herein, the terms “subject” and “subjects” may refer to any vertebrate, including, but not limited to, a mammal (e.g., cow, pig, camel, llama, horse, goat, rabbit, sheep, hamsters, guinea pig, cat, dog, rat, and mouse, a non-human primate (for example, a monkey, such as a cynomolgus monkey, chimpanzee, etc.) or a human). The subject may be a human or a non-human. In some embodiments, the subject is a human.

[0205] A person skilled in the art may choose a suitable route of administration. In embodiments, administration of the expression construct, vector, or pharmaceutical composition disclosed herein is by direct retinal, subretinal or intravitreal injection.Administration includes direct delivery to photoreceptor cells. A vector disclosed herein may transduce the target cells (e.g., rod photoreceptors) without entering significant numbers of other cell populations (e.g., cone photoreceptors).

[0206] The expression construct, vector, or pharmaceutical composition disclosed herein may be administered in order to prevent or delay the onset of one or more symptoms of an ocular disorder. The subject may be asymptomatic or may have a predisposition to the disease. The method or use may comprise a step of identifying a subject that has, or is at risk of developing, an ocular disorder.

[0207] The dose of a vector of the invention may be determined according to various parameters, especially according to the age, weight and condition of the patient to be treated; the route of administration; and the required regimen. A typical single dose is about 108, about 109, about 1010, about 1011, about 1012, about 1012, about 1013, or about 1014, genome particles, depending on the amount of retinal tissue that requires transduction. A genome particle is defined herein as an AAV capsid that contains a viral genome that can be quantified with a sequence specific method (such as real-time PCR). That dose may be provided as a single dose, but may be repeated for the fellow eye or in cases where vector may not have targeted the correct region of retina for whatever reason (such as surgical complication). The treatment is preferably a single permanent treatment for each eye, but repeat injections, for example in future years and / or with different AAV serotypes may be considered.

[0208] In some embodiments, AAV particles disclosed herein are provided in a volume of between about 5 pL to about 5 mL. In some embodiments, AAV particles are provided in a volume of between about 20 pL to about 3 mL. In some embodiments, AAV particles are provided in a volume of about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 200, about 300, about 400, about 500, about 600, about 700, about 800, about 900, about 1000 pL.

[0209] In embodiments, the expression construct, vector, or pharmaceutical composition disclosed herein is administered with a device described in any one of PCT Publications Nos. WO2018 / 112305, W02021 / 055906, and WO2022 / 036256, which are incorporated by reference herein in their entireties.

[0210] Kits

[0211] The expression constructs, vectors and / or pharmaceutical compositions disclosed herein can be packaged into a kit. Provided are kits for the manufacturing, preparation, and development of the compositions described above, comprising at least one or more containers,each with a different reagent for the manufacturing of the compositions disclosed herein. Kits may include a set of instructions in the use of the reagents, important information on how to perform the procedures for the manufacturing.

[0212] In certain embodiments, kits are provided for the preparation of a pharmaceutical composition comprising a therapeutically effective amount of a composition disclosed herein and a pharmaceutically acceptable carrier. The kit may comprise at least one, or more containers, each with a different reagent. Kits may include instruction for the preparation, for the therapeutic regimen to be used, and periods of administration.

[0213] The composition or the pharmaceutical composition described herein can be provided in a kit. In one embodiment, the kit includes (a) a container that contains the composition and optionally (b) informational material. The informational material can be descriptive, instructional, marketing or other material that relates to the methods described herein and / or the use of the agents for therapeutic benefit. For example, kits may include instruction for the preparation and / or administration of the compositions described herein. In an embodiment, the kit also includes an additional therapeutic agent. The kit may comprise one or more containers, each with a different reagent.

[0214] The containers can include a unit dosage of the pharmaceutical composition. In addition to the composition, the kit can include other ingredients, such as a solvent or buffer, an adjuvant, a stabilizer, or a preservative.

[0215] The kit optionally includes a device suitable for administration of the composition, e.g., a syringe or other suitable delivery device. The device can be provided pre-loaded with one or both of the agents or can be empty, but suitable for loading.

[0216] It is to be understood that this invention is not limited to the particular molecules, compositions, methodologies, or protocols described, as these may vary. Any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present invention. It is further to be understood that the disclosure of the invention in this specification includes all possible combinations of such particular features. For example, where a particular feature is disclosed in the context of a particular aspect or embodiment of the invention, or a particular claim, that feature can also be used, to the extent possible, in combination with and / or in the context of other particular aspects and embodiments of the invention, and in the invention generally.

[0217] Where reference is made herein to a method comprising two or more defined steps, the defined steps can be carried out in any order or simultaneously (except where the contextexcludes that possibility), and the method can include one or more other steps which are carried out before any of the defined steps, between two of the defined steps, or after all the defined steps (except where the context excludes those possibilities).

[0218] All referenced patents and applications, scientific articles, book chapters, and other references are incorporated herein by reference in their entireties. Furthermore, where a definition or use of a term in a reference, which is incorporated by reference herein is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply.

[0219] To facilitate a better understanding of the present invention, the following Examples of specific embodiments are given. The following examples should not be read to limit or define the entire scope of the invention.EXAMPLES

[0220] Example 1. Materials and Methods

[0221] This Example details the materials and methods used in Examples 1-5.

[0222] Vector design and production

[0223] Rod-specific promoters were rationally designed by intersecting multiple genomic and epigenomic datasets. Several types of data were used, including DNA accessibility assays, transcription factor binding affinity, histone markers, validated transcription start sites and regions with cis-regulatory elements predicted to contact the promoter of interest (FIG. 1). To pinpoint the genomic regions with active promoter activity in rod cells, the analysis was restricted to a 10 kb upstream- 1 kb downstream region of the transcription start sites (TSS) of interest. The selection of suitable TSS was performed by cross-referencing current literature and single cell sequencing data from the human retina. Regions harboring an increased number of regulatory elements were isolated and combined with their target core promoter.

[0224] Additionally, following an enrichment analysis for transcription factors regulating a set of rod-specific genes (TRANSFAC / MAST), a preferred set of transcription factor binding sites (TFBS) was developed in silico and predicted to have a significant impact in regulating a rod-specific promoter when placed withing the same genomic context. Designed promoter sequences were then cloned into an AAV backbone carrying eGFP and finally packaged into AAV vectors with AAV5 or AAV7m8 capsids, alongside vectors carrying the commonly used RK promoter driving eGFP expression (control).

[0225] All vectors were titred using a qPCR-based titration assay targeting the BGH polyA sequence following a set PCR protocol as known in the art. In particular, a TaqMan probe was used in this qPCR method. Table 1 provides a summary of all vectors used in this study.

[0226] Vector nomenclature (see Table 1): AAV genome / AAV capsid - promoter -transgene. For example, an AAV2 / 7m8-RK-eGFP vector comprises ITRs derived from AAV2 (z.e., an AAV2 genome), capsid proteins derived from AAV 7m8, and comprises an eGPF gene under the control of a RK promoter.

[0227] Expression construct nomenclature (see Table 1): promoter - transgene - post-transcriptional regulatory element - polyadenylation (pA) sequence.Table 1. Promoter and vector constructs used to assess rod-specific promoter activity. All vectors used an AAV2 genome. See also FIG. 7 and Tables 3 and 6. _ _ Promoter Composition of expression AAV Vector name Titre name construct capsid (VG / ml) RK RK-eGFP-WPRE-BGHpA 7m8 AAV2 / 7m8-RK- 6.65E+13 eGFPAAV5 AAV2 / 5-RK- 1.19E+13 eGFP1 Prom 1-eGFP-WPRE- 7m8 AAV2 / 7m8- 1.42E+13 BGHpA Prom 1-eGFPAAV5 AAV2 / 5-Prom 7.36E+131-eGFP2 Prom 2-eGFP-WPRE- 7m8 AAV2 / 7m8- 8.92E+13 BGHpA Prom 2-eGFPAAV5 AAV2 / 5- Prom 1.4E+142-eGFP3 Prom 3-eGFP-WPRE- 7m8 AAV2 / 7m8- 1.29E+13 BGHpA Prom 3-eGFPAAV5 AAV2 / 5-Prom 1.46E+143-eGFP4 Prom 4-eGFP-WPRE- 7m8 AAV2 / 7m8- 7.32E+13 BGHpA Prom 4-eGFPAAV5 AAV2 / 5- Prom 1.35E+144-eGFP5 (“V5”) Prom 5-eGFP-WPRE- 7m8 AAV2 / 7m8- 1.25E+13 BGHpA Prom 5- eGFPAAV5 AAV2 / 5- Prom 4.29E+145-eGFP6 Prom 6-eGFP-WPRE- 7m8 AAV2 / 7m8- 7.23E+13 BGHpA Prom 6-eGFPAAV5 AAV2 / 5- Prom 1.08E+146-eGFP7 Prom 7-eGFP-WPRE- 7m8 AAV2 / 7m8- 1.31E+13BGHpA Prom 7-eGFPAAV5 AAV2 / 5-Prom 5.83E+147-eGFP8 Prom 8-eGFP-WPRE- 7m8 AAV2 / 7m8- 4.96E+13 BGHpA Prom 8 -eGFPAAV5 AAV2 / 5- Prom 2.59E+148-eGFP9 Prom 9-eGFP-WPRE- 7m8 AAV2 / 7m8- 3.54E+13 BGHpA Prom9-eGFPAAV5 AAV2 / 5-Prom9- 8.29E+13 eGFP10 Prom 10-eGFP-WPRE6- 7m8 AAV2 / 7m8- 1.04E+13 bGHpA Prom 10-eGFPAAV5 AAV2 / 5-Prom 4.04E+1410-eGFP

[0228] Generation of wild-type retinal organoids

[0229] Wild-type (WT) retinal organoids (ROs) were generated from human induced pluripotent stem cells (IPSCs) and human embryonic stem cells (HESCs) using a differentiation protocol as known in the art.

[0230] Transduction of wild-type retinal organoids

[0231] At differentiation day 130 to 170, WT ROs with laminated morphology were selected for transduction and transferred to an individual well of a 24 well plate. Organoids were dosed with 1E11 viral genomes (VGs) per organoid (AAV2 / 7m8) in a volume of 100 pl. The medium was topped up to 200 pl by the end of the day. The next day, media was topped up by an additional 800 pl. A complete media change was carried out 72 hours later. Organoids were harvested for flow cytometric or immunofluorescent staining 21 days post transduction. Three separate transduction experiments were carried out with a final n = 5 retinal organoids for flow cytometry (FC) analysis and n = 2 for immunohistochemical (IHC) analysis.

[0232] Flow cytometric analysis of eGFP intensity in retinal organoids

[0233] Three weeks post transduction, organoids were dissociated into single cells and stained with photoreceptor specific marker CD73 conjugated to APC (Allophycocyanin) (Table 2) for fluorescence-activated cell sorting (FACS) analysis as known in the art.Table 2. Materials used in the study.Antibody Host Company (Catalogue Dilution Application target species #)L / M Opsin Rabbit Millipore AB5405 1:500 IHC (humanorganoids)eGFP Chicken AB16901 1:500 IHC (human organoids) CD73-APC Mouse Milteny 130-123-802 1:50 FC (human organoids) Biotinylated n / a Vector Laboratories 1:500 IHC (mouse retina) Peanut (B-1075-5)Agglutinin(PNA)Streptavidin n / a Thermo Fisher 1:500 IHC (mouse retina) AF546 (SI 1225)DAPI n / a Invitrogen (D21490) 1:2000 IHC (mouse retina,human organoids)Table 3. Illustrative promoter sequences. See also Table 6 for additional sequence information.SEQ Promoter SequenceID NO:17 Component aaaccagaaagtctctagctgtccagaggacatagcacagaggcccatggtccctatttcaaacc B caggccaccagactgagctgggaccttgggacagacaagtcatgcagaagttaggggaccttct cctcccttttcctggatcctgagtacctctcctccctgacctcaggcttcctcctagtgtcaccttggc ccctcttagaagccaattaggccctcagtttctgcagcggggattaatatgattatgaacaccccca atctcccagatgctgattcagccaggagcttaggagggggaggtcactttataagggtctgggg gggtcagaacccagagtcatc18 Component acagagtggagaaccgaagcacactctcgggagggcggggtagctggctgcacccaggctgt Al gtagcctcagtccagatgtaagggtgggtggaaaagagccttgcccaatgagggagaacagtg aaaccaaggccatagggtctaaagattcacgaaccaggctctcatggagaaagcaggtgaggtt tactgtatagatgggtgtgcccctaccccacactgaggcttcctcgtctgagcaaactgaggccca gagaggggaaggaagcaggactaccatggtgactcaaagaccagctagaatccagcctcctct cctcgaggcttccactgccccacgccaggcctgtgtgactcagtctagggcctttccattacccca gctaaacctttctttagtcatttataccatggtgtgaatggctggctggtctttcctgagagctatcttt gatgaggggagggaggcatagccaggtttgggaagctgataccccaggaagcccagttgact gtgtgggttatagcccaggctgtcactgatttgtaacgggacct19 Component ctataatcctggcactttgggaggccgaggagggtggatcgcttgagccaggagttcaggacca A2 gcctgggcaacatagcgagactccacccctacaaaaaatacaaaaactagtggtgtgcacttgtg gtcccagctactcaggaggctgaggtgagaggatcgcttgagcccaggaggcagaggctaca gtgagctatgattgtggcactgcactccagcctgggcgacagagaccttgtctcaaaacttttttttt cttcgtcaagctttacagaataaagagcactgtcacctcagtgatggctgttagttccccatcacca gggctccatgaggttgcaattgtgaaactcacaaaggaggaacctgagacagagaggggaagt actgagatcatctaggtccattcccccactcactcgttcattcaacaaatattcaggagcaccttcta ggtgccaggccctggagacacatcagtgaacaaaacagacatcatcccacctctttccactacag gccaagcaccatgctggtctctgggaaccctgttgtgagcaagacagacccaggcttacccttgt ggactcatgttacaggcagggagacgggcacaaaacacaaataaaaagcttccatgctgtcaga agcactatgcaaaaagcaagatgctgaggtactgctaagctgtgtgggatgggggctcagcccg gccaggga20 Component caatgatggagaccggggcggggtccagaggtaacttgtgccccgccccgatagcctagggc C ccagcggggattaagttttttttttttttcttttgaggaatcttactccaatgatggcgcctgctgaattcgaggaatcttactagaccggggcggggccacataaggttgcaaatcccacagagagcctgcgcSEQ Promoter SequenceID NO:ctggggttgttttctgtaggcgcctgtgcccccgcccctgccaccaatgatggcgcctgctgaatt c21 Component gaagggttgggggggctgagctggattcacctgtccttgtctctgattggctcttggacaccccta D gcccccaaatcccactaagcagccccaccagggattgcacaggtccgtagagagccagttgatt gcaggtcctcctggggccagaagggtgcctgggaggccaggttctggggatcccctccatcca gaagaaccacctgctcactctgtcccttcgcctgctgctgggac27 Component ccaatgatggagaccggggcggggtccagaggtaacttgtgccccgccccgatagcctaggg E cccagcggggattaagttttttttttttttcttttgaggaatcttactccaatgatggcgcctgctcaatt cgaggaatcttactagaccggggcggggccacataaggttgcaaatcccacagagagcctgcg cctggggttgttttctgtaggcgcctgtgcccccgcccctgccaccaatgatggcgcctgctcaat tcctatgtgtctggcaccagaaacggaagctgcaggttgcagcccctgccctcatggagctcctc ctgtcagaggagtgtggggactggatgactccagaggtaacttgtgggggaacgaacaggtaa ggggctgtgtgacgagatgagagactgggagaat28 Component aaaccagaaagtctctagctgtccagaggacatagcacagaggcccatggtccctatttcaaacc F caggccaccagactgagctgggaccttgggacagacaagtcatgcagaagttaggggaccttct cctcccttttcctggatactgagtacctctcctccctgacctcaggcttcctcctagtgtcaccttggc ccctcttagaagccaattaggccctcagtttctgcagcggggattaatatgattatgaacaccccca atctcccagatgctgattcagccaggagcttaggagggg29 Component gaggtcactttataagggtctgggggggtcagaacccctagttcatccagctggagcgagccga G acggtcgttgagccaagaggtaagggtttaagggatggttggttggtggggtattaatgtttaatta cctggagcacctgcctgaaatcactttttttcaggttggccgcggatccgccacc30 Component cggcgcctatataaggtgtgggcgggtgtggcacagctagttccgtcgcagccggcgagccga H acggtcgttgagccaagaggtaagggtttaagggatggttggttggtggggtattaatgtttaatta cctggagcacctgcctgaaatcactttttttcaggttggccgcggatccgccacc31 Component tgtagttaatgattaacccgccatgctacttatctacgtacatttatattggctcatgtccaacattacc I gccatgttgacattgattattgactagaattcgctagcaagatccaagctcagatctcgatcgagttg ggccccagaagcctggtggttgtttgtccttctcaggggaaaagtgaggcggccccttggagga aggggccgggcagaatgatctaatcggattccaagcagctcaggggattgtctttttctagcacct tcttgccactcctaagcgtcctccgtgaccccggctgggatttagcctggtgctgtgtcagccccg gtctcccaggggcttcccagtggtccccaggaaccctcgacagggcccggtctctctcgtccag caagggcagggacgggccacaggccaagggccctcgatcgaggaactgaaaaaccagaaa gttaactggtaagtttagtagaccggggcggggtccagaggtaacttgtgccccgccccgatag cctagggcccagcggggattaagttttttttttttttcttttgaggaatcttactccaatgatggcgcct gctcaattcgaggaatcttactagaccggggcggggccacataaggttgcaaatcccacagaga gcctgcgcctggggttgttttctgtaggcgcctgtgcccccgcccctgccaccaatgatggcgcc tgctcaattcctatgtgtctggcaccagaaacggaagctgcaggttgcagcccctgccctcatgg agctcctcctgtcagaggagtgtggggactggatgactccagaggtaacttgtgggggaacgaa cgcgtaaggggctgtgtgacgagatgagagactgggagaat32 Component gaggtcactttataagggtctgggggggtcagaacccagagtcatccagctggagccctgagtg J gctgagctcaggccttcgcagcattcttgggtgggagcagccacgggtcagccacaagggcca cagccctttttgtcttttatttcaggtcccggatccggtggtggtgcaaatcaaagaactgctcctca gtggattttgcctttacttctaggcctgtacggaagtgttacttctgctctaaaagctgcggaattgta cccgcggccgc33 Component ctatgtgtctggcaccagaaacggaagctgcaggttgcagcccctgccctcatggagctcctcct K gtcagaggagtgtggggactggatgactccagaggtaacttgtgggggaacgaacaggtaaggggctgtgtgacgagatgagagactgggagaatSEQ Promoter SequenceID NO:34 Component cagctggagccctgagtggctgagctcaggccttcgcagcattcttgggtgggagcagccacg L ggtcagccacaagggccacagcc35 Component tccctgcaggtcataaaatcccagtccagagtcaccagcccttcttaaccacttcctactgtgtgac M cctttcagcctttacttcctcatcagtaaaatgaggctgatgatatgggcatccatactccagggcc agtgtgagcttacaacaagataaggagtggtgctgagcctggtgccgggcaggcagcaggcat gtttctcccaattatgccctctcactgccagccccacctccattgtcctcacccccagggctcaagg ttctgccttcccctttctcagccctgaccctactgaacatgtctccccactcccaggcagtgccagg gcctctcctggagggttgcggggacagaaggacagccggagtgcagagtcagcggttgaggg attggggctatgccagctaatcc36 Component gctgcccaggccacagccgcctttgggttccatcttgctaataaacactggctctgggactagact N ttggcttctctccttggtcaggggatttagagccacacgtggtgggcctggggagccaggtggg gggctgggaagggagtagattggctccatttaaagaaacccaacatgtagaaaccacatttagttt gacaaacctgggctgctgcaccaccccaaacacttgggtagtgcgaagggctgtagagacccc tcagggcccactcactcctcagggtcagggagggtgggtggagcctgaaggctgaaagtggc agaaatgcaacggcctcactcctggacagatccccatcatccaacagggtgggtgtagaagctg ccagcctgtggtaggagccagcattcacttcccacgcatactatttagagacagccgttagtgctc agtgtctggtgccccggcacccccttgtggtctcttgagtctgtggcagagttgagtgctgaaagg cagccaggttccagggaagagtccagccaggaggctcagggtgcctggaggagcctttgaga gcccaagatttgggttttcctatggaagttgagggtgggaagtctctagaacctgggagctcaacc tgaacaatttctgcccggcagtgagccctgcacctgaacagagcaccagaccccctaagctgag gcaggcaggccctggagataatcttaggcaggcctcagagcacccacagcagacagagcagg agcaggataggtgagccccattcctccataccccagggaacactacacctgctctgggatgcaat ttctccacctggcaacaggcctgcaaaacattctctggaagagcctggccacttctgcagattaga gactttgttgcacccaaattcaagtgtacatataggcacaggcgcctgcaagcatgcatgcacac acacagtcatacacaggcagagatggacatgcacaggcacacatatgctcaggtacaaacagg catgtttacacccctgcgtattccctgcaggtcataaaatcccagtccagagtcaccagcccttctt aaccacttcctactgtgtgaccctttcagcctttacttcctcatcagtaaaatgaggctgatgatatgg gcatccatactccagggccagtgtgagcttacaacaagataaggagtggtgctgagcctggtgc cgggcaggcagcaggcatgtttctcccaattatgccctctcactgccagccccacctccattgtcc tcacccccagggctcaaggttctgccttcccctttctcagccctgaccctactgaacatgtctcccc actcccaggcagtgccagggcctctcctggagggttgcggggacagaaggacagccggagtg cagagtcagcggttgagggattggggctatgccagctaatcc37 Component agtcagcggttgagggattggggctatgccagcccgatta01 1 ccatcctcaaggcccaggcctgggacacagcctttctggcagagcccaaggcagggctgctga gaccctgcccagagaagactctaagccagaaagaacctgcctgcgtcctggcctgggggctgg agagttggggtacacctcttgccaccgctgagtgggagctgacaagcccctgctaccatgagga gtctccaggcatcacatgggaactccaagctgttattctcccatcagccactccgtgtcttggccaa ggtgtatgggaagccaggcccctggtaagccgggcaggtcccctggcaacccatggtgctcca aacctgggccatgcaggagcccttccctggggcccacatccttgctggaccctgggagttggga ggtgctgctggtttcctgggcatcaacagcctcttttctccccctcccccattcagcacctgctaatc ctttgcacacggcccagctactgccaattaatcttccttgtgtctgaggggccagctacgagtggc agcaagaaggcaattcctggctggcggttggcatctaagcag5 2 tctgtgcagacatctcttctttgctcttactagaccaaggtgaaagaaaactctcaccttctcccatct ggccccacagcatctggaacacactgatcctcataatccttgttcttgagaaatattaatgacttaat ctcccaagcttgctccctctcctgtgcaggccatctcagtatgttttgcagacaagacccagagaagtccaSEQ Promoter SequenceID NO:10 3 ccaatgatggagaccggggcggggtccagaggtaacttgtgccccgccccgatagcctaggg cccagcggggattaagttttttttttttttcttttgaggaatcttactccaatgatggcgcctgctgaatt cgaggaatcttactagaccggggcggggccacataaggttgcaaatcccacagagagcctgcg cctggggttgttttctgtaggcgcctgtgcccccgcccctgccaccaatgatggcgcctgctgaat tcctatgtgtctggcaccagaaacggaagctgcaggttgcagcccctgccctcatggagctcctc ctgtcagaggagtgtggggactggatgactccagaggtaacttgtgggggaacgaacaggtaa ggggctgtgtgacgagatgagagactgggagaataaaccagaaagtctctagctgtccagagg acatagcacagaggcccatggtccctatttcaaacccaggccaccagactgagctgggaccttg ggacagacaagtcatgcagaagttaggggaccttctcctcccttttcctggatcctgagtacctctc ctccctgacctcaggcttcctcctagtgtcaccttggcccctcttagaagccaattaggccctcagtt tctgcagcggggattaatatgattatgaacacccccaatctcccagatgctgattcagccaggagc ttaggagggggaggtcactttataagggtctgggggggtcagaacccagagtcatccagctgga gccctgagtggctgagctcaggccttcgcagcattcttgggtgggagcagccacgggtcagcc acaagggccacagcc4 4 tccctgcaggtcataaaatcccagtccagagtcaccagcccttcttaaccacttcctactgtgtgac cctttcagcctttacttcctcatcagtaaaatgaggctgatgatatgggcatccatactccagggcc agtgtgagcttacaacaagataaggagtggtgctgagcctggtgccgggcaggcagcaggcat gtttctcccaattatgccctctcactgccagccccacctccattgtcctcacccccagggctcaagg ttctgccttcccctttctcagccctgaccctactgaacatgtctccccactcccaggcagtgccagg gcctctcctggagggttgcggggacagaaggacagccggagtgcagagtcagcggttgaggg attggggctatgccagctaatccgaagggttgggggggctgagctggattcacctgtccttgtctc tgattggctcttggacacccctagcccccaaatcccactaagcagccccaccagggattgcaca ggtccgtagagagccagttgattgcaggtcctcctggggccagaagggtgcctgggaggccag gttctggggatcccctccatccagaagaaccacctgctcactctgtcccttcgcctgctgctggga ccgcggccgc3 5 gctgcccaggccacagccgcctttgggttccatcttgctaataaacactggctctgggactagact ttggcttctctccttggtcaggggatttagagccacacgtggtgggcctggggagccaggtggg gggctgggaagggagtagattggctccatttaaagaaacccaacatgtagaaaccacatttagttt gacaaacctgggctgctgcaccaccccaaacacttgggtagtgcgaagggctgtagagacccc tcagggcccactcactcctcagggtcagggagggtgggtggagcctgaaggctgaaagtggc agaaatgcaacggcctcactcctggacagatccccatcatccaacagggtgggtgtagaagctg ccagcctgtggtaggagccagcattcacttcccacgcatactatttagagacagccgttagtgctc agtgtctggtgccccggcacccccttgtggtctcttgagtctgtggcagagttgagtgctgaaagg cagccaggttccagggaagagtccagccaggaggctcagggtgcctggaggagcctttgaga gcccaagatttgggttttcctatggaagttgagggtgggaagtctctagaacctgggagctcaacc tgaacaatttctgcccggcagtgagccctgcacctgaacagagcaccagaccccctaagctgag gcaggcaggccctggagataatcttaggcaggcctcagagcacccacagcagacagagcagg agcaggataggtgagccccattcctccataccccagggaacactacacctgctctgggatgcaat ttctccacctggcaacaggcctgcaaaacattctctggaagagcctggccacttctgcagattaga gactttgttgcacccaaattcaagtgtacatataggcacaggcgcctgcaagcatgcatgcacac acacagtcatacacaggcagagatggacatgcacaggcacacatatgctcaggtacaaacagg catgtttacacccctgcgtattccctgcaggtcataaaatcccagtccagagtcaccagcccttctt aaccacttcctactgtgtgaccctttcagcctttacttcctcatcagtaaaatgaggctgatgatatgg gcatccatactccagggccagtgtgagcttacaacaagataaggagtggtgctgagcctggtgc cgggcaggcagcaggcatgtttctcccaattatgccctctcactgccagccccacctccattgtcc tcacccccagggctcaaggttctgccttcccctttctcagccctgaccctactgaacatgtctccccactcccaggcagtgccagggcctctcctggagggttgcggggacagaaggacagccggagtgSEQ Promoter SequenceID NO:cagagtcagcggttgagggattggggctatgccagctaatccgaagggttgggggggctgagc tggattcacctgtccttgtctctgattggctcttggacacccctagcccccaaatcccactaagcag ccccaccagggattgcacaggtccgtagagagccagttgattgcaggtcctcctggggccagaa gggtgcctgggaggccaggttctggggatcccctccatccagaagaaccacctgctcactctgt cccttcgcctgctgctgggac cgcggccgc8 6 acagagtggagaaccgaagcacactctcgggagggcggggtagctggctgcacccaggctgt gtagcctcagtccagatgtaagggtgggtggaaaagagccttgcccaatgagggagaacagtg aaaccaaggccatagggtctaaagattcacgaaccaggctctcatggagaaagcaggtgaggtt tactgtatagatgggtgtgcccctaccccacactgaggcttcctcgtctgagcaaactgaggccca gagaggggaaggaagcaggactaccatggtgactcaaagaccagctagaatccagcctcctct cctcgaggcttccactgccccacgccaggcctgtgtgactcagtctagggcctttccattacccca gctaaacctttctttagtcatttataccatggtgtgaatggctggctggtctttcctgagagctatcttt gatgaggggagggaggcatagccaggtttgggaagctgataccccaggaagcccagttgact gtgtgggttatagcccaggctgtcactgatttgtaacgggacctaaaccagaaagtctctagctgt ccagaggacatagcacagaggcccatggtccctatttcaaacccaggccaccagactgagctg ggaccttgggacagacaagtcatgcagaagttaggggaccttctcctcccttttcctggatcctga gtacctctcctccctgacctcaggcttcctcctagtgtcaccttggcccctcttagaagccaattagg ccctcagtttctgcagcggggattaatatgattatgaacacccccaatctcccagatgctgattcag ccaggagcttaggagggggaggtcactttataagggtctgggggggtcagaacccagagtcat c6 7 ctatgtgtctggcaccagaaacggaagctgcaggttgcagcccctgccctcatggagctcctcct gtcagaggagtgtggggactggatgactccagaggtaacttgtgggggaacgaacaggtaagg ggctgtgtgacgagatgagagactgggagaataaaccagaaagtctctagctgtccagaggac atagcacagaggcccatggtccctatttcaaacccaggccaccagactgagctgggaccttggg acagacaagtcatgcagaagttaggggaccttctcctcccttttcctggatcctgagtacctctcct ccctgacctcaggcttcctcctagtgtcaccttggcccctcttagaagccaattaggccctcagtttc tgcagcggggattaatatgattatgaacacccccaatctcccagatgctgattcagccaggagctt aggagggggaggtcactttataagggtctgggggggtcagaacccagagtcatccagctggag ccctgagtggctgagctcaggccttcgcagcattcttgggtgggagcagccacgggtcagccac aagggccacagcc7 8 agaagccaattaggccctcagtttctgcagcggggattaatatgattatgaacacccccaatctcc cagatgctgattcagccaggagcttaggagggg9 9 ctataatcctggcactttgggaggccgaggagggtggatcgcttgagccaggagttcaggacca gcctgggcaacatagcgagactccacccctacaaaaaatacaaaaactagtggtgtgcacttgtg gtcccagctactcaggaggctgaggtgagaggatcgcttgagcccaggaggcagaggctaca gtgagctatgattgtggcactgcactccagcctgggcgacagagaccttgtctcaaaacttttttttt cttcgtcaagctttacagaataaagagcactgtcacctcagtgatggctgttagttccccatcacca gggctccatgaggttgcaattgtgaaactcacaaaggaggaacctgagacagagaggggaagt actgagatcatctaggtccattcccccactcactcgttcattcaacaaatattcaggagcaccttcta ggtgccaggccctggagacacatcagtgaacaaaacagacatcatcccacctctttccactacag gccaagcaccatgctggtctctgggaaccctgttgtgagcaagacagacccaggcttacccttgt ggactcatgttacaggcagggagacgggcacaaaacacaaataaaaagcttccatgctgtcaga agcactatgcaaaaagcaagatgctgaggtactgctaagctgtgtgggatgggggctcagcccg gccagggaaaaccagaaagtctctagctgtccagaggacatagcacagaggcccatggtccct atttcaaacccaggccaccagactgagctgggaccttgggacagacaagtcatgcagaagttag gggaccttctcctcccttttcctggatcctgagtacctctcctccctgacctcaggcttcctcctagtgtcaccttggcccctcttagaagccaattaggccctcagtttctgcagcggggattaatatgattatgaSEQ Promoter SequenceID NO:acacccccaatctcccagatgctgattcagccaggagcttaggagggggaggtcactttataagg gtctgggggggtcagaacccagagtcatc2 10 agtcagcggttgagggattggggctatgccagcccgattagaagggttgggggggctgagctg gattcacctgtccttgtctctgattggctcttggacacccctagcccccaaatcccactaagcagcc ccaccagggattgcacaggtccgtagagagccagttgattgcaggtcctcctggggccagaag ggtgcctgggaggccaggttctggggatcccctccatccagaagaaccacctgctcactctgtcc cttcgcctgctgctgggac22 V12 (Enhl- acagagtggagaaccgaagcacactctcgggagggcggggtagctggctgcacccaggctgt TFBS- gtagcctcagtccagatgtaagggtgggtggaaaagagccttgcccaatgagggagaacagtg Rho(MD)- aaaccaaggccatagggtctaaagattcacgaaccaggctctcatggagaaagcaggtgaggtt MVMi) tactgtatagatgggtgtgcccctaccccacactgaggcttcctcgtctgagcaaactgaggccca gagaggggaaggaagcaggactaccatggtgactcaaagaccagctagaatccagcctcctct cctcgaggcttccactgccccacgccaggcctgtgtgactcagtctagggcctttccattacccca gctaaacctttctttagtcatttataccatggtgtgaatggctggctggtctttcctgagagctatcttt gatgaggggagggaggcatagccaggtttgggaagctgataccccaggaagcccagttgact gtgtgggttatagcccaggctgtcactgatttgtaacgggacctccaatgatggagaccggggcg gggtccagaggtaacttgtgccccgccccgatagcctagggcccagcggggattaagttttttttt tttttcttttgaggaatcttactccaatgatggcgcctgctcaattcgaggaatcttactagaccggg gcggggccacataaggttgcaaatcccacagagagcctgcgcctggggttgttttctgtaggcg cctgtgcccccgcccctgccaccaatgatggcgcctgctcaattcctatgtgtctggcaccagaa acggaagctgcaggttgcagcccctgccctcatggagctcctcctgtcagaggagtgtggggac tggatgactccagaggtaacttgtgggggaacgaacaggtaaggggctgtgtgacgagatgag agactgggagaataaaccagaaagtctctagctgtccagaggacatagcacagaggcccatgg tccctatttcaaacccaggccaccagactgagctgggaccttgggacagacaagtcatgcagaa gttaggggaccttctcctcccttttcctggatactgagtacctctcctccctgacctcaggcttcctcc tagtgtcaccttggcccctcttagaagccaattaggccctcagtttctgcagcggggattaatatga ttatgaacacccccaatctcccagatgctgattcagccaggagcttaggagggggaggtcacttt ataagggtctgggggggtcagaacccctagttcatccagctggagcgagccgaacggtcgttg agccaagaggtaagggtttaagggatggttggttggtggggtattaatgtttaattacctggagca cctgcctgaaatcactttttttcaggttggccgcggatccgccacc23 V13 ctataatcctggcactttgggaggccgaggagggtggatcgcttgagccaggagttcaggacca (Enh2- gcctgggcaacatagcgagactccacccctacaaaaaatacaaaaactagtggtgtgcacttgtg TFBS- gtcccagctactcaggaggctgaggtgagaggatcgcttgagcccaggaggcagaggctaca Rho(MD)- gtgagctatgattgtggcactgcactccagcctgggcgacagagaccttgtctcaaaacttttttttt MVMi) cttcgtcaagctttacagaataaagagcactgtcacctcagtgatggctgttagttccccatcacca gggctccatgaggttgcaattgtgaaactcacaaaggaggaacctgagacagagaggggaagt actgagatcatctaggtccattcccccactcactcgttcattcaacaaatattcaggagcaccttcta ggtgccaggccctggagacacatcagtgaacaaaacagacatcatcccacctctttccactacag gccaagcaccatgctggtctctgggaaccctgttgtgagcaagacagacccaggcttacccttgt ggactcatgttacaggcagggagacgggcacaaaacacaaataaaaagcttccatgctgtcaga agcactatgcaaaaagcaagatgctgaggtactgctaagctgtgtgggatgggggctcagcccg gccagggaccaatgatggagaccggggcggggtccagaggtaacttgtgccccgccccgata gcctagggcccagcggggattaagttttttttttttttcttttgaggaatcttactccaatgatggcgcc tgctcaattcgaggaatcttactagaccggggcggggccacataaggttgcaaatcccacagag agcctgcgcctggggttgttttctgtaggcgcctgtgcccccgcccctgccaccaatgatggcgc ctgctcaattcctatgtgtctggcaccagaaacggaagctgcaggttgcagcccctgccctcatggagctcctcctgtcagaggagtgtggggactggatgactccagaggtaacttgtgggggaacgaSEQ Promoter SequenceID NO:acaggtaaggggctgtgtgacgagatgagagactgggagaataaaccagaaagtctctagctgt ccagaggacatagcacagaggcccatggtccctatttcaaacccaggccaccagactgagctg ggaccttgggacagacaagtcatgcagaagttaggggaccttctcctcccttttcctggatactga gtacctctcctccctgacctcaggcttcctcctagtgtcaccttggcccctcttagaagccaattagg ccctcagtttctgcagcggggattaatatgattatgaacacccccaatctcccagatgctgattcag ccaggagcttaggagggggaggtcactttataagggtctgggggggtcagaacccctagttcat ccagctggagcgagccgaacggtcgttgagccaagaggtaagggtttaagggatggttggttg gtggggtattaatgtttaattacctggagcacctgcctgaaatcactttttttcaggttggccgcggat ccgccacc24 V14 (Enh2- ctataatcctggcactttgggaggccgaggagggtggatcgcttgagccaggagttcaggacca TFBS- gcctgggcaacatagcgagactccacccctacaaaaaatacaaaaactagtggtgtgcacttgtg Rho(BTIm gtcccagctactcaggaggctgaggtgagaggatcgcttgagcccaggaggcagaggctaca d2)-MVMi) gtgagctatgattgtggcactgcactccagcctgggcgacagagaccttgtctcaaaacttttttttt cttcgtcaagctttacagaataaagagcactgtcacctcagtgatggctgttagttccccatcacca gggctccatgaggttgcaattgtgaaactcacaaaggaggaacctgagacagagaggggaagt actgagatcatctaggtccattcccccactcactcgttcattcaacaaatattcaggagcaccttcta ggtgccaggccctggagacacatcagtgaacaaaacagacatcatcccacctctttccactacag gccaagcaccatgctggtctctgggaaccctgttgtgagcaagacagacccaggcttacccttgt ggactcatgttacaggcagggagacgggcacaaaacacaaataaaaagcttccatgctgtcaga agcactatgcaaaaagcaagatgctgaggtactgctaagctgtgtgggatgggggctcagcccg gccagggaccaatgatggagaccggggcggggtccagaggtaacttgtgccccgccccgata gcctagggcccagcggggattaagttttttttttttttcttttgaggaatcttactccaatgatggcgcc tgctcaattcgaggaatcttactagaccggggcggggccacataaggttgcaaatcccacagag agcctgcgcctggggttgttttctgtaggcgcctgtgcccccgcccctgccaccaatgatggcgc ctgctcaattcctatgtgtctggcaccagaaacggaagctgcaggttgcagcccctgccctcatg gagctcctcctgtcagaggagtgtggggactggatgactccagaggtaacttgtgggggaacga acaggtaaggggctgtgtgacgagatgagagactgggagaataaaccagaaagtctctagctgt ccagaggacatagcacagaggcccatggtccctatttcaaacccaggccaccagactgagctg ggaccttgggacagacaagtcatgcagaagttaggggaccttctcctcccttttcctggatactga gtacctctcctccctgacctcaggcttcctcctagtgtcaccttggcccctcttagaagccaattagg ccctcagtttctgcagcggggattaatatgattatgaacacccccaatctcccagatgctgattcag ccaggagcttaggaggggcggcgcctatataaggtgtgggcgggtgtggcacagctagttccg tcgcagccggcgagccgaacggtcgttgagccaagaggtaagggtttaagggatggttggttg gtggggtattaatgtttaattacctggagcacctgcctgaaatcactttttttcaggttggccgcggat ccgccacc25 VI 5 (Enh2- ctataatcctggcactttgggaggccgaggagggtggatcgcttgagccaggagttcaggacca RHOpatent gcctgggcaacatagcgagactccacccctacaaaaaatacaaaaactagtggtgtgcacttgtg (MD)- gtcccagctactcaggaggctgaggtgagaggatcgcttgagcccaggaggcagaggctaca MVMi) gtgagctatgattgtggcactgcactccagcctgggcgacagagaccttgtctcaaaacttttttttt cttcgtcaagctttacagaataaagagcactgtcacctcagtgatggctgttagttccccatcacca gggctccatgaggttgcaattgtgaaactcacaaaggaggaacctgagacagagaggggaagt actgagatcatctaggtccattcccccactcactcgttcattcaacaaatattcaggagcaccttcta ggtgccaggccctggagacacatcagtgaacaaaacagacatcatcccacctctttccactacag gccaagcaccatgctggtctctgggaaccctgttgtgagcaagacagacccaggcttacccttgt ggactcatgttacaggcagggagacgggcacaaaacacaaataaaaagcttccatgctgtcaga agcactatgcaaaaagcaagatgctgaggtactgctaagctgtgtgggatgggggctcagcccggccagggaaaaccagaaagtctctagctgtccagaggacatagcacagaggcccatggtccctSEQ Promoter SequenceID NO:atttcaaacccaggccaccagactgagctgggaccttgggacagacaagtcatgcagaagttag gggaccttctcctcccttttcctggatactgagtacctctcctccctgacctcaggcttcctcctagtg tcaccttggcccctcttagaagccaattaggccctcagtttctgcagcggggattaatatgattatga acacccccaatctcccagatgctgattcagccaggagcttaggagggggaggtcactttataagg gtctgggggggtcagaacccctagttcatccagctggagcgagccgaacggtcgttgagccaa gaggtaagggtttaagggatggttggttggtggggtattaatgtttaattacctggagcacctgcct gaaatcactttttttcaggttggccgcggatccgccacc26 VI 6 (hRK- tgtagttaatgattaacccgccatgctacttatctacgtacatttatattggctcatgtccaacattacc TFBSRho) gccatgttgacattgattattgactagaattcgctagcaagatccaagctcagatctcgatcgagttg ggccccagaagcctggtggttgtttgtccttctcaggggaaaagtgaggcggccccttggagga aggggccgggcagaatgatctaatcggattccaagcagctcaggggattgtctttttctagcacct tcttgccactcctaagcgtcctccgtgaccccggctgggatttagcctggtgctgtgtcagccccg gtctcccaggggcttcccagtggtccccaggaaccctcgacagggcccggtctctctcgtccag caagggcagggacgggccacaggccaagggccctcgatcgaggaactgaaaaaccagaaa gttaactggtaagtttagtagaccggggcggggtccagaggtaacttgtgccccgccccgatag cctagggcccagcggggattaagttttttttttttttcttttgaggaatcttactccaatgatggcgcct gctcaattcgaggaatcttactagaccggggcggggccacataaggttgcaaatcccacagaga gcctgcgcctggggttgttttctgtaggcgcctgtgcccccgcccctgccaccaatgatggcgcc tgctcaattcctatgtgtctggcaccagaaacggaagctgcaggttgcagcccctgccctcatgg agctcctcctgtcagaggagtgtggggactggatgactccagaggtaacttgtgggggaacgaa cgcgtaaggggctgtgtgacgagatgagagactgggagaataaaccagaaagtctctagctgtc cagaggacatagcacagaggcccatggtccctatttcaaacccaggccaccagactgagctgg gaccttgggacagacaagtcatgcagaagttaggggaccttctcctcccttttcctggatactgagt acctctcctccctgacctcaggcttcctcctagtgtcaccttggcccctcttagaagccaattaggc cctcagtttctgcagcggggattaatatgattatgaacacccccaatctcccagatgctgattcagc caggagcttaggagggggaggtcactttataagggtctgggggggtcagaacccagagtcatc cagctggagccctgagtggctgagctcaggccttcgcagcattcttgggtgggagcagccacg ggtcagccacaagggccacagccctttttgtcttttatttcaggtcccggatccggtggtggtgcaa atcaaagaactgctcctcagtggattttgcctttacttctaggcctgtacggaagtgttacttctgctctaaaagctgcggaattgtacccgcggccgcTable 4. Illustrative components of expression constructs disclosed herein.SEQ ID NO: Component name Sequence11 WPRE6 aatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactatgtt gctccttttacgctatgtggatacgctgctttaatgcctttgtatcatgctattgcttc ccgtatggctttcattttctcctccttgtataaatcctggttgctgtctctttatgagg agttgtggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacg caacccccactggttggggcattgccaccacctgtcagctcctttccgggactt tcgctttccccctccctattgccacggcggaactcatcgccgcctgccttgccc gctgctggacaggggctcggctgttgggcactgacaattccgtggtgttgtcg gggaaatcatcgtcctttccttggctgctcgcctgtgttgccacctggattctgcg cgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttccttcc cgcggcctgctgccggctctgcggcctcttccgcgtcttcgccttcgccctcag acgagtcggatctccctttgggccgcctccccgc12 BGHpA ctgtgccttctagttgccagccatctgttgtttgcccctcccccgtgccttccttgaccctggaaggtgccactcccactgtcctttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattctggggggtggggtggggcaggaca gcaagggggaggattgggaagagaatagcaggcatgctgggga 38 WPRE3 aatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactatgtt gctccttttacgctatgtggatacgctgctttaatgcctttgtatcatgctattgcttc ccgtatggctttcattttctcctccttgtataaatcctggttagttcttgccacggcg gaactcatcgccgcctgccttgcccgctgctggacaggggctcggctgttgg gcactgacaattccgtggtgtt39 WPRE250 aatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactatgtt gctccttttacgctatgtggatacgctgctttaatgcctttgtatcatgctattgcttc ccgtatggctttcattttctcctccttgtataaatcctggttgctgtctctttatgagg agttgtggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacg caacccccactggttggggca

[0234] Immunohistochemical analysis of eGFP expression in hiPSC retinal organoids

[0235] Analysis of eGFP expression and cell specificity in ROs was completed was completed using a standard protocol known in the art to fix, embed, cryosection and stain retinal organoids (RO) derived from induced pluripotent stem cells (IPSCs) and human embryonic stem cells (HESCs). Primary and secondary antibodies used are listed in Table 2.

[0236] Subretinal administration of AAV to test novel promoters in wild type mouse retina

[0237] To assess promoter activity in the murine retina, adult WT C57BL / 6 mice received two subretinal injections (each 2 pl) of AAV2 / 5 vector expressing eGFP under the control of different rod-specific promoters disclosed herein at a final dose of 4E9 VG / eye. A list of all promoters and respective AAV2 / 5 vectors is provided in Table 1. Subretinal injections were performed as known in the art. For each vector, four injected eyes were used for qPCR expression analysis (n = 4 eyes) and immunohistochemistry analysis (n = 3-4 eyes). To serve as controls, three eyes were left uninjected (n = 3 eyes) and two eyes were injected with PBS (n = 2 eyes).

[0238] qPCR analysis of eGFP expression in mouse retina

[0239] Four weeks post-injection (pi), retinas were harvested and frozen on dry ice as known in the art. RNA was extracted and cDNA was synthesized at 5 ng / pl. Absolute eGFP gene expression levels were measured using a qPCR assay targeting the eGFP sequence with 50 ng per reaction.

[0240] Immunohistochemical analysis of eGFP expression in mouse eyes

[0241] Four weeks post vector administration, whole eyes were enucleated, fixed in 4% PFA for 3 h, embedded in OCT and sectioned. Sections (18 pm) were stained with Peanut Agglutinin (PNA), which selectively labels cone photoreceptor cells. Details of primary and secondary antibodies used are listed in Table 2.

[0242] Example 2. Promoter activity in hiPSC retinal organoids

[0243] Wild type human iPSC-derived retinal organoids (ROs) were transduced with AAV2 / 7m8 vectors expressing eGFP at a dose of 1E11 VG (viral genomes) per organoid between differentiation day 130 to 170. Three weeks post-transduction, ROs were analyzed for eGFP expression (FIGS. 2A and 2B); organoids were dissociated into single cells and stained with CD73 antibodies to enable identification of rod and cone photoreceptor cells via flow cytometry (FIG. 2C).

[0244] Median fluorescence intensity (MFI) was used to quantify eGFP protein expression levels in transduced photoreceptor cells (FIG.3A). The median fluorescence intensity in eGFP positive / CD73-APC positive photoreceptors (between 20-61% of CD73 positive cells), was normalized to expression in organoids transduced with RK-eGFP and expressed as a fold change relative to RK (FIG. 3B).

[0245] In human retinal organoids, five promoters, 3, 4, 5, 7 and 9, showed an increase in GFP intensity relative to the RK baseline.

[0246] Example 3. Immunohistochemical analysis of eGFP expression in hiPSC retinal organoids

[0247] The cell specificity of each promoter was determined by immunohistochemistry. ROs were harvested 3 weeks post-transduction, fixed, embedded and cryosectioned. Cryosections were stained to identify vector derived eGFP protein and cone photoreceptors through the identification of L / M opsin immunoreactive cells. Analysis of RO sections via confocal microscopy demonstrated significantly increased eGFP signal for vectors 3, 4, 5, 6, 7 and 9 in rod photoreceptors when compared to RK eGFP expression levels (FIGS. 4A-4H).Qualitative comparison of eGFP signal to L / M opsin expression demonstrated RK promoter activity in rods and cones, (FIG. 4 A), consistent with the known pan-photoreceptor activity of this promoter. Conversely, no significant overlap was detected between eGFP and L / M opsin in retinal organoids transduced with vectors 1, 3, 4, 5, 6, 7 and 9 (FIGS. 4B-4H), indicating rod photoreceptor specific promoter activity.

[0248] Example 4. Promoter activity in murine retina

[0249] qPCR analysis of eGFP expression in mouse retina

[0250] Frozen retina samples from mice subretinally injected with AAV2 / 5 vectors expressing eGFP were analyzed using qPCR (FIG. 5). eGFP expression levels for each vectorwas normalized to eGFP expression levels driven by AAV2 / 5 containing an RK promoter. Promoters 1, 3, 6 and 9 significantly exceeded RK expression levels (best performers also in the context of organoid data: 3 and 9 indicated by black columns).

[0251] Immunohistochemical analysis of eGFP expression in mouse eyes

[0252] The cell specificity of each promoter was determined by immunohistochemistry. Four weeks post injection, whole eyes were harvested, fixed, embedded and cryosectioned. Sections were stained with PNA, a cone photoreceptor-specific marker, to confirm that the eGFP fluorescent signal was specific to rod photoreceptors.

[0253] Analysis of retinal sections via confocal microscopy demonstrated significantly higher eGFP signal for promoters 1, 3, 4, 6 and 9 in rod photoreceptors than RK. Representative images are shown in FIGS. 6A-6F. Of note, promoter 4 showed some un-specific transduction in the RPE (arrowhead). Promoters 1, 3, 6 and 9 showed highly specific rod transduction as indicated by the lack of colocalization with PNA. Images of promoters 2, 7, 8 and 10 are not shown due to their low levels of expression either in this or the organoid platform (FIG. 5).

[0254] In sum, in human retinal organoids, five promoters (3, 4, 5, 7 and 9) showed a marked increase compared to the RK baseline in the intensity of eGFP fluorescence and a strong rod-specific tropism (FIGS. 3A, 3B, and 4A-4H). In the murine retina, four promoters (1, 3, 6 and 9) showed a marked increase compared to the RK baseline in the expression levels and a strong rod-specific tropism (FIGS. 6A-6F). Promoters 3 and 9 provided high levels of specific expression in both mouse and human rod photoreceptors. The differential expression profile of certain candidates highlights the need to perform promoter screens in experimental platforms from multiple species. High performance in both is useful for achieving translation of treatments from preclinical to clinical setting.

[0255] Access to a potent rod-specific gene therapy toolkit allows not only for treatment of defective rods but also could prolong survival of cones that otherwise degenerate due to a bystander effect. In addition, optogenetic intervention in rod cells aiming to selectively increase their sensitivity and effectively act in a cone-like manner could be developed as a therapeutic strategy for patients with macular degenerations.

[0256] Example 5. Second generation rod promoters

[0257] In this study, a second generation of promoters were rationally designed by combining elements from those promoters that drove strong and highly specific expression in human retinal organoids and mouse retina from the first generation of rod-specific promoters.The second-generation promoter sequences that were used in this study are summarized in Table 5 (see also Table 3 for sequences as well as Table 6).Table 5. Summary of second-generation promoters.SEQ ID NO: Promoter number Promoter name22 V12 Enh 1 -TFB S-Rho(MD)-MVMi 23 V13 Enh2-TFB S-Rho(MD)-MVMi 24 V14 Enh2-TFB S-Rho(BTImd2)-MVMi 25 V15 Enh2-RHOpatent(MD)-MVMi26 V16 hRK-TFBSRhoTable 6. Promoters and promoter componentsSEQ Promoter Comment Comprises (from 5’ to 3’)ID #NO:1 1 SEQ ID NO: 13 comprises SEQ ID NO :1SEQ ID NO :15 2 SEQ ID NO: 15 comprises SEQ ID NO: 5SEQ ID NO: 510 3 C + SEQ ID NO:20 + SEQ ID NO:33 +SEQ ID NO: 17 + SEQ ID NO:34 4 4 SEQ ID NO 35 + SEQ ID NO:21+CGCGGCCGC3 5 or V5 SEQ ID NO:36 + SEQ ID NO:21+CGCGGCCGC8 6 SEQ ID NO: 18 + SEQ ID NO: 17 6 7 SEQ ID NO: 16 comprises SEQ ID NO:33 + SEQ ID NO: 17 + SEQ SEQ ID NO: 6 ID NO:347 8 SEQ ID NO: 16 comprisesSEQ ID NO: 79 9 SEQ ID NO: 19 + SEQ ID NO: 17 2 10 SEQ ID NO: 14 comprises SEQ ID NO:37 + SEQ ID NO:21SEQ IDNO:222 V12 SEQ ID NO: 18 + SEQ ID NO: 27 + SEQ ID NO: 28, + SEQ ID NO: 2923 V13 SEQ ID NO: 19 + SEQ ID NO: 27 + SEQ ID NO: 28 + SEQ ID NO: 2924 V14 SEQ ID NO: 19 + SEQ ID NO: 28 +SEQ ID NO: 27 + SEQ ID NO: 30 25 V15 SEQ ID NO: 19 + SEQ ID NO: 28 + SEQ ID NO: 2926 V16 SEQ ID NO: 31 + SEQ ID NO: 28 +SEQ ID NO: 32

[0258] Promoter sequences from second generation were designed, cloned into an AAV backbone carrying eGFP, and packaged into AAV vectors with AAV5 or AAV7m8 capsids. As controls, vectors were generated comprising the commonly used rhodopsin kinase (RK or GRK1) promoter driving eGFP expression.

[0259] To assess promoter activity, AAV7m8 vectors were used to transfect human induced pluripotent stem cell hiPSC derived retinal organoids. Three weeks post transduction, the organoids were dissociated into single cells for flow cytometry analysis or for immunohi stochemi stry .

[0260] As seen in FIG. 8, first generation promoters increased median GFP intensity in retinal organoid photoreceptor cells in this experiment by 2-3 fold. In contrast, second generation promoters V12-16 increase reporter gene expression 5-9 fold. Notably, rod promoters resulted in rod photoreceptor-specific GFP expression (FIG. 9). Consistent with this, V12-V16 did not drive expression in cone photoreceptor cells (FIG. 10).

[0261] In parallel, promoter activity was assessed in murine retina. Wild type mice received subretinal injections with the AAV5 vectors. Four weeks post vector administration, eyes were harvested for immunohistochemical analysis and qPCR expression analysis to determine specificity and expression levels, respectively.

[0262] qPCR analysis determined that V5 andV12-V16 increased GFP expression in murine retina by up to 28-fold in average, with V5 and VI 6 showing a particularly strong increase (28- and 20-fold, respectively) as compared to the RK reference promoter (FIG. 11).Further, gene expression driven by the V5 andV12-V16 promoters was specific for expression in rod cells, as evidenced by the fact that GFP was not expressed in cone photoreceptor cells of mouse retina frozen sections (FIG. 12).

[0263] In sum, in the murine retina, particularly V5, V12, V14, V15, and V16 showed a marked increase (7-28-fold change) compared to the RK baseline in reporter gene expression levels and a strong rod-specific tropism (FIGS. 11 and 12). In human retinal organoids V5, V12, V13, V14, V15, and V16 showed a marked increase (6-9 fold) compared to the RK baseline in reporter gene expression levels and a strong rod-specific tropism (FIGS. 3, 8 and 10).

[0264] Successful preclinical development of AAV vectors requires the use of promoters, which are active in both human and small animal models in order to carry out meaningful assessments of both safety and efficacy. The data presented demonstrates that target gene expression levels from the commonly employed RK promoter can be improved by rational design incorporating data from single cell RNA sequencing and other sources.

[0265] Additional sequences

[0266] SEQ IDNO 13 ccctggaaccctcagggaagagggcactgcactgtctgccagcccttgagtctccaggagtggagattagctgtggccagtatccag ggtaagcgtgaggggatcagggagagtggtggactcagcaggccagggattagcagatgatactccaccctcagcccctgcagga ctgcaggctcccagaaaagccaggctgtgagtgcccaaggccagcaccccaaagccctggggcctgcccaccatcctggtcacaa ccattagagacagacccccaaggtgaggaaaccccaggagatcctaacttatttgcatatattggcatgtgcttttgcatatacctgccca ggggctgaacctgatgggtcttggagcccctcctgtgaaattcttgtaggatttgtttccccagttgctcctcctccagtgctccatacagc ttccagaacagcattcctaactgctaccccttacaaaaaaccttcccagagttgactccactcaccaccccacccccaatttatttcttgac tcttttaaagttataaaatagaagaaatacagaaagtgaataaatatctaatgtaactaataatttacaaaatgagaccccgtataactacca cccaccctggtcaagacatggagtactacttttaaaacagtttatcagtttcttaaaaaatgaaacagaaatgtaccatatgattgggtcat ctggaagattctaccccaggaggcttcccaagagaaatgaaagtatattgtgaggcacgatggctcacacctgtaatcccagcactttg ggaggctgaggtgagcagatcacttgaggttgggagtttgagaccagcctggccaacatggtgaaaacctgtctctactgaaaacaca aaaattagctgggcgtggtggtgcaagcctgtaatcccagctactcaggacgctgaggcaggagaatcgcttgaacctgggaggcg gaggttgcagtgagccgagatcgcagcatcgcactccagtctgggcggcaagagcaagacttcgtctcaaaaaaaaaaaagtatata tcttcacaaagacttgcacacaaatgttcatagcaacatgatgtatcgccaaaaagtggaaacaacctgtgtccactgatgagtggataa actgtggtgcgtctataccatgcaatactcttgggcaatgaaaagaaatgaagttctgatacaagctacgaggtggatgaacctcataaa acattacgctacatcaaagaagccagatgtgaaaactcacatattgtatgacccatttatataaaatgtccaggaaagacaaatctataga cgcagaaagcagatgtgtggttgcctgggactggggtgtacgaatgaagagtggttatcatctggtgtgaggggtcttactggggcaa cgggaatgttctaaaaccagatgtggcaagagttgcacagttcagtcaatgtgttaaaaatcatgagttaggccggggcggtggctcac gcctgtaattccagccctttgggaaccccgagtgggtggattgcttgagcccgggagttcgagaccagcctgggaaacatggcgaaa ccctgtctctacaaaaaatatgaaaattaaatgggcatggtggcatgtgcctgtggtcacagctacttgggaggctgaggtgggataatc acttcagctcagaaggtcgaggctgcggtgagctatgatcacgccactgcactccagcctgggtgatagagtgacaccctgtcttaaat aaataaataaataataagtaatataaaagtcattgagttgtatacttaccatgggtgaattttatggcatatatattcccataagcttgttttgaa aaatagatgccagccaagttagtaaaagaaagcaagacattggccctgccagtccctagaagcttccacatacaccttcccaattacaa cccccaacccagcaagaaggaaccactctcccatcttgtagtacaacctgtgcttttatttatgggcaactatggaaaactttatttaacaa ctctaggctttatagaaaactagaggtggcaaatgtacttgtgtttgtagccatagacaggacagactaattttgccttttgggaattttgtat gcatggatcactggggtcttcttttgaatctggcttctttctctccatatatatgcacatgagtttcatccaaactttagccactgtgtgtggct gtgttttgctcattttccccaccctagagcccactactgttccctcaggctcatctaattttccctccttcccactctgcaggggctccaactg cctctaagttgcccaatctcaggacgcatttcaagtctcagctcagacagtggccccttggatcattccctcaccccaacacagccacta ctcttggctttcacctcaccccatgcttctgcttctccatccccatccctggcagctccttcaggctcctttaccagctcctgctctttttgcat cctctgagtgagcccgtgttccaggtgccacctctggtcatctttgtttccttgggtttccgaggtgatatcagtcagtcctgtggctttaag catcatctgcaagctgaggcctccaaaatttctatctcctgtagacctctccctaagtctcacacacacacacacacacacacacacaca cacacacatttcaacctggatgtctctcagacttttttttttttttgcttaggctggagtgcagtggtgtgatcttggctcactgcaacatccac ccaccgggttcaggcaattctcctgcctcagcctccagagcagctgggattataggcacctgccaccacacccgactaatttttgtatttt tagtagagacggagttttgccatgttggccaggctggtcttgaactcctgacctcaggtgatccacctgctttggcctcccaaagtgcta ggattacaggcgtgagccactgcacccagcctctctcagacatcttaacaaagtagtgctcatgattttccctccaaaattcttcctccaa acttccagcctcaggcttcttcttttcagtacatgtgtaaaccagcctggggcggtttactggttcacccactaaaccagctacccagtttc ccagggtgagcccctcaatgcccgcctttttcctcccctttcccttctatctgtactcccaccagcatagtccaagccccatcatccctccc gtcaggctgcaggcaagggcgaatgtcacctgtgttagtctcaggcccaagcagagcagaagggcttgctgctgttccttggggagg agctctgacccctgcagcctggcatatttgtagggaagggtcagggagtacattgagctttaccgttggggtctggccttgctggcaga aagaactttcttgtgaatcacttgccatgtagaggttgcattatcagaggtcttgaagcccaacactggaggggaagccaggggtcatc cagtcaccaatcagcttgtccctccgcagcctcctgaccagtttcctgtttccatgcttgcctgctacacagagccacacagatcatgccc agcccctgttcaaaaccatcatctaaaactacactccttcctgtggcttacagtctcacaacatctgcccctgccctctgtcaacctcatct cccaccactctcccctgcacccgttaagctccaggctccccggttcccattctgctcctcaaggctcattccagcctcagaacctttctcg ttgcagtgtccttggcctggagcaccactcacccagcttttttcacagctgagtctttcttacttttcatgtctcagttcaaatatacatccacc tcgtcataggggcctgcctacctgcccctactgccctccacaacatgcttctgccctgtttgtttcttttatggcacttagcataatttgcaat attctttatttgtcagtaatctcctccattggaatgtaagctccatgaggacagggcctatgtctgtcttgctcagtgtgccaaggctggcac actgccaggcacaaaggaagagctctaggccaggtgcaggggcttacatctgtaatcccaatactatgagaggctgaggcaggaggattgcttgcatccaggagtttgagaggttgaggctgcaatgagctgtgattgcaccacttcacttagcctgagtgacagagagagactgt ggcttaaaaaaaaaaaaaaaaaaaggaaaggaaaggaaaggaagaactctagaaatggtgcggataattaagtcaagtcctgccctt tgccaaatatccctcagggccaccacactgagcccctcttccttgggccctgtcccttgcaccctctcctcctagatctctgtatcccttca agtgtccagcctcagttcacagggcaggtcagatgcctcagtcatctgccgggtcttggatccatcctcaaggcccaggcctgggaca cagcctttctggcagagcccaaggcagggctgctgagaccctgcccagagaagactctaagccagaaagaacctgcctgcgtcctg gcctgggggctggagagttggggtacacctcttgccaccgctgagtgggagctgacaagcccctgctaccatgaggagtctccagg catcacatgggaactccaagctgttattctcccatcagccactccgtgtcttggccaaggtgtatgggaagccaggcccctggtaagcc gggcaggtcccctggcaacccatggtgctccaaacctgggccatgcaggagcccttccctggggcccacatccttgctggaccctgg gagttgggaggtgctgctggtttcctgggcatcaacagcctcttttctccccctcccccattcagcacctgctaatcctttgcacacggcc cagctactgccaattaatcttccttgtgtctgaggggccagctacgagtggcagcaagaaggcaattcctggctggcggttggcatcta agcaggtaactcagggtgaggtggaggctcggtgggaccccgaggggatccccatgttctccctctagaaaggcatttaggtaggac caaactggcacggtgggaaacctgtggcttccctctatctggtggagcttggactttagcgggcatcagaattttcaggacagcttgtac aatgcaaattcctaggtcccccaggaggacttgggggacctaggaatctatattttcactagctctccagaagactctggaaatgctggt ggtgcaacaccatggctccccgagctgccactcgtgggtcccccttcccacctgagcctccctcatcaaagtgcagtcagcagggac acgggtgaaagatgggagggacctgcaaaaattctcatttaggaggaaaaagaaaaatgggggatcggggaggcttccttagtcattt ccagaaggtttagaaatgcaagcgagaattgctgcctgctgggaagggtctacaggcctggcccgcggaagaggccaacactcgg caaatgccacgactacctgggaatacatgtggggcagctggaatcgggcagacctgagtgtaaactcaagttggccactgccggctg tgctgcttcaggcgattctgttgacctttctgtgctacataatgaaaattcttattttactgagtgcaggggctgggcggggcgtggtggtg agagcgaggatgagtatgatcatgggtagaatgcacttagctcagccctggctctcggggctgcctggggctggctggggtcagtga aatctgctatcccaacatgtgagctctggcagtcccgacctgggaaaacattgtgtctgtgcccccaccttcagggaaatggaagacca gaggggtcatgtctggcccaagttcattttttgggaggctgagccagggatctgacacttagagctgtgttcccaccacagcctctcgtg ggagaggctgggctgctgatgccagagtgggaacagaggtcagggctgtaagtagaggacaggcagcacagccgtgggctctgg ggagctgggcttgggggcaaggatcctgggcctgggctgccctcttcagctggggccagggtccctggcagaaaggggcatacag actctctgccaaagcccgggttgcccatggctgggagaggggtggggcacactcatgtggggcaagaacacatctagggctagga gttagcacacactgacctgcacgctgcgcactggccaggccatcctgcaggggttcagtcataagggctggggctccccattcccca aggagaaatcaaccagaaacaacctgtacttaaggtaggactgaaataacacagtatatcatgaaaaaagtcttacctggcattttatctt tttctttctttctttttttaaaatatagcactataagcattttatttcagttttgagacagggtcttactctgtcgaccaggctggagtgcaatggc tgagtgcaatctccaccttttgggctccagtgattctcgtgcctcagcctcccgagtaactggtactacaggcacgtgccaccacgctgg gctaattttttgtatttttggtagagacagggtttcatcatgttgcccaggctggtctcgaactcctgagctcaagcgatctgcctgcctcag cctcctgaagtgctgggattgcaggtgtgagccacaccatgcccagcctttttgtttgtctgtttgtttttgagaaggagtcttgctctgtca cccaggcaggctggagtgcagtggcacgatctcggctcactgcaacctctgcctcccgggttcaagtgattctcccacctcagcctcc ggagtagctgggattacaagtgcacaccaccacacccagctaatttttgtatttttagtggagatggggtttcaccatgttggctaggctg atctcgaactcctgacctctggtgatctgcctgccttcacctcccaaagtgctgggattacaggcgtgagctaccacaacaggcccattt ttttttctttttttgagacaggatcttgctttgttacccaggcagaagtgcagtggcatgatcactgaagcctcgaccccttgcctcagcctc ctgagtagctgggactacaggtgcacaccaccataacaggctttttttttttttttttttaaatagagacagggtcttgatatgttgcccacag tggtctcgaactcctgagctcaagtgatcctcccatcttggcctcccaaagtgctgggatcacaggcatgagcctctgcacccagccca ttttatcttaaaaattaaaaaatgatgaacataggttaagtttatcttaagacagattttccactcatgtaattactcataatgaatatttttactta taatgagattatgtagtacatgcatattacacaattagtgttttcatttgataccatgtaatagtcatagtcaacatttattaagccctcggcat gtgccagtcactgttctgagtatgttacatttattattattattattatttttgagacaggatctcactctgtcacccaggctggagtgcagtgg cacgatcttggttcactgcaacctctacctcctgggttcaagagattctcatgcctcagcctcctgagtagctgggattacaggtgcctgc aaccatgcccagctaatttttgtatttttagtagacaaggggcttccccatgttgccaggctgctctggatctcctgaccacaagtgatcca cccacctcagcctcccaaagtgctgggattataggtgtgagccactgcccttggcctgagtatgttacacttatcaattcattttatcctcac aattagcccatgaggtaggtactattaatatccccattttacagacaagcaaactgaggcacagagacattaactgatctgctcatgactt gacttcagagcaggcacatcttcttgctagctcaattagactgcttctttctttttaatgcccccataatattctattattgggtatagtgtaactt atttaatacatgcccctcttagagatgaacatttagttacatgtaggtactttccaataaccagtctcctcttgatgaatatttgggtgtttttcat cttttgctataacattgttgcaaccagcttctttgtacacatatatttatggccttgagtaattctataaggtaaatacctagagatataattgca gaaatatatacacacacacatatctgtgtgtatatatgtaatatgtatgtgtatatgtatatcaaatatatatatgtatgtgtatatatattatatat tttttgagacagggtctcactctgttgtgcaggctggagtacagtggtgcatcatgacccactgctgcctcgacctcctaggctcaaacg atcctcccacctcagcctcctgagtagctgggactacaggagtgagccactacacctggctaatttaaaaaatattttgtagagacgagg gtctcgctgtcttgcccaggctgatctcaaactcctggattcaagcaatcggcccatctcagcctcccaaattgctgggattacaggtgtgagccaccgcacccagtcgaaatgtttttaaagttgatgaaggaaaaggaaatgtacaactcataagagtggcccataagtggcgggag ctactgtgagtgctccagggagggctaaggactctgtctcctacaggcatcaggatgttgggctgggtccagagggtgctgcctcagc ccccagggacccctcggaagaccaagatgcaggaggaagaggaagtggaaccagagccagagatggaggcggaggtggaacc agaaccgaatcctgaggaggccgagacagagtccgagtccatggtgagaggtagagcgggaggaggggcaggccggcctcgct gctcagggtcttcccagtttgggccatctgctctccagggactttctacagcagaaaaggttggcatcagaaggaaaagtgggggtca cagggtcccacacttgttcgagtgagaaaactcaggttcagacagtgggtagggcttgcccaaggctaacattttctgtaaaaggatag atagtaaatattttaggctttgtgagtgatgctatctctgctaattactcaactctgccattatagtgtgaaaacagttataggtaatgcatgaa tgaatgagcatagctgggttccaataaaattttatttataaaaacaagcaatagcctagatttggcccacaggtcgtattttgccaacctcta gagttcagaacagagctactcacaggagccagtccatgaactgtttattactagtccatgatgattaaacagtttgtaccagaatgtaaat ctactctgtagctaagtacactgattagtttagttgctatttattttgtaccaagactttctcaatgaaggaagcagtgcattgataaacattct ggtgcaaactgcttaatcatcatggaccagtaacagtttgcagactggcacagatccatggaccacactgtggagtatttctccagaact tttcctctgcaacccatccattcaggagctggatgacaactaattcagcagctcaattcgacacatttgatttagcttctaattgatgccagg ccctgtggtcagtgctgaaagcataaagatgaataggcattgccccggggactcagcagacaagagcccagccagttcattctgcca agagacataggtggagaggagggaacgtggtgagtctcaacattggcctcaactctccccttcttggggagccaaataagtgtgtcag accaaactatgggagctcagaagagggagagtcatcttaactttggagattggaggctgatgggggagagcttcttagacgcagagg gtggagcagaattttaactgagggagatggagaggctgggcattcctggcagaggaaaggctgttactagagaagggcagggtgtg tttaggaaacaggtatagggtgtgcgtatgcatgtgtatatgttgcatgtatatacacgtatatgtacatatatgtgtgtagtgtgcccttggt atgtctgtgtgtgcacacatgtatttgcacaatgggtatacatgacaatgtatgtgaacatgtatatgtacatgaatgcatgtatatgtgtttg cacatatgtttatgtgcacatgtattagtatgtctgtggttccatgtgcttgtgtgacatgctagtgtgtatgtgtgtgtatgtgtctacatatttc tgagtatgtgagtgtactcatgtatttctatgtatctgcatgtttgtgcatctgtgaatgtgtatatacacatatttgcctgtctccacatgtgtat atatacacattttgtgtctgtgcatggatgttcatgtatatttaggtgcagaaatctgtgtgtatatgcatgtccatatggacatgcttctgtgta tgtgcaaatatgcacatgtgtgtgactgagcatatgtgtgtatactcatatttctgcacctgtatacatctgcaatgtacctatgcatatgtgc atgttgtgtgtctgtacacaggcagttatacgcatgtatgtatctgtatgaatgcatgtgtgtatatgcacgtttgcatatgactgtatgcaca tatatgtgttggggcatgcatgcatatgtacatacacatgtatatgtacatatgtttttgcctgtacgtgtgtgcacatgcacttatttttgtgtct gtgcatgtgtgtgtatatatgcatgttggtatttgtgtgtcattgtatacatgcatatgtatgtgtatatatgctcacattgtatgtctgttcatgta tgtatgtgcatgagagtgagtatgcacacatttatgtgtctgtgtatgtatgtgcatgtatatttgtttgtacatacacataagtgcacgtagta gcaggaggtgtggctgcagaggcaggtgaaaggcagatggcagagaccaaactccacctagctactagggaggaatggaaagttt gggccagaaggagagggagagcactggcagaggatgaggggcgtgggctcctctcacctccctcttcccctcagccagccccaag gcttcataactatctctaaatggtcctttctcctctcaggccactgaagtggggcagcctcctggggatgttgtaggcctgggaggcttgg aggccaccttggaccttgggggctgtgagggcccccagccccagatgaggggtccctggggctaggaggaagggagtatcccag gtggaaggtaaggccaacaggatttacagagatgatcctgaggaaagagtcgttcaggtcttttgtggggcagggatcccttgaggga gaatcaggctgcccctgtccaaaccaggctggtctcacctaggacttacatgatcctgttcgaaccctctagccccccgaagagtcatt caaggaggaggaagtggctgtggcagacccaagccctcagggtgagtgcgagtgctgaagcccaggctggcatggcagggggg cccgcattctcccctcggagccgcagagcccacgaggctggcggacccatttcacagatgaggaaactgagactcagagaagctaa gggtgggctcccagagacttcagttgagctgagctggagcctggctttctgacccctgggggtctctctcctctcccctgacttgatctc cccaagcagggtagaaatgggcctgtctgaggccctagatctgaaggatgagagattcttttccagagaccaaggaggctgcccttac ttccaccatatccctccgggcccagggcgctgagatttctgaaatgaataggtactgcaccaccctcccctctcttcagggccagcccg cctcccaactaccctcccagctagggagccctggggatgggggtggggatgggcactgcccttagctctgctgtcattctggggaaa gccctgggacatcctgggaattggctgaagtgagcttcaggtggagggcgggctctccctaagtaagcctgaagctcccagtttgtaa tttctgctggaaatccagtcatgcattcatctccattcaccctattcattcattccttcactcattcattcattagactgttaattcactcattcatt cattagactgttagccccttgagggcagggcaatgtgaccaaaagaaacagtgggtcagagctgcatagtgagcgcccctgtacagg gggccggacgtttctcagtgcttgggtgagatgcaggcccaatggggtcagagcttctgcttttctcagaaaagccagaaatccaggct tttgcatggaatctcccagttggctcagacatttcagcccatggtgcttggggcctgagcagggctggggctgggctggcctcaggca gggaggctgtgaccttcacatttaggctgctcagcctgtgccacagcattacacctgcagcaaaccttggccctttgggagcacgatgt gtgctcagagcaggacagcaggaggcaggaggagagggaagtcagctgctctgtcctaagcatcccatggagggtgggctctgaa tggcacccggtctgcctggcctcatgaagcatcagcatcccctatttttcagagaaggacgtggagctttttggggaggtgatgggtctg aagtctcacagtggggagacagtggtttaggattcaagccaagttcttttggacactggactggccttgggccagtggtacatgtaggta ctttccaataaccacaccacacacatacagatgacagtgacactcagggacagagactcacctagtgtcttgggcctcgagggcctca ttgagctaggggaagttgagggcgggctggtgtctctgtccatcgcagtcccagccgcagggtactgacctggctcatgaagggcgt agagaaggtgatcccgcagcctgttcacagcatcacggaggacccggctcaggtactgctggggctggaggctgaaggggccgggttggagaggggctgtggggaggctgcccttgggctccctgcccatgtctgtctgtctgtccctggcagatcctggggcatggcagca ctggggacacaggtaagaacaaggggcaggatggcttttcagcagcagggctgaggggctgaggggctgctgggatgggccctg ccccttgggagcctctgcccaggctggatcttctcacctgactgaccctgctttgaacctggggttgagctgagctctcggtgcccactc caggctgcccagggggctggggcaggtctcccgctgctgactgcctttctgtattttctgaccctgcagggtgcacagatgaacccaat gaggcccttgaggcccaagacactaggtgagtccctatccctgagtgtcactgttgtctgtatgtgtgggtgggaggaacaggtccttg gtttcaggagcagggtctgttaccacctcagggtgtgaccttggacatgcctcccgtctcttctgggcctgtttccagccatagaatggg gacagtggaccgtgtgctgctcatggtttgtggtgggatgtccctgggtgttcccagggatgactgagaggtccctgggtggggccac cctctgagaatagggaggggcagggatgagctcctcacaggtgggcttcatgtaggtaggctccgtgtggagcctggaagcagggc ttggatcactgtgggggacatcagggtgcctgggctggcctcagtgtctgagttctcagagctgactcagggaggggtggctgcccc atcagtaagggtggcagactggcagggggactgtggcgaagaggggagccagggagctccgagaaggggccctgcttggtttca actgcctcttccttgctggcgtttaggccctgtgtggccagatcctccaggttttccagagaagctagaaatctagctttttatgtaaaaccc tcaatttttagtcaagaagttttcaggacactgtagacttccaaagtaagcttgaagctcccagtttgtagtttctgctggaaatccagagac atgcattcatttccattcaccccattcattcgttccttcactcattcattcattagactgctggctccttgagggcaggggcttttgtcctggtc actactgaacccttggtgcccagcacagcacctgccacagagtgaggactcagcacacatgggacaaatgaatgcatgaatgaacac acgtatatataatcacacacttcattcactcattcattcattcattcattcattcattcattcattccttgctgcagctctctctctgtgtcttttggt cctctacttccttctgagacatatgcagctgattcctgggaccactaaagcaatttaggatgagggcacctagaggactcaccagccttc ttttgggccatacttgggggaaccggttgtcctgggaacatggtgcttctctgaagagagtggagcttcccagaggccagatggatttg ggatccaatgcttgatcttaggaccagccacataatttgtggggcccagggcaaaatgcaaatgcaggccccttaaaagtttgtttaaaa tgttttcagactgtgacagcagagtatgaaaccaaaaccaggtccttctaagtctaaggctttgtgtgatggcacaggtcacactcctgta cagccagccagccctgtctgttccagcacagggagcccccagggctgcattcaggtcctcaagcggctcttgcctctcccctgctgcc ccaggaggccatggctgtggttccagagcctggtccagccctggctcagaaatggaggctgacaggatttccccacccgcttggggc cctttgagagtttccaggacaaggcccccactgtgcagaacaggattggagggcacggaatggctggttgaggcctggccccgagg tgggctgttagtgcgctggccggttccctcagggcactctgctgatcactggcccctctgcaggcctgggctgcggctgcttctgtggc tggagcagaatctggaaagagtgcttcctcagccccccaaatcctctgaggtgagccaaggaagggagaggcagggagttgagtat atgggggggcttctggggccccaggagggactgcctcctccagcaggctgtccccacccaccccagaactcatgagggggcgatg ccccactttggagagtcctctatggggaggagtgactgtgagagggggccgggattcagaaggagagacaccccccacccccatcc ccggctcacccgccttgggccaggacacagctggtagtggcggttggggaagcgcacagagcttgctatggcccatgaccttgcgc cctgtttgtcacccaggtctggagagatgagcctgcagttgctacaggtgctgcctcagacccaggtacagggaggggagctcggga gggggctgcctgggatggtcacgggctgggctcagggatcaactgggccctggaggatcccccagaggcccttagcccccaccca gactaggacggggttcagggcttgcctggagcaggctgttggcggggacaccagaatcccctccccagtccaggctgggaagcca gtgtcctggaacagacttgtgggtcttgggtaagtctcttcccctccctgggtctcagttcccttgtctgtctggtctgtttgaggattggga tgggtgaggtggtcccaaagggctccccattcagtcctgttagcgactcgaatctggccccgtgcccttgaggggttggaggaagccc tcaagggatggcgtctctagctcctgctttccccgctgcttagcgcctccaggacgcccccaggaaatggggcccaagctgcaggcc cgggagaccccctccctgcccacacccatccccctgcagcccaaggaggaacccaaggaggcaccagctccagagccccagccc ggctcccaggcccagacctcctccctgccaccaaccagggaccctgccaggtgagccaccctcaatgcagcgcctcccaccaggg agcaggatgagcaccttgtccccgcctaacacccaacctcctggaaacctggaagaaaattcaactggatgtgtgcagcctccttccc aaggcagccacatcagcttcgtgccctttcccacattttctatgcacgtccaagctttcaagctttgggtacttttcttttcttcaacacacac ctgacaggagcaaactatacatactcgtctgcactttgcttttttgtttgtttgtttgtttgtttttttgagaccgagtctcgctctgtctcctagg ctggaatgcagtggtgcaatctctgctcactgcaacctctgcctcctgggttcaagtgattctcctgcctcagcctcctgagtagctggga ttacaggtgcccggctaacttttgtatttttagtagagacggggtttcgccatgttggccacgctagtctcaaactcctgacctcaagcaat ctgaccgccttggcctctcaaagtgccaggattataggcatgagccacccggcctggcctgcttcctcttttttttttttcttaacctgaaaa atgtgtcttagaagtgcttttattcacacaggtcttactgcattctttcttgaacagctgcattaaatggatgcaccagtttttatttaaccactc agtggacatttacatttgttttcactcttttgccacagtcaataacccgtgcatgcatcattgggttcacgcacaagtaaatctggaggataa ataagtggaaatggagttcctgggccaaaggatacatatatttaaaatgggatacaggctgggtgcagtggctcatgcttgtaatcccg gtactttgggaagccatggcagcaggagttcgagaccagcctgggcaatatagaaagactccatctctacaaaaaattaaaaaaaaaa aaaaaaaaaagctgggcatgatggcatgtgcctgtagtcccaactactcagggggctgagacaggagtgagatatgatcacatcagc ctgggccaccaagtcaagaaagaaagatgggagaaaaaagaagggaaggagggaaggaaatcgggacaggtattgccacattcc atgtgtgagatgcccaaagtgcactcccttccgagaacgggtgggagggtggggggaggccgagctacctgctagggcctgaaca atagccagagcaccgagggagggtggagaggggggagaatggtcttcagggctggagacctgggctctggctcctggcaggctct gccttttggcaggcttgttttctgccttgggaacgagctgactccagcctcagtttacccatgacccttcccacctggacaccgcacactctctcccacctgcaggctggtggcatgggtcctgcacaggctggagatggccttgccgcagccagtgctacatgggaaaatagggga acaggtcagtgctgggctcatggagtggtggccctggtggtggtgggaggggtcttgggaacttgggtaaggaggaggatggttgg ccctgtggcagaagcccagctcccccagccttcctgtctgcatggcccacgttcattccacccactccctgagacatgctggcccagc cccacctgccaggagttcacagtcagcgggggaggtgcttccgtgggggagcagagaggatatggggcacagaagtggagcctc ctgggagcatcaagaagggcttcccagtggaggtggcctccaagcatgtgcaaaggtccagaaatgagagatacctagattctggga agaagtaaggctcacaggagccggcagccctatgtgcattcctgatagcctagggctccaacatgagtggccagggggagcgctca tgagctctgtcaccccttcttttccttcctcaagaaccgggaacctggcccttttttcttgggtgcaattggacgtctttatagaagcacctg gaactgggaggtgggaggcctaatctctctcctggctgtgtggcctcagctctggccctctcagaactgcagttgtcccagctgaccaa tggtcagttgtttacagaactctttacctgcattctcatacccagggctgtgagaatggaggtaaagagttccgtaaacaaagctgtctag gccccagagagaggaaggcctggccaggtcctgggcctttgtgtcactcaagagctcggcctggctcctctgagcaggtgccgctg ctgtggctgtctggggtcacccccaccttgcttcacttgagccaaggctcgagctctcaaagccccctccccccacctcaaatccttcat tggccctgggcacagcagggactctcttctccatcccatgggacaactaaggcccggagaggtgggtcaaggtcacgtgctagaag gtgtggctgaaacggggaggctggtgaagaggccgtggcaggattttcccttttttctctctccatgcaggagcctgactcccctggga tatgtgatgtgcagaccagtaagtgacacattactgagtccccatgtacatattaggaagggttgcttggtgtccttgggtctgactgtatg tgggcaggggggctgtccctggccagacctggactcacacaggggggcagttagggtctcctgggctggagctgagagagcccca tgctctgcaggccctacctctgttcaggtagccaggtagccagagcctggcccagagccagatggtggttggaggcccccggagcc cagtgtccagcttctatccagggaaaacagtcccagggctaagtcagatgagctctccttcctgctggtaatgcccagagagcttcccc catcccctggcaaatgggtttggccaagctgaagggagaggggtggggtctcactgggagaaccaggctccacagcctccagctcc cttgagctggagggggtccctgtcacctgcagctgggttggggaggcatatgatgccctttatcccagccctgagccaggaatcggg gacagaggactgagaccaggaaggggctggaggcaggatctgttctttcttcacttcatggcctctagagaggcatgcggtggacag gcattcattcattcattcattcaatacatagttaaggacagcctactatgtgccaggcctgtgatgggttctagttgtgagcagagttcagg ggctgtgggtggtggcatcccctgttccatcctgaacacaagcccttcaaccacaggcctcaggctgtcccagggctctgagctctgc cctgtgtctgcaaggctgtgtccagccccagccccgctagtctgatggtggtctccatgagtgccctgaagccgggggtggccgccat ggcctggagcacacacagcccagtggactgtggagctcaggtctgatggagaggcgtggggggcgttagggttgatggtaatcatg gcgatgacatgaggaggaggagagctggcctgtctcagagcagggcataagcagtctctcatggaagctcgtggccaacctgggg cagcagagcagagaccctcatggctgttggggcaggcagtgggcagagctcagagaaccaaaagcacctgcccagggtcacaga gcaaatcgagtccacctggcctggcagccacagcttaaaggaactaaaaacgtttccttggggcagagagccttcagagggtctgcat ctgaggatggagcatcagaaccctctggaaggaggcaggattccccgccagctgtggccagggcctccctgcaatgcctggagcat atgggttatgggtggacgctgggtggccatgggggcaggctcatggggacagggagaggagctggcttggtttcattttctaggactg ccgtaacagatgtgcgcacactgggcggcttagaacaacagacactgattctcccacagttcaggagccagaagtccagggtcaag gctttagcagggttggctcctcctggggctctgagcgagacatgcccatgctgctctccttgctcctggtggccacttgcggtccttggc aaaccttggcttctggctgtcctccagtgcccgcctccgtctctacacagcctttacctctgtgtctgtgtgtgtcaagcctccctctccgtt ctttaaaaaggacatcagtcactggatttagggcccaccctcaatccgggatgatctcactttgagatccttaacttaatcacacctgcaa agaagctatttccaaataaggtcacattcacaagtaatggggttaggacttgcaattatctttttgggagacaccattccactcacgtccca tgtcatggggagactctgagaggctgggggatatgtgtcccttaccttccccattcattcactcatccactcactcactcactcactcattc cctccgcctgtccacccagccacccaccctgtcttctcctttgtttcaggggtgatgggagctggaggtctctgaaataagggagaagg gaatctgggagagctcagatggacacatggatggaaggaaagaaggatgccctgaagagaagaccttcccgggggaggtggcca ctgacaccccaccctatttgaacagcgaacccctccctctccacacactcccaggcaggacaaggggagccagagtcacctgcacc agccccagagcctcccagaccagaacaaggggaaggccagccatgggcagcctgcaagatttaggctagacagcaggacttaccc ccagagggctgtggaaaaggcccatagccgggcgtggtggctcacgcctgtaatcccagcactttgggaggctgaggcaggagga ttgcttgagccccgaagttcgagaccaccgtgggcaacacagtgagaccctgtctctaaaaaaccttttttaaattaataaattaaaaagc cccatggatggaggactcagtattgagcatctctttgagaggacgcgtgcacagcccccagtgtggtacctggcaccgtcagcacctc gacaggatacagtttttcccagaagaggttctccctaggcctggccacactctctccttccaaggcttggggacaccaatgagcagcaa caaatgttttcattaacattaaaagagtgtaaatgagcacatcaaacctgtgattgcattgatattattgcttaggataaagctaaaagaagg gtttaggttttaaaagtgggtggactgaatagccctgggcttgcggtgttgggttgcaggaggagaggaattagccaggggtcctgag ctgggggtcctggctcccagctggcccacatgtgtctttgtttgggccatggagtgattttaaaaattagaattgattgccaacatttaaaa atcaggtagctttacatggaaaaatctggcttctcttgaaaactgggtggacttggctgtcatgggcccacactgtgcttgtgaggacagt ctgggcagctgctggcacctgaaggcaggtggcacctgcctcttcatttgtttacatcatcaccctggctgggcaggcctttggatttga gatccctgaggtctcaaactggtgatgaggtggggcaatcccagtagagggaataaagtgtggcatacaggagagtggtgggacttc agctaatctggagcccagcatccttaatgcagaagataaaactagagaagttggggcctgactttaggaaactaagctgtaggcaatggggagccattgaaggtgattgagcaggggagtgacttcagaaagattactctgaggctgagcaagcattggagacagggaaagcaa tcaaagcagggacacttccaaagcacctggggccaactatgaaaaatgtcctaccccagactttctgattcaagagtaccgggagctg ggaatttggttctgtgttgttttttttttctttctttctttcttttttctttgatgaccaagcaatggttatggaagaatctgctttattgagaagcacct gctttctcagatgattctgatgacggggtagaggctgggaccatactttgagaaacatccagtgagaggttagtgctttggggccaggt aggaggagatgtgagcctgagccagggagaggcagaggaaggaggtatcatgacccaggcctgccagaaaggatggaggtttgg agttagggctcaggctttgacagagacccaggggtccaggtggggaccttagcctagaattcatgtgccctggctcctgcctgctgatc ctggaagcaggggctccttcctacatgaggctgttgactttctttttttttttttttttttaagattgagccttgctctgtcaccaggctggagtg cagtggtgtgatctcggctcactgcaacctctacctcccgggttcaagcaattctcctgcctcagcctccctagtagctaggactacaag tgtgcaccaccacgcccagctaatttttgtatttttagtagagaatgggtttcaccatgttggccaggatggtctcgatctcttgacctcatg atctgccgacctcggcctcccaaagtgccgggattacaggtgtgagctactgtgtccgaccgaggctgttgactttcactggccagggt cagaacctcatttgatggccctgagggcagagaggccactgaccaggaggtggtgtgccagggtggagtggcagtttctatgggaa ctagggcactcatagctcctgagcacctgggtagggccacggagaagctgggcgagctatgaatcagctgcctgtagagaggggca gttggagctccaggggtgggccaggggctcctgcaccacgcctcctgggggcctgccctgggggctgtgtggcagcatacccaatc cctccccttccaggttggtgatcttgggcaccgtcctcctctcagtgctgcagaatctttagctgtaaaatgggaacagtataggacacat cttgcaggcctgctgcggagatggaaagaggaaatgaggccgggcatggtggctcatgcctgtcatcccagcactttgggaggtcaa ggtgggtggattgcctgaggtcaggagttcgagaccagcctggccaatacggtgaaaccccatctctactaaaaatacaaacattaac ccggtgtggtggtgcatgcctgtaatcccagctactcgggaagctgaggcacgagaattgcttgaatgggaggcggaggttgcagtg agccaagatcataccattgcactcaagcctgggtgtcacagtgagactctgtctcaaaaaaaaaaaaaaaaaaaaaaaaaaaggaaat gcttgtaaaacacccacgcactaaataggacctaaaaagaaaatgttaatggaaaggaaaggaaaggaagtgtggaagcaaggaca gcatcaattcttaaggacggcaccccataaggagtggccactgatgccagtgaggcgctcgtggggcacatttatcacctgggatagt gcactgggaaaacgggagtcaggggacctggctctgcctccaccttcctgtgtggcctgggacaagacactcagcctctctgagctg gctctctttaaaagggggaaagtactgttcctgccctttggggctgctgtgaaagctcacgtgctgtcaggcaagatgttctgttccctgg atgtcccctgctatgcccaggcaggaactggcatcccaaggggctgcttagagtggagacatggcactattgctggacagcagcata gactgacacagacaaagcgtcccaccaccggattcagggtcacacttggaggagcctccctggccaaaccccaggggcgtttgcag ggccagaggagccgggagcaaccatggaaacagctagttcaggatgctcttggggcttccctagagggcaggcagcacctcatctg tgaaacagggataatatctgtacctagaagtggggagctctgtgcctggcaatggaagcaccccataaaagaaggtatttatctgtgtct cagtttgctcatctgtaaaatggggagaaaaacataaaagagggaaaataaaatggggatgcaaattataaccccaaaatgttgatgtg aggactaatcgagttattccacgtaagccattcagcacagggcctggaacatagtaagtgctcaaaaatgcagctgctgttactattattg ttgttatgcaacatcatgaacaataacagccaaagtctcaggataagacttgaggccggcttccatttaaagtcattgttactactacccag gtatggtggtgcatgcctgtagtcccagttactcagaaggctgagtgaggtgggaggattgcttaagcccaggaggtcgaggctgag gtgagccataattgcactaatgcgctccagcctgagcaatagagcaagaccctgtctcaacaacaaaaaagtcactattactaattcatg tttggctgaggtccaccaagatccctggagctttcgggggacacctagtattcaggcattgtcctttgtccctgacacctgcctgggggt aggcttaggttcctgatgagactgctctgcactccctttgagcatctgccaggcttataggcccagggtgtttttgttgtcaaggaccagg atacttgaagaagtgttgaaaattagactcccatctgaaattgggaccacccccatgagaaagatgagtttccctggagggcggagcct ctagacccaggagggagggaaggactctcagaagcaaatgccgaacgtatgggcgtgctgtcacccgggggtggcatctacagag caggaagcgggcaagctctctctgcatcccccaggacaatccagtccacctgcacctggatgtggccaggtggaccctgaacctgg gccttttggcatccctgtgatgctgtgattggaagcattgtttctgcctctgcacaaacctttaagaaggaatcccccgtgagaaacaaac aggatcaaccataatcaaagctctgtctcaagaggagaaggggggaaaataatggagacagagtgaatgagcacaggcttcaacct atcaccggaacaccgatctggtcccgctgaaaatggaaagtgccatttccccactgcatgactaagcactgaggcactgctggcgaca gccagggctccctgcaggctccctgggactcagaggcccaagcccactctcccgcacatcccataaatggctcttcctgtgttccctg cagtcagcatccttcctggaggacaagtggagcctgaccttgtcctagaggaggttgaaccgccctgggaggatgcccaccaggatg tcagtaccagcccacagggtacagaggtggttccagcttatgaagaagagaacaaagctgtggagaagatgcccaggtggggagtc tgagaagaccctaaaagattagttattgtttgagaaagtgccaagcatgcagtaccttatctcaaagatgaaaaggcacagagcaggaa ccaagcctgcttccccctgccttcatacagccatccttctcccatgggtagaatggctaccaggttcttatgtgccactccagagacagtc tgggtacatgcaagctcgcatctccctccttttttctttttgtacagaagatggcatagcacatgtacctatctacaccttcctgtatttattcaa caatataccttgaggtctttccacattcaggatatagagggcatcttcatttttcactgctgcatagtattccatcatatggctgtaccataatt tattaaacctcttcactattgatggacatttaggttggttccaaaattttgttattatgaacaattctgcatcaggcactcttgtatgcatgtcttt gcacatgtgtgaatacgtctgcaggagatttctagaggggaattgctgggtcattggtcatgggaatgtggaattgtggtcatgattgcc aagtcgccttccacagggatgtcgcaatctttgttcctactagagatgcttggtgacagtgggaggtctcgggttctgaagcccataactt acatagctaagatggggaaactgaggcccagagaggatcagaggctgacctagggtcaccaccaggacagtggctgggacagtcccctcatgcagaggagagggtttgttgccagagagagttcagggttgctcgggtggggctccccaggttgagccttggttggaaagtcc aggtctgggctcaggtgtcatgggacatcctacagaaaatagaggcaggaggtagactcagcggggcagcttcgggtgggctcacc tcctgcctctaaccccaactcagagagctgtcccggattgaagaggagaaagaagatgaggaggaggaagaggaagaggaggag gaggaggaagaggaggaggtgactgagtaagtcatttgctcacctgtctgggaacggccctggggctggggttggctgagctcctg gggtctgggcactctttgctgctcatgggagctcctttctgttgttctgtgctgtgtaaggctgggctttgaccaaagaggctgcgagaag ctggggttcccctgctgtgctcatcatgacctgggaatcatattacccaccactccacatccccccccacacacactagaatgcagcctc caagagggctggggcttttatctgttgctttcactgccatgtccctagagcctagaacagtgcctggtacacagtgggcaccaaataaat gttagttgtctgcatggacatatgcagtgggctcccagttctccctttattcacaaagccaaggctgcctcccctccatatccacccacctt ggttgcctttgatgccttctagggctgtcctggattctgctctactgcccagtaatggtcgatcgtccacccacatccatatatttcaatgtta aacacacagaagagttcagttaaaataaaaattaaacactggagatcaggaaacactatggagggtgagtggaaggatgactggaca gggggatatgcatggggaagagcaggcagttggacacctgtgcctggagttaggagcaggagaggtggggctggagatgtgacat tgggattctgggcatgtaggtggcattgaaagctgtggggctcgaggagatgcccagggagcgagtgtggatagaggagaggctca tggacaagaccagaggttggggagatgaggaaccaaaaaggactgaggaggaggcaccagtgagcagggcagagacaagaag agtatggggcccagaagccaagttagtgtttcaggaaggtcgaccaggcgtggtggctcacgcctgtaatcccagcactctgggagg ccgaggtgggtggatcacttgaacccaggagttcgagaccagcctagccaacatagcaaaaccccatctctacttaaaaaaaaaaaa aaaaagaagaagaagaagaaagaaggactgcctcaaaagcaggttgtaaagccttgaacttgttttcccaaaggcgatggggagcc attgaaggttcatgagcaggagagtggcatgatctggcagctgtgttggagaaagattgagtcagggagaacagtgaggagactgtta tggtgggccaggtgacagtaagttactctctccttgctccagtcccctggcattatgtccctgtgtctctgaccaccatggtacctcatgac ccgcgtcctttctccctgacctgtccctctggccatggcctgagacaggatagcactgtgagtgttaggagcacaggctttggcatcag aaacgccaactccactttgcttctgttagtgaaatgactggggcaagtggtgtacccgctcagagcttctgcctccacatctgtaaaatgg gggataataaaaggctcctgcagaggggtcatcagggccagcggtgatgatggaagttgtaggggagtaagtgccccatcgatggc cactgatgttctcgtcccccagtggcctcttttcgttccagtcaggaaatagttaaacaggatttcagagctggaagcctgttcagtgtcag ctgcttttagtccatccttactcagttcacatagactctttccatccttgacaggaggccacacagcctctacttgcttacctccaatgacgg ggagctcacaacctctcaaaggctgtccatgtcatccatatgccccgctgacttgtgaaattatttcttagaccaaaactaagcctgcttcc ctatggcttctgtcccttcctgacttgtcctaattctgccttctgagccaccctgcagtcccagcaagatgaggaacttagctgctgcctgg tttcattagagcagcttgtttgccctctgctttctatggagctgtgcttctgtgtggggtgtatagctacaagtggctgccttgctgaacttgt acacctaacacccaggctggaaaacccctgtatttgccactgctctggatctgagacaccctttgaaggccagtgatcaactatgctcac atacatgcatgtgcagatgtgtactaactatacacatgttctcagaatgcagtacacatgggcacacacacacatatctgcactataccca tatgcaccagcacaccaacacagacatacatgctgggttatgcacacacttacatgtacaggcatgcacacacgtatacccacacacat gcttcatatatgcacaagcacaggcagcacattcatacatattcatacatgtgtgtgcacacaggcacacatgtacatacttgcacacatc cacgtgacagccttgcacaaatgcacatgcatccagagttaccggcagatgctgcaggcgtgaacatcttcatacaaatgtttacccag agttttgaacttggggaacttttgggctccaggcccagggagggacaagggagtgacagagttggtgatcacagggccagggccag agggtgtgaggcccagggcaagaaaaactgggctgtcaatggctggggaaggatgctcttggccagctttgcagagacacaggtta ctgatgatgtgagggcctggtaacgggcagatggtcgcctagagacccggtatctacaggcaacagctcagctttccctagcaaccg aggccttttggttccacctggcctccctggggatgcttctgccattccctgcacatctcctcaatctgattagaagcaggaaggattgctg gctcctctgccactggagccaccctggctcgggagctgggctcccacttgcatgcaccgacccaggcttgggatgaacgccacagc ccccagctgaacccctgcaccatgggcatcctgactggtcccgaccgagagatccaagtaggtggctgcctctccctaaggctcccct ccttcctgcttttggggctgggatttcaggtggacagtctgctgaaggtgacagtgaggaccttaatgttggggcacggcctgcgggat ggcacaaggcgggaggagtggacgggtaagcttttgagagctggctgccactgctgctccctggatcttaaacttggaaagccggag ggtgtctccaggaagccaacctcagccaatctgtctgttcctttgcttggaaatccctgggacttgggaactttctggaagttgatggcttg ctgcttctgggctgcacaagagggggccttggcaggcactgggcaccctgtcaagcaggggccatacggctttggtaacatggccc ctagacagcccccggtgatgctgacagcgccaccactgggctgtttaggtgttttgatagctggggatttttgtgcaaaaggtttctcaat ccacgtgggtgatttcataaccctgcagatcaccgcctcctctgggattaataataataatgacaagaacaataataatggcctccatttg acttacgatttttttccatgccactcacggctaaggcagcctgaatggcagattctattattctctcatcccatttcacagatgagagaactg aggttcagagacagagccctgccagtaagcgtagagctgggattcttggcattacagacaaggaaagctgctcccaatgtttagtgat gaagaatccagaagaatcaccagaacaacatctggcagtctttttttgacttttctgtgagattctcattggcccttctgatcttactcaaata ggtcaggctgcacaggccaggcaaatcctgcccatttgacagatgagcagacagaggcagcccaagggcttcgaggtgacttgccc aaggccatacaggggcttctggtctgacactggaataagcaggaccccaggaagcctctggctgggcctgtgggcagttcattttctg agccttgtgctgtaggcagagagccgagacagaacctgtatctatttcaggaaagtcatccagtcatcccccttgcagccggagacct gtctttaacccatttgtgcaaggctgacttctccttagtgtagaaggcacagtctccaaaatactcagaggcaggcagtgaaacagagcattaagcacgtggccttggggtcagtgatgcagtatcttatccaggcccaccttgcacaggcggtatgaccgtggattggccatcgaaa ctctctgagcctctgttttctcagcttgcatgtgagcagtaacagttacgatctgtcagggttatgtgagatttgagtttttttttccctaggctc ttttaaaaagttaattattttttcttttcttgttttttttttttttttttttttaaagacagagtctcgctctgttgcccagggagtggagtggcatgata ctggctcactgcaatctctgcctcctgggttcaagcgattctcctgcctcagcctcctgagtagctgggattacaggcatgtaccactgtg cctggctaatttttgtatttttagtagaggcgggttttgccaaattggccaggctggtctcaaacttctgacctcaagtgatccgcccacctc gacctcccgaagtgctgggattacaggcatgagccaccgtgcccagcctaattattttttcaattgacataaattgtatatatttttgtgtata attgttgttttgaattatgtatacattgtggaatggctaaattgagctaattaacatacacatttcctcatatgttcatcattttttgtggtgagaa cagtttagatctactcttaacaatttttacaaatacattgctattaactagagtcacaggctgtataatagatctctcacatttgtgcttcctatct agctgaatctttgtaccctttaaccaacatctatctccccacctctcacccctccccagcccctggtaactaccattctattctctccttctgt gagttcaacttttttacactccacgtataagtgagatcatatggtatgtgtctttctgtgtttagcttatttcacttaacataatgtcctcaaggtt cacaaatgacaaaatttccttcatcttaaaggctgaattctattgtgtatatataggacatttccttgatccactcatctgttgatagacacttg ggaggctgaggcaggtggatcacctgaggtcaggagttcaagaccagcctggccaacatggtgaaaccccgactctactaaaaata caaaaattagccgggtgtggtaacatgggcctgtactcccagctactcgggaggctgaggcaggagactcacttgaactcgggaggc tgaggttgcagtgagctgagatcgcgtcattgcactccagcctgggcaacacagcgagactctgtcttgaaaaaaaaaaaattaatgct gcaatgatgggagtgcagacacctcttcaacgtactgttttcatttcccttggatctgtacccaacagctgaatcatatggtagttccattttt aacttttggtggaacttccgtattcttttccataatggctgtgtgaatttacaatcccaccaacagtgtgtaagtgttccctttccactgcatcc tcaccaacacttgctatcttgtcttttttcgggtgggggcagggcgacagacagctggagtgcaatggtgcaatctcagttcactgcaac ctccgcctcctgggttcaagcaattctcctgcctcagcctgccaagtagctgggattacagaagtgtgctgccatgcccgcctaatttttg tatttttagtagagacagggtttcacaatgttggccaggctggtctcgaactcctgacctcatgatccacccacctcggcttcccaaagtg cttggattacaggtgtgagccactgcacccggcctatcttgtttttttgataattgccattctaacaggtgagaagttaatatttcattgcggtt ttgatttgcatttctctgatgattagtgatgttaagcatcttttcatatacctgttggccatttgtatgtcatcttttgagacaccgaggaatgtct attcagattctttgcccattttttaaaatcagtttttttccttacaattgagttgagtttcttatatattttggatattaaccccttatttgacttacggt ttgcaagtattttctcccgttctgtagggtgtttcttcactatgttgattgtttccttggctgtgcagaagctttttagtttgatgtaatcccattttg tatatttttgctttcattgcctgtgctcttgggttcatatttttaaaaatcattgcttagaccaatatcatagggtttttcccctatattttatatattg gttttatagtttcagattttacatttaagtagtgttcaatcaatgtgggtagggtttttttttttttttgagacagggtcttgctctgttgcccaggct ggagtacagtggtgcaatcacggctcactgcagcctccacctcccaggctcaagcaatcctcccacctcagccttccaagtagcttgg accacaggcacatgccaccacacccagctagtttttgtatcttatagagttgggattttgccgtgttgttcaggctggtctcgaactcctgc actcaagcaatcctcccatctcagcctctcaaaatgccgagattacaggcatgagccaccacatccagccatcttcttgagtttctttcgtc gatgttttatagttttcagcatatggttctgtcaccttcttgattaaatttattcctaagcattttattttattttatagctattgtaaattgggttgtttc atttctttttcagatagttcattgttaatatattgaaatgctactgaattttctatgttgattttatatcctgcaactttactgaattggcttatcagttc taacagttttttggtggaatctttagggtttttttttatatataagatcaaattatctgcaaagaggaacaatttaatttcttcccttccaatttgga tgcctttcatttcttcctcttgcctaattgctctagccaggacctcttatagtacattgaatagaagtggtgagagtgggccaggcgtggtg ggtcacacctataatcctagcactttgggagaccaaggtgggaggatcacttgagcccaggagtttgagaccagcctgggcaatgtag caagaccctatctctactaaaaataccaaaccaaaccaaaccaaaccaaacaaaaacaaaaatagccaggagtggtggagcatgcct gtagtcccagctactcgggaggctgaagcaggaggatggcttgagcctgggaggttgaggctgtagtgagccatgatcatgccattgt actccagccttggtgacagagtaagaccctgtctcaaaaataaatgaataaataaataaataaataaaaaggaaaagaaaaataaataa ctagtgagggtgggtatgcttgtcttgttcttgatcttaaaggaaaagctttcagcttttcaccattaggtatgatgtcagctgcgggcttgtc acatatgacctttattgtattgcggtccatttcttctatatgtaattggttgaaagatttgatcatggaatgatgttgaatgttgtcaattacttttt ctgcatctattgagatgatcatatgattttgttgctgtggtgtaccgcgtttatagatttctcaaggtaatttggaggaggatctggggagact tttcatgcagactgcctggcacgcagcccaggagccgctgtggcaatgtgattaccgactgatttctgctgccaagcggctggctcca gaccgactaggcaacaatcccgtctccaccttcctgccccaccgtccttgctcaggagggctgagacttggccctctgggtagagaat gatgcattcaagtttcctgggtggcagacaccccagagcatcctctgtgaacccctggatgacaggagtctgaggggtggggagggg gtgttaggaagcaggaactaatcctaactacaaaaatcacattagtccctggtcctgagtcatcattttaacccagaggtcgcacacgga catctaatttagcctggagatgggctcttttggctactacatattaataaagatatattaatttgaattatcagcatctaaatatcagacaacttt accttagaaacatatatttcaggctctgtttgaaaatggaatgatctggccacctgggcccccttttccagcggagtggggttgctgcctt caaatggacatgggttcgctaatttgccacagccctccctgcccactgcaccctcacccctggccctatcacttatgtatgcctgctgctt gggcccggcacccatgaaaacttcagcagttgcagtggctctgatctcatggcatgggaagaaccagggcctgggtttgaggtttgtg tggtttgggacaatttgctgtgtgattttgccaggtcacttcacctctctgggcctcacttttgtcatgctctacccatcaccaagattgcctg gagactgggagaccccagatgacagctcctccaaccgtcctccatgcatttctcgcacatgttgttctcatttcctggacaaccttttcctc cccaccattctttgtggtaaaatcatcatcctggaggctcagcttacaggccacctcctctaggaagccctccttcctagcagagttgtggcatgtccttcacagtgccttgctgagtgcatgtctcagatggggtgattgttagtaggacactgccctggaggctcctgaggttgagact ggttcttgggcatctctgtaccctccgctcctggcacagagcgtggccctcagcagggctggggaaacctttaacaccctaacccagt ccttttatcctccccaccccgccccgggtcacagggtgctgctggatagctgtgtggtgtcgcaggtgggcgtgggccagagtgaag aagacgggacccggccccagagcacttcagatcaggtatgagaagcaccaggcactggcacctctgaggcctggcttggcattggt ggaaggatggggaccttgtccactttgcttacagggggagttttggctggtaagatgcccgcaggacagagggagcagggtggcgg gtcctgcgaggccccctcttggcaggggtcagatagcggcccaggccgtgggtggggagacgggtggacccagctcgagtcctcc tgtgggtgcctggcttggggcaagtcgcctccccactctgggtcccagtggccacctgtgcacgagggggctacagacgcctccctt gggcctgtaggaggacccaaaagggcttgcacactgtgaagagtggtgctgtcgatgctttgttcccccatgtgccaggctgcgagtg tccctcatcacaggtgacacgtccagctgttaggacccgtaggtcctgggtgcactgccctctacaggttgtaggctactcccaggatg cccagaaggtgggctggcaggacctgactggctgggcagggcaagaagcagaactgggcccatgtggacttctgaaggcagatct aacaggagctgctgagactagggctgggggctgatggtcacagtttctgttttcactttattctctgcagaagctgtgggaggaagttgg ggaggaggccaagaaggaggctgaagagaaggccaaggaggaggccgaggaggtggctgaagaggaggctgaaaaggagcc ccaggactgggcggagaccaaggaggagcctgaggctgaggccgaggctgccagttcaggaggtaggtgtggtggtggtggtga tgggttgcgcaagaatgtctgaggtccttttaggctggaggcagccttcgtgcaggggcaacccctcctcctctccctccatctccttcc ccccattgccccctcagccaccctgtcttcaggaaaatcccatccttggtgtcttggggccacattatcagagcctactgatacatctcaa aggttctaaggaatttcactgaggcccaggactcccagtggcgccggggaaatcccgatcccagccaactccttccttccagcctctc cctgccagcccctcctgcaggcccctcctaaccagcccctcccagccagtccctcctgccagcctctcctgccacttccgctcaatga ggatgcagctcttgggctaccgcccagtgagggtttggctcagacatcacatagctgagtgaaaactggaaccagaacttcctaacag agtgtcctggactccatgtctctttatgagcttttcctgatctccagctgtacccagcacatacattttgctgtgatttaccaccctgccctca atcctgagaactcaagcaatggcggagaaagagtcccaaggagtccagcaacagctggaccctggagctgggacaggtgggacc caacccctggccggcaggtgttgcgtgccttacactccacattccccacgcaatgggtcttcagccgctctgtgaggcagacagacca ttttacagatgagaaaactgaggctctgtgtgtgcggatttggccaagaggtcaaggcacagacctgcatttcactcctggctcctttctc tcttccttgctgtgtgacccagagccagttctgtgacctgtctgaaccatgacttccaaatcaagaaaaaaggggcaataatagtaccta gctcacaggttgctgggcagagtaggggagacaatgtgtccaaggcaccccaaactgtggctggcactgagggaggagtcaatatt gggacgaggatgtgtctcggccagcgagtggttgaaatgggactggacccaagtgtcctaaccctgggtatcctgagcttgctcccta cttggtctttgaaaagtgttctctgttaacttcaacacatatttataggaattcctactaagtccaggcctggggctgggtccccacatggat gaaaaagaccaggagcaaacaggtgcaggcttggttcagtggggtgggttctgccccttcccaggtgatttggaagcaggtggtctg ggacccacactctgagaaatgctgaggcatagccatttgcactcaaaagtatggacctagacccagcagctttggcgtaacccagga gctggttagaaatgagtaacttgggccgcgatccagacctgctaaatcagactcctcgtggtaacaagatctcctggtgatttgtctgca catttgtgtttgaggggcaatagccttgagccctggtcctcaaacctgacagcccattggaatcacctggggagatttaaaaattcctgct gccaggcctcatcccagccattaagtcagaatctctgggaatgggcctgggtaacagaaattgtcaaaccttcccaggtgattccaatg cacagcccagttggggaaccagggccataggaagagttactgcttctggttggaagtgaaattagttgacctccagggctgctttggac tggaaaattctaccaggctaagaatgttcgagtcctgggtttcttgggtctaagaatctgagattctgtgagttctgggttctttgattctaat atttttgggaaatgctgccggtggttggtaagtggagggagcatgggaagttgggggcctcctgggctgtccctcggtgctcctgggt gggccagccacatgacctgatgacacattagacgttgaactccagggacaagagcgatgcttctggctttgggtagcactgctgaaaa tgcaggtaggcaccaggctggtcgtgttaggctggtgggaggacccccacctattgtctcctccaactctgtgctcctaattgcagtccc cagggtcactcctctgcctgagactctctggccccagcaccatggctacaccatgtcagaacatggagctgaattgcctagtccagga ccagatacctggccaggtggcctctgcctctgccctccaaaacctggtagagcatctccctaatgtgcccagctaccgcatcccaatca cccgcatccctgtcctcacctcccggagaaccagcttgtccaactccagcttcgccaaggagaccaggagctccatccgccaactag gtagcacgatcttgcccacccagcattctcatatctgtggcgtgtgcttctgcatggctggggctgtcttcctgcctgggtgctgtggtcat agtgtggaccactagttgcttatggaaaaccatctccttttcctctgccgatcaataccttcatcctgctttgtgatagtgacttgaggggttg tagcccccttctcttcctgcctaaaattccaccacaattagcaattcatttatttttttctctctctttctcttcctctttctctcttttcttttgttttttttt tgtttgtttgttttttgagacagagtctctctctgtcacccaggctggaatgcagtgacatgatctcagttcactgcaacctctgcctcccag caattcttgtgcctcagcctcccgagtagctgggattacaggcatacgccatcacgcctgactactttttgtattttagtagagacggggtt ttgctatgttggccaggctggtctcaaactcctgatatcaaatgatctgcccacctcggcctcccaaagtgctgggattacaggcataag ccactgtgcctggccagcaattcatttctttgggttaaacatatattcctttctttaggcacattgtttttatacaggtaaaaccttataaacctt agtttaaacaagtcacttgctcagatctctaggccgagaggcttggacagggagtctgggatcggagtggaggggagtgaaaggag ggaaatggaatagaagagtaatgtgggtgctgattgcagacagaacccactgaccttgacagtttgctgctcagcaaattccatttcag gaaagaagctccaggtgcagatagtcatcctcagaggctagggtggagggagggctgtctggatccattgattgtgacaaggtcccc gtagtggcctttgggctgctactgctcactccctgatgacaaagagctcaggaacatactggcaccctggactaacccctgtcactttctgacttaacccaagccagacatgcctgcttgcccttttgattggcttgtttgtttatttgtggttgacacggtgcagagctgcctctgagatca gtccaaccaggcaggacagaacatttcacgggaagacgtccactgttgtccttctctcctctccctggcttgaggtgggcacagagag gagatgaggtggcatggaggtcatccaacagtaaggggtgccaccccggcctccatcttctggtaggcaagccttgaatctactactc caaagcagcccacccctatcattgaggatgctgaattgcttaaaaaaaaaaagaaaaaggaaacaaagcaaaaagaccccaaagcat cccccagctcccaccgccacaggccactccctgataactgctaaatctgtaccaattctatttttttttggggggggacggagtcttgctct gttgctcgggctggagtgttgtggcaccatctcggctcactgcaacatccgcctcccgagtagctgggaccacaggtgtgcaccacct ttccgggctaatttttgtatttttagtagagacagggttttgccatgttggtcaggctggtcttgaactcctgacctcaggtgatttgcccacc tcagcctcccaaagtgctgggagtacaggcatgagccactgtgcccggcctcaattctaattttatttgcttactatttttctttcagtggatt ctcttttttatttaattaagtatatttaaacaggaaaactttattactattcaaatggaaaaccagcattgcttgccataaacagaggcataaat aaaaataacctcaaatacactgaaaacacaatggtgttgctgttttctaatgagacggatgggcctgctgcaggctcgaagctgggcct gctctgttagccacaagtgggattaggctgtgtgagagaggctctaaggccacagtggcaccgacccgagacttcctctctgggtattc actaaaagagggtgtaactttttttttttttttttgagatggagtcttgctctgttgcccaggctggagtgcagtggcgcgatcttggctcact gcaaccactgctgcttttttttttttttttgagacagaatctggctttgccacccaggctgaagtgcagtggcgcaatctcagctcactgcaa gctccgcctcccgggttcacgccattcacctgtctcagcctcctgagtagctgggattataggcacatgccaccaggcctggggaatttt tgtatttttagtagagatggagtttctccatgttggccaggctggcctcaaactcctaacttcaggtcatcctcctgcctcagcctcccaaa gtgctggaattacaggcgtgagctgctgcgcccgaccaggagtaacattcttattctaggttatactttgccttgtctgtaccatctgagat catctcgtgtcctccccagtctctgggagccccacacttgggaaaatgttcccggggacacattgacacagggggctgatctagctgtg ctagagtacatgatgattctagatgctacagacaaggtaatgagaagctgtagtaagaacattacttccattttattttattttttattattttgtg agacagggtctcgctctttcgctaaggctggagtgtagtaggcgcgatcttgactcattgcaggttctacctcccaggttcaacctatcct ccaacctcagcctcccaagtagctggaacactgtgaccagccgatttttgaatttttttgtagaggtggggttttgctatgttgctcaggct gctcttgaacttaagggatccttccacctcagccttccaaagtgctgggattacaggcatgagcaactgtgcctggccccctttacattttt aacaacttctccattcctgctcatcctcttctccagaggatttagggttcactccattaaactatagagtaagtgtaagcaatttaacagagt gccggcttttcttataaaggccatttgaaaaaaaccccatatcggctgggtgcggtggctcatgcctgtaatcccagcactttgggaggc caaggccggcagatcacctgagatcaggagttcaagaccagcctggacaacatggtgaaaccccgtctccactaaaaatacaaaaat tagccaggcgtggtggcaggggcctgtaatcccagctactcaggagtctgaggcaggagaatggcttgaacctgggaggcagaggt tgcagtgagccgagatcacaccactatactccagcctaggcgacagagcgagactccatctcaaaaaaaaaaaaaaaagaaaagaa aagaaagaaaaagaaacaaaaagcacatatataattatttcaaaatctccctgcttcccgcttttggggggtgtatccctgcctgccttgct ggtgcccacacctgtgcccagctgctattgggtagacccccctaggtgtaggggagaaggaggaatactatcccagcctctcagtatc ctctgagcttggacttggggcagacctgtccctgcagtccctgaggtgggtgatggagcccaaggtagggccacgtccaggctccct ggagaggacctgattctcagaatgtgggacacagtacagggcctctcagaggtccagagtcaggctagctgatgaatttgggggccc ctggcatgtacaagaacaaggacataaccaagcagttatccaaataggaagtagcacacactacacagtctgttacccttgattggagg gaccctgcaaatgtctccatttgatgctcttcaagaatgtgaggccagccgggcccagtggctcacgcctgtaatcccagcactttggg aggccaaggtaggtggatcacctgagctcaggagttcgagaccagcctgggcaacatggtgagacctcatctctacacaaacacaaa aattagccgagcatggtggtgtgcgtctgtggtcccagctactcgggaggctgccgggaagatcacttgagtctgggaggcagaggt tgcaggaggcagaggttgcagtgagctgcaatcgtgccactgcactccagcctgggtgacagagtgagacccgcccccccttctcaa aagcaaacaaataaaataatttttaaaaagaatgtgaggacttggctgggtgcagtggctcacacctgtaatcccagcaactttgggag gccgaggcaggtggatcacgaggtcaggagatcgagaccatcctggctaacacggtgaaacccggtctctactaaaaatacaaaaa attagccgggcgtggtggcaggtgcctgtattcgcagctaattgggaggctgaggcaggagaatggcatgaaccctgggaggcgga gcttgcaatgagccgagactgtgccactgcactccagcctgggtgacagagcaagactctgtctcaaaaaaacaaaacaaaacaaaa caaaaactagccgggcgtggtggcgggagcctgtagtcccagctactcgggaggctgaggcaggagaatggcatgaacccagga gaaggagcttggggtgaactgagattgcaccactgcactccagcctgggagacagagcgagactccgactcaaaaaaaaaaaaaa aaaaaagaatctttaagaatgtgagtacttgtgccttctggctcctccccacaggcctgggaaggcatggtcaccctaacttggggtgg ggaaactgaagctcagagaggggaagcagctgactcaggctgtgctaggccaggcgccaggactctggctctcctgtgtcctctctg ggggctgccactgcctcagctcttggagcttgctcaagtgcagggaaaccaaactgggctttgagagctaggagagctgggctccag gctgtgtaggggagctcccgtggtttctgtggctttgtggcttctttcgtggctcctgggctggtggctgtgaccctcagaggaccccag gcaccacgttcctgtaccccctgcatggagcttttctgtgtgccccattgggaagtgaagctatcagagtcccctccaaacatgctttctct ggttttcttcaagaacacagctggactcctgggacccagcttttgtctctctctgactttctcctctccttcctctcccccagtgcctgccac gaaacagcacccagaagtgcaggtggaagatactgatgctgatagctgccccctcatggcagaagagaatccaccctcaaccgtgtt gccgccaccgtctcctgccaaatcagacacccttatagtcccaagctcagcctcggggacacacaggttaccttttatgcttctcttggg cttttctcggcactgtgccacctgtttatttggcagatgaagagactgaggccagagggagaagcagctggactaaaatcacacagcgagttggtgtcagaagtgggactatgatgccccttcctcgttttcctaactcctgaattaggtctcttataaaagctctcatgggaggccctgt cccaaatgccctcagctataagttagaaggagtaatataataaataatatagtaatagcgaggcacggtggctactgcctgtaatgccag cgctttgcgaggctgaggtgggtggatcgtttgaggccaggagtttaagaccagcctggccaacatagcgaaaccccgtctttcctaa aaatacaaaaaaattagctaggcatagtggctcactcctgtaatcccagctactcgggaggctgaggcatgacaattgcttgagcctgg gatgtggcagtgaactgagattgtgccattgcactgcagcctgggcaacagagcaagactctgtctcaaaaaataaaaaaaaaaaact ttttaaatttaaaataaataaataatatagtaatacataataaatgttacagaattaatttctacctccacattttattttatttcatttttgataggat gctatttattaaaacttgcatattataactcaccaagtcttttttaaaaaaaaaactcttaaattttaagtgtacagtacagtattgttacctatat gcacattatttacagcacatctctagaatttttcgtcttgcatgactgaagctctatactcattgaacagcaactccctattcttcccttcaccc ccaccccagccccggcagccaccattctactttttcattctactttctgtttctatgagcttgactactttagatatctcatatatgtggaatcat gcaatatttatctttttgggactagcttatttctcttggcacaatgtccttaagattcatccacattgtagcatgtgacaggattttcttcctttttt aaggctgagtaatgtcccattgtatggatatgccacattttcttttctttgttcttttattttcttttcttttttccttcctttcttccccctcccccttcc cctcctccttcccctctctctcccctcccctctcttcccctctccttccctcccctctcctgctttcttcctttcagacaagatctctctctgtcac ccaggttggagtgcagtggtgtgatattggtccactgcagcctcaacctcctaggctcaagcatttccccttctcagcttctcgagtagct gtgactacaggcacatgccaccacccagctaattttttattttttgtagagatggggtcttagtttgttgccaaaggctggtctcaaactcct gagctcgagcaatccacctgccttagcctcccaaagtgccggaattacaggtgtgagctgccaacacagcccacattttctttatttatct gttgatggacatttatattgtttccatctcttggccattgtgaatagtgttacaatgaacatgggagtgcaaatatctctttgagatcctgatttt gattattttggataaacaaccagaacagggactgctggatcatatggcagtcctatttataatattttgaggaaccaccattctgttttccac agtgactataccatttttcctatttctccacatacttgtcaatgcttgttattttctgtcttatttatttatttattttttggatcatggatatcttaacag gtgtgagatggtatctcattgtggttttgatttgcatttctctgattatatggtttggctctgtgtccccacccaaatctcctctcgaattttaatc cccatgtgtcaagggacggacctggtgggaggtgaatggatcatgggactggttttccccatgctgttctcatgacagtgagttctcacg agatccaatggtttaaaactgtggcacttctggcatttaaaactgtgtggcccctccgcatttgctctttctttctctcctgctctgtcatggga agatgtacctgtttccatctgccttctgccatgattgtttgtttcctgaggcctcccagttatatttcctgttaagcctgtggtactgtgagtcaa ttaaacctcttttccttataaattaaaaagaaaaagtctctctctctctccctctctctcttttcctctctccctctctctctctctctctcactgctg tgtaagatgtgccttgcttcccctttgctttcctccatgattgtaagtttcctgaggcttccccagccacgcggaactgtgagtcacttaaac ctctttcctttgtaaattatccagtctcaggtagtcctttatagcagtgtgaaaatggactaatacacccaacgattagtgatgtcgagtagct tttcatattcctgccagccatttgtatgtcttctttggagaaatgtctattcgagttctttgcctattttttagttaggtaatgtgtttgtttgttttcat attgaggtgtaggagtttcttttatttatcaaatatataattttcaaagattttctcccattctgcaggctttcttttcactgttgtttccttctatgca gaagtttttagatggagataatcccacttgtctatgttttgcttttgttacctgtgctttcagtgtcatctacccattgtattattagacaatataat gtgggctgggcatagtggctcatgcctgtaatcccagcactttgggaggcggaggcagatggatcacctgaggtcaggggtttgaga ccagcctggccaacatggaaaaaccctgtctctactaaaaaatacaaaaattagctgggcatggtggcacatgcctatagtcccagcta ctaaggaggctgaggcaggagaatcgtttgaacctgcgaggcagcggttgcagtgagccaagatcatgccattgcattccagcctgg gtgacagagtgagactctgtctgaagaaaaaaaaaagataatagataatataatggaaacaccacagaacactagaaaaataatcatct acaatcccaccacccaagatggtgtttttactactctgcattctcctctatgccttggttaggcccatggtcttgaatggttttaattggagga gggatccagtttacggtgctgtctcattccatgaacccttttccatattgctgggctgtctccttaatgctcacttatgatggctgttgggtatt cctccagctccatgtgccggaatgaacttaacaaatcccctcttgttgggccctatccctaatggcaccatgatcacgtgtttggtcttccg ctctctgcccttaggctcctttctcaagagtgacactcctagtcaaggtggtgcacatttttacacctcttcagataaatttgttgctgagtgg ttagagctgcttttggacaactcaagccaggtgcctgggagaaggctgtgtgtacagctggaggcaaactgccaagggcaggacagt gctttataagctaagggaggctgtctgtaactcttctgcactggttggatgtagagatggttctttcttcttagttaatttttgctctcatccatg agctcctgagtaattgctcagctgaggtcagaaggctgtcagagtggagatggctaggtgtagcaaactgagggctgtgaggtcttaa cacagagaaaagagacacagtttcactgagccttggagtacagctcacaacttcatcagttcttatgtttagttcactgaagtatgtacaa agcccattaggggatccagaaatggctccacttgtagacatctctccatttacccctgttccctttatatgggcagggaaacaatgaagg ctgggctgggtagggggtggcatatttaggggagggaggggaccaagatcatgtgtggctggaatctaagcctctggctttctgcttgt gagctgggcctgtgaggctcctcttgtccctgtcattcatgcattcgcaagcatgtactgaacatctgtatgtgctgggctctgccctggg tgctggggatacatgagggagaagcctggcaggggcctgccctcatgaggctgacattctagaggggccaagatagacaatgaaca agtaaccaaataaacaagcaagtgccaggttgactgtgactatgagaaacagggtgggaaacctctgcctgagcccagccctgaga ggggagagttctgaggcctgcctctctgtagaggccgagggcggtggggctgactgtgagcagggacagaatgaacattgggagc caaaaagccacagaggagcttgagcaagtccctctttccagcttaaggccatggcttggagtgaatcccagagcatcacttgtaagac gtgggtcttaaggcaggtcatttaacctccctaggcctccattttccccgtgtaagacgaggaaagaaacaccacctggttactagaatt gtgagaatttaacgagctactgcaccctaaggtactgagcacagggccagacacaggcagtgctgggggaccagaagctgccagct taggtcctcattctggcttctctggggcagcttgacttttgtgggttttgttttgttttattttgttttgttttgttttgagatagagttgtgctcttgttgcctgttgcccaggcaggaatgcaatggtctcagctcactgcaacctctgcctccctgatttaagcgattctcctacctcagcctcccaa gtagctgggattataggtgcctgccaccatacccggctaatttttgtatttttagtagagacgaggtttcaccatgttgggcaggctggtctt gaactcctgacctcaagtgatctgcccgcctcagcctcccaaagtgttgggattacgggtgtaagccacagcatccagcggcagtgtg cactttctaggctcaggagagtgccctgtgctgggcacaggaggatggactttgggtcccagcaatggggtttttggccccaatcttctt gtgtggggctgttcttgttaattacagtgtgaaaaaagtgaatccatgaattttgccatcctcggcgagttggcagaaggcaacaggatc atttcagagccttctggaatcaaggtggatgtcacacatccctccctctggatccccctagaaatcctagtggtacaatcctgagtcaaga tgatgaggctggggaagagcgctggcctagaaccctggcagactcaggtttgggtcccagtggagtgctgcgtctgcactgtgatgg cctcagtgacccctgccccagtgtcctcctgcccagctctctgattggccctcaggggctggagtaggaacctctgctgaggtgagctt cttgggaagatgtgcagatgtcagcaggtttgggctgggccccagggtgtgttctcagaggctcctcctctgacactccagctgggga aagggtggaatttggggactggctggagttttcagcttctgctgggaattgcagattaatcactaaagatccttgtcaggcttggtcccaa ggtgaagctccctgcctgtggattccatttctgatcaaacctcacggaaatgaaggccctggtaaaaagtaatcatggccgggggcgg tggctcatgccggtaatcccagcactttgggaggctgaggtaggcagatcacctgaggtcaggagttcgagaccagcctggccaata tggtgaaaccctgtctctactaaaaatacgaaaattagctgggcatggtggcacctgcctgtaatctcagctactcaagaggctgaggc aggagaattgcttgaacccgggaggcagaggttgcagtgagctgagcttgcgccactgcattccagccagggcaacagagcgaga ctccatctcaaaaaaaaaaaagtaatcatgaacttcccttgggaaaccaaggtcccaggacctattttctggaagtcagctccaccatttt actttcacagagtcacagctaaagaccgaagcaaggctgccctgtgttcaccaaggctgaggagagccctgctgagtgtagcaattgc agcctgaccttcacagcaggtgcccagcctctaaccgttttctagtcctaccactcccaacttgcattttattttcatgtccccacagccaa gtttctcttgctgattcagctgtgtgtactaccatgttatgcccatttttggtacactcaggttcaattcctgtattagtctattcttgcattgttgt aaagaaatacccgagactgggtaatttataaagaaaagagagaggccgggcgtggtggctcatgcctgttatcccagcactttggaag gctgaggcaggtggatcacctgaggttgggagattgagactagcctggccaacatggtgaaactccatctctactaaaaaaaaaaaaa attagctggtgtggtggtgtacacctataatcccagctacttgggaggctgaggcatgagaattgcttgaacctgggagacggaggttg cagtgagccgagatcgtgccactgcactccagcctgggtgacagagcaagactccatctcaaatgaataaataaataaaatactaaaa aataaagaaaagaggtttaattgggtcattgttccacagactgtacaggcagcatggcagcatctacttctggggaggcctcagggag cttgtactcatggtggaaggcaaaagaaacaggctctcacatggcaggagcagaactgagagagagtggggaggtgccacacagtt ttaaacaatcagatttcatgagaattcactcactatacagtaccaaggggggatgttgccaaaccattcatgagaactccgcccccgtga tctaatcacgtccaccaagccccaccgccaacactggggattacaattcaacatgagatttggtgcggtcacagagccaaaaccacgt caagtcctaaactctcatcttcatgctactgactgatctgatgctcatcctgatgtgctttcatgtgtgacatgcatacccatgtccccagact cttggtttcatgtgctaacactcagccttctgtgttttaaatgctaatggaagtcattaataatgaccattcttctccaccaggacttcattcat ggacatatgaaatgctcttcctattggtagatattggcctgggcggagtcctccagacacaatctttgtgtttctacatcagttagttctactc tatcttaaagaggattctttctctaacactggagcttggtttcttggtggtattaaggaatttctctctcttgatcttttttcctctttcccaggcttt tgcgtttctcagtaaaccaattcattctctctgctgggacagagggtgggaaaatccagagagttagcccctaagataccaacataaaat agtgtcaaaatcacctcttccaccaagtcagacacccaggctccaacctgctgatgtcaggccagaggcaggttgggcaagaaccttc ttgtctctgaaattcaggtggatttcagatctagacaccataagccaagacactaatggctcaagaatgaggtactccatggcagacttg gacttcctgactgatggaggaccccaggatcatttagtcctcattcttcagatagggagcctggggctcaggaggggatgggacttacc taaggtcacaccgtaaggcggagactggattccatggcttctctgatagctgaaaaggtggtgtgatagagagtagaggggaatgag gtgggtgactttagacagggagcaggagaggggaaggaagttgaaactgagaactaagtgataagtatctcatttacttgagtagattt aatcccattttacagagggggaaactgaggtaactcagcatgttttggtaatttgctgaggccacacacaggtaagaggcaggactgg ggtgcaataccaatgctgttacctttaggccaagccatttaacagatgggaagactgaggtctagagagtggctatgcccctctcttctg ccgttttccctccctccaggcaaatgaggactcagaagttcatgctccgccccccatcctctatctaggaattggctctagatgatctttag ggatcaaagtcatgacacttctccctggggcctctgcctttgctgaaggccccttccacatctgtgtctttgcaggaagaagctgccctct gaggatgatgaggctgaagagctcaaggcgttgtcaccagcagagtccccagtggttgcctggtctgaccccaccaccccgaagga cactgagtgagacccctctggggtctcagaaaattgaaagactgcggggagggtgggggatgtgggggggctagtgtggatgctttt agaaccttctaccgcaaggcagactttgccagtggcttccgagcagtaagtcgggatcggtcgcctgcccaggctgcttctggctggc ccaggagtcctgttaaatccagtgggctcaacctctgggcccagctgtgtccacggccagcagtgctcagagagggagggatctcta ggcagaggtccccttgtgaaggagaggctgaggggagctgcctgcagggctcctgctgctgaacttctgaaaggtcagtggggaca gggaggaccaccaggatctccagtgggttctgccctggggagtttgtgaaaaaagaaaaaaaaaaaagattccctggccctacctgg ctgcgctggatcagagctgcagacagtaggcctggggagttcacattttccaaactccccaggggatgtggtgcagctggcctgcag aacaatgtttagaaaccactatgatgacggccaggcgtggtggctcatgcctgtaatcccagcactttggaaggctgaggtgggcgga tcacctgatgtcaggagttcgagactagcctgaccaatatggtgaaaccctgtctctaataaaaatacaaaaattagccgggcgtggtg gcacatgcctgtaatcccagctacctgggaggctgagacaggagaatcacttgaacctgggaggtagaggttgcagtgagctgagatcacacctctgcactccagcctgggtgacaagagcaatacttcatctcaaaaaaaaaaaaaagaaagaaagaaagaaagagagaaag aaaccactgtgatggcattgggacctgggtctctgagatcgaactgtccccagggctgggccagtgggaggtttcttcccgaaggatc tgagaagctctggtggatggcgaatacccgggccctgggctctgcctggaatccccaatcacagtccttggccatgactcctagatga gccacaggagacagtaggctttccttcttccttccagtctttttttctatccacaattttatgcagtttatgcttaacagagctgtggaaagtta gaaaagataagcaagaagaagccccaaccccaccctcagtccagttgcccactgcagcacgcagtggactcacattttatgagatag ggtaggggtagggtttaatatcagaaacgaaggcagaaatgaggtataatgagaaaacctattcttgaaaactcttaaggaggtcacct tggagacccatgtattagctaggcaggctaggctgctggacctcaaatggacctcaaaagctgagtggcttaaggaactgtatcatata ccatcatcatctcaatagttgcatgcatcaggtcagcagaaggttctgctccatgcaatcatttagggacccagatgacagtgctctgcc actttatccatgtgccttcccatgtcccactggtcaccactagaaagggagaactccaggcctgtcttcagtaaggaggcccctagttgc atgtattgtgtctgagagctcacattccaatagcgagagcctggcacgtgatccctggctgggtggctatatcccagtgagtgtgcacca caggagagaatgagtgtaggggagcagctcatgtccccgccacacctgtatgccagctctcattaggagccccacacttggccgggc acggtgtctcacgcctgcaatcccagcactttcagaggccgaagcaggtggatcacctgaggttgggagtttgagaccagcctggcc aacttggtgaaaccccctctctactaaaactacaaaaattagtcaggcttggtggcgtgcacctgtaatcccagctactcaggaggttga ggcaggagaattgcttgaacccaggaggtgaatgttgactgagccgaggtcataccactgccactccagctgggcgacaagagtga aacttcatctcaaaaaaaaaaaaaaaaaaaaaagagcctcattttctccagcccgtcttcctcagttgagctcaaaaagagtagtcgact gcagtttctgtgtcctaatgtatgaaaaaaaaaaagataaaaaagatattcagctcagcaagatgatctcagtgaggctactcagaggaa agaaagcagtttctctctctctcttgctgacagtgcagaaactttaattcctgtaagttaaagggatatagaatgtatctgcactgctttttcc accagttacatcctaagcctaaaaaagtaggcctgctccctctctgttctgcccctggacacatatctcatgtgaaccctgcactgggatg aacagcccaattttgctgaactctaaactgagttccaagagtttcctcctaaattacaagtggagtcacgctttttccagaactgcccatct gaatcagagcctacgtggtacattttccaaaagctgggccagaacacctgggcattattcccggactgaatttccttctaggtgattacg gtaggtctgtggtttctgatgtcagaaggagatgggggcaggacttacctccttcaggctctgtcagaaggtgggacacagacgctgc acagctctggcctgggggtgcacaggtccctcccggctcagcctgctcagctttgggtgatgttcccacagtggccaggaccgtgcg gcctccacggccagcacaaatagcgccatcatcaacgaccggctccaggagctggtgaagctcttcaaggagcggacagagaaag tgaaggagaaactcattgaccctgacgtcacctctgatgaggagagccccaagccctgtgagtctggagctggagggggtctacctg gatggggatggggtggggcctccaaagttccctgtcaactcatggcaagggtggccccaaggaaatggttgaggtccatgagagaa gcccagatccccactctctggatttgggatccatatgtacaggttgagatatgtacccaagggtgggaagatcaaagttctgaccaaca atccacttgcactcggccctctctcccatggggcaagtgccttccttgacccttctgtccctttcctactggtaggtgagtggtgccaccc caaaatgtatcttatggtcttacactcagattgtgtgcacagcctgctctggaactaggcagtcctatttttcagagatgtcagtgccttgaa agttcttattgaggctatccgaatttttcttttctttcaaggtatcaggaggaaataagacacatgatctcttgtcagggagattttttagatctt tccagaggaaggctgtatcttggaaatcatggtgcccacaacaggccatgcagagcctgatgtcccagtcttgaagctggaaaacata gccatattgaacctataagagccaatccctagaccaaaagtatcatcatagatgcacagggaggctgtgatcccatctggtctctgttttt agagatgaaagagttgagattccctctcagaaacctggagcaatacttccttgattttgttaggcccttcagcattccagccatgggtact gggaacactgggtacaaaatgcccccatcccatcagagggaacaaagggggtctaagcacaacacacaagcagacattgaaacat ccaactggcccagaaacagctcatttggggcagcttctgagaagcagttccagactgtaattggtaagaacttctgggtgggcaccca ggttgctgttgctgttgaggagggttgtggagttcactccagaagagcaccttgggttctgcacacagagcttgcagtagcttgtataatg gtgccagggtcctctcagatctggatccaaggtctgcagttgggacccttctctctgcatcctctacctgggcctcagggctggttcaga tggtcaccaatgtcatcccacagccccagccaagaaagccccagagccagctccagacacaaagcccgctgaagccgagccagtg gaagaggagcactattgcgacatgctctgctgcaagttcaaacaccgcccctggaagaagtaccagtttccccagagcattgacccgc tgaccagtgagtcctggggaatgggtgttcccactgtgtaagatgagagagcaggagagttggagcgggatggggagagggtcag gggattcaagaggtgggatgggaggccacagacaacctgggttcatgatcttttatacctttcaaaagtattccacagtttcttgcctgatt ctcaaaacaaacccagcctggctaacatgacgaaaccctgcctctattaaagatacaaaaattagctgggtgtggtggtgcacacctgt aatcccagctacttgagaggctgaagcaggagaatcatttgaatccaggaggtggagagtgcactgagtcaagatcgcacaattgcat tccaacctaggtgacagagtgagactctgtcttaaaaacaacaacaaaagtccctggaataggaacataagatcatacccattttacag atgaggaaactaaggtacagataagtcagccagggtaaagatgcaactagttagtgcggacccaggactcctgactcttggccaagtc ctttttcactctggtcgtgctggccttttcctagagctcctgctctgaacagaaaaagagcaggaatgtgaattcctcttgcttccctttaag cccacttcatattcctcagaaataacccacttcatatgtgcatagcactttagagtttacagagggctccctgcctttcttatctgaccttttcc tgtgatttctcagaagagggcaggtgccagagagggactttctcaaggtcacacatttggggggcaggacttcaaccctggtgtccact ggattctttaaataacctgatgggctgggtacattggctcacacctgtaatcccagcactgtgagaggctgtagtgaaagaactgcttga gcccaggagttcaagatcagcctgggcaacatagtgagacctcatctctacaaaaatagtaaataaataaataaataaataaataaataa ataaataaccagatgatctggaatacccatctatccatccatccatccatccatccatccatccatccatccatccatgcattcacctgtctgtccatctatctgtccatccttcatgcattatttactattctgtatttactgaggaagaaaatacattttgggaaaaaattaaagaagaataaaag caaatgagttttccagttggctgcctattatcttataataccagcatagtccctcctgagagtactctgatttattagggattagcagtaattta tttttaaaacagatagttgtgtttttttttttaagtagagatgggggtcttgctatgttgtccaggctggtcttaaactcctgggctcaagcaatc ctcccacctaggcctcccaaagtgctgggattacagacatgagccactgtgcctggctcctaaagcagagagttgtgtgtgtgtgtatgt gtgtgtgtgtgtgtgtgtgtatatatacacacacacacacatacacacactcttacaaaagtgccacatgtagtagttgaaggagtagaaa gaacaaaaccctacctgatccaactggtgaagagccttgcctggctgatgctctagcgtccccgtcaaccttccccacagacctgatgt atgtcctatggctgttcttcgtggtgatggcctggaattggaactgttggctgattcccgtgcgctgggccttcccctaccagaccccgga caacatccaccactggctgctgatggattacctatgcgacctcatctacttcctggacatcaccgtgttccagacacgcctgcagtttgtc agaggcggggacatcattgtgagtcacaggcaggtggtgttggggtgaccgggggcagaccgctcagccagagcctccagggca ctgacccctgatgctgtcatagatgacaaaataagtgaataaggaaatctcaagaacggctgctgtgcttgttcaatagcatggttgtttat tttgcatgcctttgatacgttcaaattacgcacatcagatattgctcataggtgaaatagttcctggaccacttgacagaatcaaagataga gaagtggtccagggaatggtctttagtgaggatgtacctggtttgaatcccacctccaccacaactagctgaatcgcctctgtttcctcat ctgtagaatggggctagtaatagcttccacctcccagtgttgccatgaggattggttgaaacgatgcatgcggggcgctcagcacaggt atccactggcaattgacttctcccaggggtcctgcaaaaacacactgatgctctcctacttgggccaatttgagaccctggcctctggtc cagccccgtgcaccacccctgccttaccagggaggaaactgaggccctgactgtgacttctcaggaacaggagacacagtccctgtc tcccagtgtgggtcactttccaaaattccaccccggcagtgcctgttttgctgtgtgcagacaagctcaggttttgcctgtaaacgattttc cttggagagatttaaagagagtaaagtaaaaagtaaagagatttccttggagaacctcttttctggcctgatcagagtgactaccaattcc ttagtaaacagatgtgcaaatggtccaggtaagaccaatgggcagagggaataagcttaatgtccttgctttccacccaagaaatcagg atctagctgggtgaccagaacagatccctcctccctgggggctgggcctctccacccataaaactcaaggctttcaccaaggctcaga tggtccttgggctgttatgctcgtttcagggtggggttcttggggaagtttcacaattgctccacagggtcagagattaagctgtaaggtc atttcaccatcatcattccctttctttcagacggacaaaaaggacatgcgaaataactacctgaagtctcgccgcttcaaggtgtgtgccg ggcatgcaaggtttagcgtctgcggggtttcagaagggggtgcctcagagggccctgggtgcctggggcgttcatgagatgtggtttc tgccacacggagggatgctggtccctgtgggagtgtcctccaaaccacaagccccaggagaccagcacaggctgggtagccccca gaagttatctttcttctttttcttttcttttcttttttttttttttttgagacagggtctaactctgtctcccaggctggagttcagtggtgtgatctcgg ctcactgcaacctctgcctcctcgttcaagcactcctcctgcctcagcctccctagtagctgggattacaagtgtgcgtcaccacaccca gctaatttttgtatttttagaagagacagggtttcaccatgttgggcagtctggtctcgaactcctgacatcaagtgatctgcctgccttggc ctcccaaactgctgggattacaggcataagccaccgtgcctggccagaagccacctttcattgtgtctgtaccactcccgccccttactg tatggcacagcagccactatcgaggataatgaagctgcccttggggctggcaacacgggctttgaagccaaacaagggcttgtcggt gggacaccttctgccctctagttttaagctctactggggcattctgacctcacctttccatggaagggagagactccgcctctcactcccc tcaccttccaccctggagtcatccgttttacccccatggtgtcccactgggatttcagatggacctgctcagcctcctgcccttggattttct ctatttgaaagtcggtgtgaaccccctcctccgcctgccccgctgtttaaaggtaggactgctgggcccaccaccccaggccttcagct catccctgctctgcaccgacactggcatcttcgtgtctgcccccagcagcactggggaggggagttcctcaagggatggcccgaggg gagatgcatttattccacgtgtgtcttagtggaaagttcattacaatttgctagtgcttctgtggtgctgcatgttaggtagtattattgccagc attttacaaatgaaggactggggcacagagaggttaattaatttgcccaaagtcacacagccaggagggagtggagcccagatatga atgcaggtcatctgtctccactgtttacgctctcagtgattatagagactctcaagaggggatcttctgctttatgatggtctctgtctaaaag atggggacattggtgcttggagaggctgggggacttgccctaggtcgcaggagcaaggaagtgccagggatggatttgaatgcaca gccagacacccgtttcatcactggccaggcctcagtgtcattccagccattccttctggcactcactgggcagagcgaaagtctccagg atggtctggctgccacagctggccatggtcctgtcacgtgatcagggtggatagccactcagagcattcaggatcccccaaatggattt tgatggccaaccaatcacattcatctgtctcctgaaatgaagactggtgcctttttctgaaaacagtactgagggaatttacaaaaaatgc aaatgcaataacatgaacaacccagagaaacagggcagagaaaactagggggcgtgcaagtgccctgatgagaagaaagcacac ctatgtggaatcgagcatggaagctgcctctgagcttcctggctgccaagaggaagggggaaagtggaattggcaccattcttatttga cacataccagcctcccaggggctgcagaggttgctttagagccaaacatcagagtgaggaaggttgggtccacaggccaagtttcac cccagacgcagaggtgagatccataccttgggtcacatggtggccccagtgagggggcaaaactcacaggcctaacattgaacacc attcagtggacaaaactctttaagtgagccatccaagcccctgttctttctgcctccagtttaaactgtggctggaactagaatgaacacat ggatgataactggaatatcccttagagaatcttcataaatactattttcaaatatcaaaaaataaatccaaagatctagaggtggtatagcat aaggagagagcttgcagctctgagtctggcctgggttggaatgctgactctctcacataccagctgtgtgctttggacaagttactggac ttccctgagcaccaatttctgtcaaaggagaatgataatacatacttcatatgtaaataaggaataaggtaatgtatataaatgtatgtgctg gctgggtgtggcggctcatgcttgtaataccagcactttaagaggctgaggcaggaggatcacttgagcctagaaatgtgagaccaac ctgggtaacacagtgagactctgtctctacaaaaaatttaaaaattacccatgagtgttgatgtgtgcctgtagtcccagctacttaggag gctgaggctgaaaaatcgcttgagcccagaagttcgaggctgtggttatctatgactgtaccactgcactccagcctgagtgacagagtgagactctgtctctgaaaaaacaaaacaaaaaaaggcatatgccagtcatataataggtgctcaataaagaaaagtagttattaatatact tttattcttataaataaacctttgaaggatataagagtggggtgttagggatgctgtctctttattaaattgttttctttataagaaacattaaaaa cagcagctttaacctggatgcatgtacccctaaaagtctcatgatagaattcagagggtccaggagtcccctgaaattatatgcaaattgt gagaacaagattacatgtcatttccaagggaagacgggctatgactttcttcagattcctaaatggagtttctggcccctacccgtcaagg ggctgaggacttctgatattgaatgctcaccgtcctgttccctcctgggggctgcaggaagagccctgagtctcggcgcccgtgggcg tgggagcagcatgctacctatggccggtgggctgctctcacgccctcctctccctccagtacatggccttcttcgagtttaacagccgcc tggaatccatcctcagcaaagcctacgtgtacaggtgagactccccctgcatgtgcatgcatgcacacacacatgacactcacaaaca cacaacacacgcaatacacacacaacacagatgacacacacacaacacacacacgacacacacaacacacacacaacatacacac atacatgcacacacaacattcaacatacacacacccaacacacaatgcacacacacaacacacaacacatacatacgtgcacaacac acacacaacagagatataacacaaaacaacacacacaaacacacacaacacacaacacatgcagtgcacacacaacacacaatgc acacacgacacaaatgacacacacaacacacacacacacacacacaacacaggaagtttgcacctagaacagacagcaagacatt gtctacccccactgacctccaggccccgtctccactggaagccaggcatcttcctggtcccgcctcctctttcccagagaaatgtttgtta tttagagaaccaggaatatccgatgcccctcattgtgatgagggtgcacatcagctggcacgaggggcttaccagcattgcagcgaac cgggcctcattttggcctcagctcagggaactggagcccccagccagctctaacagaatgggaacaggtgtaaggtggcggtggct gtttcacagatgtctgagcatgaggtgcaagaccagccaggccgtgctagggagcaaggctcctgtccacatctctcctggggcgac atggccccatcttcaactcaactcctctgcacacatcagtccctccctgctcctccacacacacacacacatttcccctctggcccctcca tcccccagtgcacagtcagctctgctacctctgttccaatcctagccccaggtcatgacaacgccccattcaagcaccacccttcctaac tggtgcaatctcaacaaagcatccagtcgggccagcccagccatgggttggctctcactgtccaggtgtctatacctcacctcccccttc cgccgctgtgttcatgcgctgtgttccttctgcagggtcatcaggaccacagcctaccttctctacagcctgcatttgaattcctgtctttatt actgggcatcggcctatcagggcctcggctccactcactgggtttacgatggcgtgggaaacaggtgagccacagtcctccccctgg ggccatgcctgggtgaccttcctgttctttataattcgcagcacagagactccaggaaggagtcttattgctgctgtattgccaaggccac tttgctcctaagcaatcactggcatgacaaggctcaagagccacatcttgggccgggtgtggtggctcacacctttaaccccagcacttt gggaggccaaggcaggcggatcacctgatatcaggagttagagaccagcctggccaacatggtaaaaccccatctctactaaaaata caaaacttagctgggcgtggtgatgcacacctgtaatcccagctactgggaaggctgaggcaggagaatctcttgaacccaggaggc ggaggttgcaatgagccgagatcgtgccattgcattccagcctgacacaagagcgaaactccatctcaaaaaataaaaatagaaaaa gagccaggtcactacatcttccccaagctaaggtttcctgagcacctgcatgtgccaggccggaggtactggtgtgaacaagacttgct tggatcctttctgcatccaaactctattagggtgtgcaaatacgctgtatttgtgcatctactcctgcttccccgggcatggccaagaggga atgaggccagagaatactccccagcacagctccccaggcactgctaccatggaggtgctggcgaatgaggtgagacccacttgcca atcagagagccaccatccattatgagacttgccaaagggatgatgatgacgtgcgaaccaaaaagaatctggcacaagtttcaatcaat ttagaaagtttattttgccaaagttaaggacacgcccgtggtacagcctcaggggttcctgatgacatgtgtgcaaggtggttggggtgc agcttgcttttatacattttagcgggacaggatacatcaatcaatacatgtaaggtttatattggttagatctggaagggtgggacaactca aagggaggagtggcttccaggtcataggtagatttaaaaaaatctctgcttgggaaaaaaaaaaaaaaaaaaaaaaggccgggcttg gtggctcacgcctgtaatcccaacactttgggaggctgaggcgggcagatcacttgaggtcaggagttcgagaccagcctggccgac acagcgaaatcccatctgtaataaaaatacaaaaattagccaggcatggtggcagcgcctgtagtctcagttactcgggaggctgagg cacaagaatcacttgaacccgggaggtggaggttgtggtgagatgaaattgcaccattacactccagcctgggtgatagagcaagact ctgtctcaaaataataataacaataataaataaaaattccctgcttggcaattggttgaaagagttatcagtagaaagaaatgtctgggttat gataaggggctgtggtgacctaggtattttcatgcagatgaagcctctaaatagcgggctttagagaaaagagactgtgatgtttcttacc agacttaaggtctgtgttgatgttaacgctggaggggtacaatgaggcgtgtccgaccccctcttccttcatggcctgaattaggttttca ggtaactcgggaaagcccttggccaagaggaggggtccgttccgatggttgggggccttataatttatttttggtttacaatggtggtgat ggtgacaatgatgatgataatagcagcaatgcttgtaggatacctattttgtgccagccctgcttaaagcactgaacacagtaattcacaa ccatcacatcagatagtaccgccccattcccattttacagctgaggtagcagatgggagttcccaaggttacacggttcatgagttacag agctgggatttgaacccaggtgtcatggccacagagtcctaactgcctcgatgaaaaggaaaagtgccaacctggtggccagaggg agtgggggaggtgggcatgagcactgttgattccaccaaaagtcaccaaaagttactaacatttaaaaaaacttttttagccagatacag tgactcatgcctgtaatctcagcactttgggaggccaaggtgggcagatcacatgaggccaagagtttgagaccagcctagtcgacat ggcgaaaccacatctctactaaaaatacaaaaattagccaggcgtggtgatgcacgcctgtaatcccagctacttgggaggctgaagc aggagaagtatcgctggaacccggaagggggaggttgcagtgagccaaaattgcgccactgtactccagcctgggcaacagagag caggaaaaaaaagtttagaaaattattttaatgaacagacttaatttatttaaaagtggtttgaagtttatagagaggtccactggaaagtag agagagttcccatatatatgccacctccattattattattgttactattttttgagacagggacttactctctgttgctcagactggagtacagc agcatgatctcagctcattgcaacctacacctcatgggctcaagtgatccccccacctcagtctcccgagtagctgggaccacaggca cacgccactgcacctagctaattttgtattttttggatatatggagttttgccgtgtggcccaggctggtctcggcttcctgagctcaagctatctgcctgtgattggcctcccaaagtgctagagttacaggtgtgcacctcattttaaggctggatggtgttgaaggcgacactcgctccc ctgccctgtcaaaagccttaagcaggagagtgagcagagggagggctcagccctcgtgctgatggaagagtgcactgggggtgag ggcaggcagcagacagccagcaaggggccctgctgtgtcagagagagcagcaaaccctccaggccaaccggtggcttgacctag gggagggagggcagtgtgggcaggagggatgcgcacaggatgggggctgggcctgcagagtcagagcggaggcttgagggac gagggatgaggagtccccaagtgtgactcctagagctgccccctggcagcggagacaggccacttctcttttctgggcctcagttccc tcacctgcaaaatggggctgcaaatgggactcccaccctaaagaagtgacaggagtagatgcctctgagcccctgcctcagtgcctg ggcacagtgggcgctcagtcagccatagtgaggatgagaaggtggaagaaggaaaggtccagcggggagttactcaccgaaggg tgcccgcctcaccctgggctgtgcccactagtgacacaaaatgcccgtggtccagctgtctacccccaggcccttgctcccagtggga acaggtgggatgcttgcgtccaaggtggcggctttgtagagtggttaattgttcagaacgtcccctcccagtatagatgccccaacctca gccatcatggctatggccgtcgccttctgccatgagccctgccccgagggggagccacgcctttacagagatgtcccaggctggtttt ggtggctcatatctgtaatcccggcacttttggaggctgaggcaggcagactgcttgagctcacaagttcgagaccagcctaggcaat atggggagaccccctctctacaaaaaaaatacaaaacttagttgggcatggtggtacacgctcgtaattccagctactcaagaggctga gatgggaagatcgctggaacccaggaggtggaggttgcagtgagcccagatcgctgcactccagcctgggtgacagagtgagacc ttgtctcgaaagagatgtgccagaggccagggcagcccaggcaggcaggagaggagtccctgagagccactgtgaggtggagaat atcagggatggggtcaagatctaatgcttccccacgtcaccagtggcccttttacccattcccattcacagccctgctcctgagccctgg ctaaaccatgcattcattcacttccttattcatctaacccatcacaaggaggcagagggggacttgaagcccctttctatgcaaagggga gaatgcaatttctgcatggaattgaactctgtgagcttaccctttctgagacccagtgttcccacaggcacaatgtgggtaacttacatcca gatttcttatgagaaataataaattaatatgtgtagcatgtctagcctgggcctgaacaggagtcagtggcccctgtgcctagctatggct attgttagggggaagccagcatggcaggaaaggagttaataacctggggtatagagattttagggggtcacaaaccccccagacatt ggagctaaaatttgagcatatgttggcttttctagggaaagagagctggctttctctagattctcaaaggggcttgtgccccagtaaacta agaactctcctccctggggctaagcctggcacatccgtcactgggcagctcagggaagggagagtaagcagagcagggcaggggt ggggcagggggacagctatctgggacctccttctgcagggccctctgcgggaggagctcgagctgcccagcagtgggaagggca cgcttccttggcagcaatactccttcccaaatgtcgtcctgcagggaaccccaacatgtgaaccgagtacgagtaatgaaggaggattc ttaagaaattccaatagcatgaagaataacaacttttacaaagcgaacatttgtggagtgcttgctgtgtgccaggggctcagggtcccg agctcctacaaatgctaggccctgcatcccgccccagtgctctcctgccccagttctgcccatcctgaagcacagatggtccccacag acaaggacaaggagacttcagggaggcccctaagatgtggcatgtcattggaagattcctccagctaggaagggtttggggtaaagc cagagctttgtggacatcgcgcctatgtcctctctctgttggtggcccagcggcataagcacccagctcccggacagccgaggggtct gcagagttgggggtgctgctccgtcccgctaaacgctgcttcgcgtgccctctgatggccagaaccagtatgacgggggcttgacag gcccgttttgggttctgaaacttgcaggcaacaggtgatgtggaggtgcagggcggcccacagccctgcccaaggccccgcagctat cagaggcagagcccctaccacactccccagcaacctctgcttcctccttcctgcacgcattgcacgtcacctgcctgtcccctcactgg accacgagcttcttgaggatatgggcgaatctgttcagttccccagggcaggcaacagtggctgtgaaggctttgacagctcccttgct gagagtccaccacgtgccaggcttcttctagcacctcattttgtcttcatggtcttctgtaagggacattctattgccatccccatcttatgga gaaggaaactgaggcttggagagagagagagagagatttgtccaagttatgcagccagaaggtggctgagccgaatggctgagcc cagccagcctggtccagagcccatgttctatctaggacactgcataaatagctgctgcaggaaagccggcagggactgcccagtgtct aggcccttctgacctcacctttgctgacccaggggagggctcagcctgaggtgtggacaggtgtcagagggacccttgctgctctgtg ataatcacacacaagcttggtgtcttcatgccttttgagctgctgtagtaaaatacatgagactgggtaacttataaagaacagaaacttatt tttcactgtgctggaagctgggaagtccaagatcaaggtgctggcatctggtgagggctgctctctgcttccaagacaatgtcttgaagg ccacagtgtccactagaggagaggaatgctgtcctcgtatggaagaaggcagaagggccaaaggaagtggaaaccctccaccaga accttttgtaacagctttaatctattcatgagggcaaagccctcgcagcctaaacggcgcccgaaggccctagctcctgacaccattgc actgggaattaagtttccaatgcctgaatttcgggggacacattcagactggagcacttggggcatttcctatgccttttaccacacagta ggacaccctcccccatacctcccttccagaaggttcacaagccgacgtcagagggttactgctggtggatgacatcttagagaagcag acagcgcccagctcttgtacttaccaactagtgaccacgggtgtgccctttccctttgggggcctcagtttcactctctggaaaatggatg ggccgggtaggggcgggtcagaaaaagtcatcctcaagatatcgtccaaacttgatgttgcttttgcttttcataagccaatatggcagg agatagcaaatgaaaagactgaaaattgaaggaacactgaaggaaattgagggagtgcggtcattaggagttggaagaccctggtct ggcctcctgccaactcttcctggaaatcccctggagcctcagtttccatatctgtaaatggggatggcaacatgtccctagagacacaac catacccgatagcatttgtgaacctgctctgcaaacggcaacgtgctctacacgtgcaggaggtggttttccttgcatagatgagctgta aagggtttctgtcttttgcacaacatgcatctatcagaggcgtctgagccagactgctcggggcggcagagtggggcctgtgatgacc catgttccctttctcctctctctccagttatattcgctgttactactttgctgtgaagaccctcatcaccatcggggggctgcctgaccccaa gacactctttgaaattgtcttccagctgctgaattatttcacgggcgtctttgctttctctgtgatgatcggacaggtatcggggcaccagcc cagcttcccaccccacctgactgcagggccccctcccactgttgccagaaatgtcaccctctgtgcccgttgcttttggattaagcctgggaaagggcactagactgagagtcaggagcctggagcactggtccctgcgcggtgaagggggtggtggtgaactcggccctattccc tctctgaggtcattatcatttgaaatgtcagttcctccgggtgcactgggctgaatggttttctgcagagaccgagggagaagcccctggt gtttctggtgtgccagggcagcatcttacattctggtcgagacatgaggtgacagtcagaggttctcaaatttatatttgaagggccccgc cccgaggaacagcacactggcttcctatttcctccagcatggagtcagacaacatggccgccgtcaccgaggaactgtattgtaccag aatcagcccagcccaaggagctttcccccatggtgtggtcccagggacctgaggccaccaaggggctggcagtctccctcccttcct cttccccacatccatgctggaggcagaacttcccgtcagtaaaactttctctgtgtgaggacatctgcctacccccctcccaacgccctg ccaggagcacatgcccgtggtctgtgtgaagtgacacatcccaaatccatggaatagcgcccacccccaatgacccagtggcttcctt gacctcccataacccttcccccagatgagagatgtggtaggggccgccaccgccggacagacctactaccgcagctgcatggacag cacggtgaagtacatgaatttctacaagatccccaagtccgtgcagaaccgcgtcaagacctggtacgagtacacctggcactcgcaa ggcatgctgggtaagatggtcacgccctggcagccaccagctcccagtcaactcctgtcgcctggtgcccggtgccatgccttcggg gcgcctccctcgtcctctgtgtctctgaggtcccgagtactggggcctgtcttggctgtgtccccaacattggtgtgagaggtgcccagt ggggcccctgaagtgcctctagcagggctgtcatgagatccctggagacggggggcttgtcttctgtgtgcctgcccccctcactttac ccaccttgaaatgcatccccatccaagagcgaagtgtagtttgtttgtgtgtttgtttatttatttatttatttgacatggagtctcgttctgtcac ccaggctggagtgcagtggagcaatcttggctcactgcaacctccatctcccgggttcaagcgattctcctacctcagcctcccaaata gctgggattacaggcatgtgccaccatgcctggctcatttttatattattagtagagacgaggtttcaccatgttggccaggctggtctcg gactcctgacctcaggtgattagccagcctcgggctcccaaagtgctgggattacaggtgtgaggccgaagtgcagtttaaattgaaa aaaaaaaaaaaaaaaggaagaaggactttctgtcccttcctccctcccttcctctctctccctccttccctctctctctcctctctcctcgcttt ctctcagggcattgcacaccccatgatgggtgcagcaggcttagccctggatctcctggggcagctctagagctccacagccaggttg gggtcctgagtttatccctcgagctacaagactacgggcatgtcactgccatttgctcatctataaagtcgatattatagtagggcacctct tgggttgcttaggggatttagtaaaatagttttgcctgtaatcccagcactttgggaggctgaggcgggaggatcacttgagcccagga gttcaagaccagcctggccaacatggtgaaaccccatctctacaaaaagtcaaaaaatttagctgggcatgttggtgcgtccctgtgag tcccagctactcaggaggctgaggctggaggatcgcttgagcctgaggaagttgaggctgcagtgagttgtgattgtgccactgcact gcactctagcctgagtgacagaacaagaccctgtctcagaaaagtaaaaataagtacctgtgcggtggctcatgcctgtaatcccagc actttgggaggctgaggcaggtggattgcttgagcccaggagttttcagaccaacttgggcaacatagtgagaccttgtctctaaaaga aaaaaattttttttagcttttcagggttcaaagtattttaaaaattttattttaataaaaactaaaactaagataatctttataaaacattctattaaa aagtgcctggcttggctgggtgcggtggctcactcctgtaatcccagcactttgggaggctcaggtgggtggatcaaatgaggtcagg agtttgagaccagcctggccaacatggtgaaaccccgtttctactaaaaatacaaaaaattagcccgcgtggtggtgggcacctgtagt cccagcaactcgggaagctgaggcaggaggatcacttgtactagggaggtggaggttgcagtgaaccaaaatcatgccactgcact ccagcctgggtgacagagcgagactgtctaaaacaacaacaacaacaacaacaaaaacataaataacaacaacaaaaaaaacaggt gcctggctcatagaagtgctccagagactttagttcttgctgatgttgtccttccagggaagtgggaacacaggtgaccaataattgcaa cagcgcaggggctactaacatagtagctgagtggaggatgtcaagggaaggcagggaaggcttcctggaggaggtgatattagtgc tgggttctgaagaatgagtaggagtttgggaaatgcacaggggtgctcctggcagaggaaacggtattggcaaatgcctggaggtga gagagggtgcactacaaatccagagtggttccatgcggcagggcaatggtgtgccctggtgagccagtaggtgggcacttggccgg tatgtatgtctggggtggggcccggccttgcctctgacccttgccaggccttgttcacagatgagtcagagctgatggtgcagcttccag acaagatgcggctggacctcgccatcgacgtgaactacaacatcgttagcaaagtcgcactctttcaggtacaccttttggtgccctcac gctcagggctgtgggaggctccaagcccaatcttctctaaaaagttcctttcccttcctcaccgggacagggctgtgaccggcagatga tctttgacatgctgaagaggcttcgctctgttgtctacctgcccaacgactatgtgtgcaagaaggtgagctggcctggcagccaccac gggatctggggttcaccatccccctggctttcaaggaactgagctggaggggtgcctgccagcccagggcagagcgagattgttgg ggagggagaagagcgtctgcggcttcatgctgcagtgacctgaccagacccaagagcaggagcggattaggaggctttgccgactc tatgcagtcaacacacatgtgcctcgggctttgcaaagcattttctcagctccattgacacaggaccatgggggccaggacaaagacc ccccccccaactccaccttacagaaagggaaactgtggagcctgaagagaggcagggccttgactgtgtagctggaactgcttctgg ggtctccatcttcccatcaggaaaatggggcaggagaagggatgggcttaggtcagaggttgcaaactggtggcctttggataagatc cagcacacaggtattctgtgggtttttttttttctaatttttaaaaaatgttgaattggtccccaacctttaaaaatttggagattttgtataaaaat tcagatttctggctgaaaacagaaaatctgccagcgctgggtccactttcccaaatggcaagaggcagctggggtggaggaaaggtt gccccaaagggggacatcgtgtcccttcctgatgccacagcccccaccactccccactgccccaccctagaaccccctccctgctca gcttcaccctctgatcccagaaaacatctggagttgaatcctggggcctgacaatgtctagagccctttctgctccctgtccttggggttc aaggtgagaccaaggtcacttggtgagtgacgggaggggtcaggaacccaggtttcagtcctcttgcacagagccgttaataagttaa taataaaccgttaataagcatacggggggaaacattacccaagcaggctctgcaagacttgggaacccccgtccccaagaaagtccc tgtctccaggttatatgctgtgcagcagtgaaatttctggaagcagagagccggggtgaagaggggaaaagtatgcttcagagacctg agaatttaaagctgcaaccttaggggcaaagaggggtttgggagcaatagagtggcctgcctgacactggagccccttcctggctgtgcagccttgggcaagtggcttgccctctctgagtcctgctcatcttatggataaagtgggggatgttgatagtacctccctcatggggtggt tgtgagggctgaataagatgtgcatacagagcttaacattgttcctggcacacagcagctgccagtactgttgctgttgtcttcgtgggtg atgcagagtttgctttaaatgagaaaaagtgaatctagagtctgtgattgcaaattaaattgagcacttggtgagcacgtgtgaagggcg ccctctgctgtctggttttgctcggcagcgatctctctgagacaccccggggactggttactgaggtgagggtcccttcaccctttcactc ttgtcaaactaaacagaaacatggaggcaaacctctctacctttttgttaagggggtcatgcatgtgtgaaatgcacaggaaatttttgtca aataagtcaaaagtgtgacagttactgtccttggtcactccccccaccccttatttatttatttgtttgtttgtttgtttatttgagaccaagtccc actctctctcccagggtggagtgcagtggcactatctcggctcactgcaacctccgcctcctgggttcaagcatttcacctacctcagcc tcccaagtagctgggattacaggcatatgccaccatgcccggctcatttttttatttttagtcaagacagggttttgccatgttggctaggct ggtctcgaactcctgacctcaggtgatgtgcccgccttggcctcccaaagtactgggattacaggcatgagccactgcacccggcccc ctttttaattattattattattattttttttttctgtagacctcctcttccccttttaaacacatgtgactcacaggtagctggccaggacaagggc cggtgaaggcatgaggattggaaggcgcatggcagactaggtgtttctaattgcagtccaaagcaagctattcattcatgcatgcattcg gtttaacttgggattaaggccattgtattcattctccaggctgccgcgggacactgtgcaactccaccacaagctgggtggcttctctcct cagcttgtaggtggtcatctagttttttttgtgtctcctcacatcctctggtctctatgcctgtctttgtgtccacatttcccttttgttaggacacc agtcatattggattaggtctagaaggacctcatctttaggaattacatctgcaatgaccctattcccaaataaggtcacattctgagatgct ggggatgaggatttcaatatatgaatttgggagggagtacataattcaacccataacaaccatgaactctggaaccagactgtctgggtt cagatcccagctcttgggcaaagagtgatctcgactctgagggttgttgaggataagtagtacactgcataaagcaatttggggctggg cacagtggctcatgcctgtaatcccagcactttgggaggctaaggcaggcagatcacttgaggccaggagttcgagaccagcctggc caacgtggcaaaaccctgtctctactaaaaatacaaaaatttgccgggcatggtggcacatgcttgtaatcccagctactcgggaggct gaagcatgatgcctgtaatcgcagctgtttgggaggctgaagcacaagaattgcttgaacccaggaggcagaggttgcagtgagcta agatcatgccactgcactccagcctgggcgacagagcgagactccatctcaaaaaaaaaaaggcaatttgaatattatcgttattgttgtt cccattcctgttgcagttagcaaattcttcccgagcccctgagatgtgccaggtgtagacatcatttgatggtgggagctgctatctgattt gccctgcctccatcagtgagtctaactggaatttgcagaattccattatacctgcacctcccaggttagctggcacttcccattccatgaca tcactaactcccccactgcaaccctgtcagttatggagggcatggggtagtcactcctccaagtctacagatggggtgactgaggcctg agagggaagtgactgcccccacacgacaccccaagttgtggtggcagggctggaccccagtgcctaggttcctccttgccggacctc cagcctctggatccagccctggctgcttctctggtgactttgggggatgaacagagggacttgtcacttctttgtttcagggggagatcg gccgtgagatgtacatcatccaggcagggcaagtgcaggtcttgggcggccctgatgggaaatctgtgctggtgacgctgaaagctg gatctgtgtttggagaaataaggtcagacgcagctggggtaaagtgaggggcctggggcaagggctgggaaagaaggcggccag cgggcagtgcgagtgtctacggcttggtagatgcagcactggcacctgctgatgtatgcggggatgggggcgggggaaggttctcat ctgtgtggggccaccactgtcctccacacatgggaaatgaagatctgaatgtgaccgaaacggcagtctctgaagggcctccttcctc ccttctctgccccagcttgctggctgttgggggcgggaaccggcgcacggccaacgtggtggcgcacgggtttaccaacctcttcatc ctggataagaaggacctgaatgagattttggtgcattatcctgagtctcagaagttactccggaagaaagccaggtatgtaacttttctttc gtttaataaaaaattgctaaggacaatgaatttctcttacataaccacagtacctttatctttttttttttttttttgagatgacatcttgctctgtca cccagtctggagtgcagtggagtgatcatgactcactacagtctcaacctcctaggttcaagtgatcctcctgcctcagcctcccatgta gctgggactataggcacgtaccaccattcccagctaatttttaaaatttttttctagagctggggtctcactatgttgcctaggctggtatca aactcctgggctcacatgatctgctctgacctcccaaagtgctggggttacaggtgtgagccactgcgcctggcctagtaccttcattaa aataaggaaatttcgtattgaaacaacactattatctcattcacaatccatcttcagattttgctaattctttaagagtatcctttgtaccttttctt cttttccattcctgaatcaggtctaggatcacatattgtaattaaggtgcagagtattcttagtcttccttgatctgggatagttcctcaggcttt tccaacctttagtgacttggacttcttttaagcatataggccagttattttatttatttatttatttatttatttatttatttatttatttatttattttgagac agagtcttgctctgtcgcccaggctggagtgccgtggcacaatcttggctcactgcaacctctgcctcctggattcaagcaattctcctg cctcagcctcctgagtagctgggattacaggtgcacaccaccacgcccggctaatttttgtatttttagtagagacggggtttcaccatgtt ggtcaggctggtctcaaactcctgacatcgtgatccacccccctcggcctcccaaagtgctgggattacaggcgtgagccattgcacc cagcccatacaggccagttattttacagaatgcccctcagttggggtttgtctgatgttttctcgtgattaccattagagtgtgcatttttggc aggaagaactcagaagtgagtcgcttctgcagcaaatgtatcgagaggcgcacgatgccatttactcccaatatcgatgatttttactttg attacttggttaaggtttctgtattgtaaacctattttcccttaaacctaattcctaagtaattagtgggtagtcaccttcagactatgaggatat cctgttccctgtcaaacttttacccactgcttttagcatcctttgatggttcctccctatgatagttacaaagagattgctatgatagttgtaaa atggaaattttgtaactccactccctccttgattattagctggcagtcagctgtgtggaagagctttctcttctctcccatttgtttattcatttct caatgtatataggcatggatttacttgattccatgaatactctgttaacaatcattattttgatgctcaaattagtccagtttggccagtgggca ctttttcaaggtggctcatttgtgaccttcgtgatatccctgtcatattttagcacttccttgcttactcaaggagcagtaagccacaaccaga tgccctggttcatcttgtacttcccttgcagtatcctggaaatggccatttctccaaggagccttgattccttttactgggggatggtttttaga aaccacgatctgagtcctaggagcagtcattgctactggagtgctactaggcctctgcagcagacagaaccaagaaacacacccacatacacaccaagatgtataagtacaaatatatatatatatataaaaaatacatatataatatattatgtatatataaaatacatacacacacatat acatgtatatgccaacttaaatttttctgtctattctactgattatctataaaacaatttcaatccagcaccacagaattcattctttttttttttgag acggagtttcactcttgtcacccaggctggagtgcagtggcgccatctcagctcactgcaacctctgctggaattacaggcatgtgcca ccacacctggctaatttttgtattttcagcagagatggggtttcaccatgttggctagactggtctcaaactcctgacctcaggtgatctgc ctgccttggcttcccagagttgctgggattacaggtgtgagccaccgtgccctgccaccacatattagaattcattctaatattctccctttt catacttgtaaatctcttctccaacactgagcaatacagggaagttcatttcagaatttccagcccatgtcactgtaaaaacaaaaagtaag gcttctaggtagagttcagtactggatgcagttcttttgtgtttagcccgagtatatatgatggaagtgttacgttcaaagttatctggagtcg ttctcttcctccactaccctttaatgtggttactttattcatttgaaatacaggtaggttcacttgtttcagtttacattccatttagggctcctgcc accctgatctttgttgactttttcttgttttcagtatgtagaacattaacatgttcctgaaggtcaacactctgcaaaatggggcactcagaaa agcgtcccaactctccctgtccccattgccccctgactgcaactcccattattacgtggctcgttgcatcaatttctgccttttctttcctctgc ttacttttgcgaaaatggacaagtccacagacattttctttttttcttttttgagatgaagtctcattctgtagcctgggctggagtgcagtggtt aggtcttggctcactgcaacctccacctcccgggttcaggtgattctcctgcctcagcctcctgagtagctgggattacaggcatctgcc accattcccagctaatttttgtatttttagtagagatggggctttgccacgttggccaggcttgtctcgaacttctcacctcaggtgatctgcc cgcttctgcctcccaaaattctgggattacaggtgtgagccaccacacccagcccaaacattttcttatttcttcttctttcttacacaaaata tggcatgctgtagattcttgtactttgctttgtttccacatgacgacaaaacccggaaatcattgtctatcagtgcagagtcagcttcctcatt cctttccacagtggcagaacattcctgtaaatggacatttagggtatttccacaattttgcaattacaaacagtggtgcaacgaataaccgt gtgcatccgtattttgcattgttggggaacaggtcattctttttagaaaagccggtttttgttttcaaacccttaagatcaggtttcaaattgag cgacaggcaacgtagtgtgtcaggaaccccacaataggtctgggaggctggcggggctcagatcccttccatgtgaggtgctggctg tgtgactttgaggaagttgtttaacttttctggtctcagttgtctcctatgtaaaataaaggtcataacccctactttgctgctgtaaattttttttg tttttttgagatggagtcttgctttgtcgccaaggctggagtgcagtggcgcaatcctggctcactgtaacctctgcctcctgggttcaaac aattcgattctcctgcctcagccaccccaatagctgggactactgcaggcacatgccaccacacccggctaatttttgtattttaagtaga gacggggtttcaccatgttgaccaggctggtctgggacttctgacctcaggcgatctgcccgccttggcctcccaaagtgctgggatta caggtgtgagccactgtgcctggcctgctgctgtgaaaattaaatagtgtgcaggaatgtctccggtggggtgcctgtcatacaagaga gctcagtgcatgttaataaggatcacttcaggccgggcgcaatgctgcatgattgtaatcccagcactttaggaggctgaggcgggag gatcatttcagcccaaatatttaaggttacagtgagttatgatcgtgccactgcactccagcctgggtgacagagcaagaccctgtctctc agataaataagcaattgttcctctgtagccatcaaattcaccctataatttttcagttgggtaaaacctttttaatccacatatctgggttttcct ccttgtcttgtatcacacattcttattttccccttcattctatacaatgacaccataaataaactgcacagagagctgtcataatagagggaa gaaactcaccagcatccattcctacagcacagtgatgtatcgagtacacagtgctgagttctggggacacaggagggaccaaacaaa actcacattctagtcagggaaccggcaatggaaagatgagcgatgatgtgatgcgtctggctacgaggaaacacaaagccacggga ggagcgggaagtgacggagagctgtctgagataagctggttggggaagctgctctgagcaggtggcatttcaacgagacctgcaca aagcaaaggagtgagctgggggactgatggctgggtacagcattctgggaagagggaacagcatgtgcagaggccctggggagg agcaggcttagtgtgtgggtggcacaacagcaagaaaattagcatagcttggtatgggatcagaggagtctaggggaagagattggg tcagacctcagaggaagatgggagccatggagagttttacacagaggagtgacttgaagtgccttaggttttcacaggagccctttggc agcttcgaggtaaaaggatgtagtgttgatgtccaagcatcatgcacacaagacactgaggtcccctggggaaggaagctgccccca aactcaccttacctggttgctctcaagggacccactggacccttccaaaacccagtctatctgaatttctcccagaaaatctccccacgtt ccccaccacatcagtggtgcgtgccagtgcctcctctccccagggacagggaggtagagtggacccttgtgattcatggactccatatt tgcaaattcgcctcctcactgaaatggacttgcaacgccaagatcaatactaacaggcttttgtggtcattcaaggacatgcccagagca gcaaaatacttgagtcaccccaagtgcacgctccccgctgaagtccctgttctgtcttgtggttttagctcttgtactgcgaagaggtgtct ttttatggtccatttaatgctactcttttttttttttttttttgcatttttgtgctttttgttggcgattgcactgtttaaaatggccccccagcatagtgc tgaagtgccgtctcatgttcctaagcacaagaagtctgcaatgtgccttacacagaaaatatgtgttagataagcttcattcaggcatgaat tacagtgctgttggctgtgcgttcaatgttaataaaacaataatctatatttaataagatgtctttagccggcacagtggctcatgcctgtaat cccagcactttgggaggctgaggtgggtggatcacctgaggtcaggagttcaggatcagcctggccaacatggtgaaaccccatctc tactaaaaatacaaaaattagccaggcgtggtcatgggcacctatcatcccagctacccgggaggctgaggcaggagaatggcttga ccctgggaggcagaggttgcagtgagccaagatcacaccactgtaatgcagcctgggcaacaaagtgagactctgtctcaaaataat aataataataatgtctttaaaaagaaacacacataagacaagtttattattattaattttagagacagggtcgtgctctttcacccaggctgg agtgcagtgacacgatcatagctcactgcagcctcgaactcctgggctcaagaaatctcccgccttggcctcccagtgttctgggactg tgtgtgaaccaccacgcctggccaagttcatgtattatggttgatgaatgttgtgatgggagatttgtaggaatctagccttgtatttcccct aggaacaatgtttccatttttgctaattcagtgtccgcagagatcttataggacataattactacaaatcatgagattcaactgcttatgactg aaaactggtaccaggtctccccaccctctcttcttgcccccacccccaccaggaggagcaaagctcaggaaagccatgcttttagctgt tacccatcctgctgggtgggttaggcaagagtttggcaaatcagatgggaccccaaaggcacctcccaagagctgtgcagaggcccccattcctgacagtgcctgtgcacctgtgtccacactgagaaggctgaggtgtgggtgggaggcagccaggcgaaggaatggggcta acaggcctcacagcaacatgggggggcgtgggaatgggggtctcagccatgagtcatcagagtggttgtttgctggccaggccagc ttgaccagagaggcaggggattgtgtatgagctctgctcatcaggggagatacactgtctgagaaccatagtcccggggacatctgag ttggtgagggccacgtgtgggattctgtccaagctccctgctttgcagatgggggacctgggtccccaaagaagtgaagtgctgagat ctccggcttgaccactccaaaaaagaggatgaagaggtcttatgtgggtcacttgctccaggctgattgctcaatctcagttacagggaa gcatacatcatcagaaaatgaggacaaatttccctaatctggaaatgccccagaagctagctcaatgtttccaaacagagacagctagt agcaatcagcaaaggtcctatacatagttattttgttagaaagaaaaagatagttcattttcacagcaccctcagccttcctgtgggctgtc acatcatgaattatggactcaaataacatgggggacagtgccgagctggcgcgtgtcagaggcgtctggtgaattatcatgtttgtgttct cctgggggatctccagctagaaatgcaaatcttgggcaattaatatcaacatgcacaatttaaacccattgcctctgattagataagcagc cagtaatggcctcatttttaaacaaacaagttttgctggagccttgatgaatgaatttgccaattcattgtctaggcccgctggttgcaggc agaattcatttgttcttgagacaaactgctcaccgcatgtttgtagtgtgaaaaattgccggtaaaaagctagggctctctttgatggaaaa tctacccttatctagagaaacagggtggtacatgccattagatgtgggtggcaattctgcttctgccacttcctggtaaccttgggcacgt cactccccctgctctgagcctcaatttccccaactgtcagaggcacaataatctcagccctttccatgtcacaggggtatttgtcagtgcc caggaagataatgatgcaatgtcgttttgtaatcattaatatctggggcactggggtcagaagtgtgggtttgaattctagctttgccgcttc ttagctgtgtggccttgggaaagttactcaccccctctgaacttctataccctcatttggaaatgaggatgaataataagacctacttatgg ggattgttgtgagacttaaatgagagagtatgtattagcataacagtattaccacaaatttagctgtttaaagtcacacatttgcgatctcag tttctgtgggttagaagtccggacatggcttagctaggtcctctgcaagattgtcatcaagatgtcttccagagcaggctctcatctcaagt ctggatgggggaaggacccacttccaggatcacatggttgttggtgtaattcagttccttggggttgtaggactggggccccagctccc actagcaattggccagtggctggcctcaggtccttgccatgtgggccttctccaacacagctggctgctccctacatcaaagtcatcaa gggaggccaggcactgtggctcgtgcctgtaatcccagcactttgcgaggccatggcaggaggattgctcgagcccaggagttcaa gaccagcctgagcaacatagggagaccccgtctctattacttaaaataaaaatcaaataaaaaataaataaagtcagcaagggagagt ccccttgcaagacagagatcacaatcctgtgtgacatattctaccagtgatagcccttgttgtattcgattgtttaaaagcaagacacgtac cccacccatcctcaagggaaggggaccacacagaggcgtgaacaccaggaagggggtaataggagccactgaagggtgcctgca aaggatggagcagagggcctggcatatacagacactcagtaaatgctgataattgtgatggtgacgagggggaggtgacagtgaag cagaaggagaaggaaagagctttggttccttttccacctgggcatcttattcccaattccctatgtgaaagtgtccttcaggatttcctaact cttctgctggttaaaaaaaagaaagaggccaggcgcggtgcctcacacctgtaatttcaacactttgggaggccaaggcgggtagatc gcttgagcccaggagtttgagaccagcctgggcaacatggcgaaaccctgtctctacaaaaagataaaaaattgtggtggtgcatacc tgtagttccagctactcaggtgggaggattgcttgagccatggaggtcaaggctgcagtaagccatgatcacgccactacactcctgcc tgggcgacagagccagaccctgtgagaaagagagagagagagagacagagacagagacagagagagagaggaagagaagag gagagaggagagaagacagaagagagaggagagagagagagacagaaaggaaggaaagagaagaatttcgtagctgtgtgacc tcagacaagttattcaacctctctgagcatcattttcctcatctctaaggtacaacagacagtaagacccacctcattacagttgttctgatc cttcaatgagatgcaatgcataaagtacttaacacgacgcttggcccagaaaaaactctctacaatcaccagccaaaatcaccaccact acacagctccagcttaaaggaggctgcagaagcgagtctttcttttcccccagagtgcgtctgtgatgtcaatggatgctgctttcttcca ggcgcatgctgagaagcaacaataagcccaaggaggagaagagcgtgctgatccttccaccccgggcgggcaccccaaagctctt caacgctgccctcgctatgacaggaaagatgggtggcaagggggcaaaaggcggcaaacttgctcacctccgggcccggctcaaa gaactggccgcgctggaggcggctgcaaagcagcaagagttggtggaacaggtaatgtggttgggaacccagccaccacgttcatt tcaatatgtgtcagggagaaggccaaggggtctccaagagtgggccacacatttagttttagggtcttccccctcctttttctatcagtca gggctcatttggttaccagtgacagagtgcaagctcaaattagcttgagaaaaacgggaattaggggtcacgtaatcgaaaagttcag gcaagcctggatccaaaagcccgaaagatgccaacaggacgagttttcccatcgtgtcatctctgctttattcagtgatggttttatctcca gacaggttctcaagagttctatctaggatgacatcctcctggcatagcaactccaacagaaagaaagcacctctttcccagggattctag caaaagttccaggactcactgtcattggacagaattgggtcacatgccaattcttgagccattcggtgtggccagggcactatggtgctc taaagaggcctgggtcacatacccgcccctggactgagagtaggctgggagcaggatctctaaaaggcagtcttgatactgttgcaga ggttagaggagagggtatgagaagcaaatgtccggccccagaagacagtgtgtcctcatatttaatactgtgtctgtgacaatttaagat atttgggtcaggcatggtagctcacacttgtaatcccagcactttgaaaggccaaggcaggaggatctcttcaggccaggatttcaaga ccagcctgggcaacatagcgagaacttgtctctacaaattttttctttttttctttttaattagccaggcttagtggcacatgcctgtggtccta gctacttgggaggctgagttgggagcatcacttgagcccaggaagtcgagactgcagtgagctatgattgcaccattgcactccagcc tgggtgacagagtgagaccctgtctcaaaaaatgaaaaataaaaatttgaaagtgaaaacctctcacacaggcaaaatgccaaatgtc cccacccccataaacctatgggggacattaaaattagttctgggacttttttctgtgaccaaagccaagaataccaagtggaatgattaca ttaattatgtctgttcacggctgtcctttaggaagtctgctggggatctggagagggactggggttaggcagactgggcttaacagtctac acagcagagttcaaaggagcccagtgaactcagtctcaatccagcctggtgtcagagtccaggagacccatccagatgttgtttgacagtagaagtctggtgagtgacatcaaagccatttggggtaccaagacattctccagatacagccaccaatggccccagactgaggaaa gatgtgattgcctggctcacaaactgtcaaggtcaaggtggagctggcaagtgggggtcttcagtgctggctgctgtgagggctgact agacaggaactccaagggtgctcccagtacaccaagagcttcagctggggtgggcttgtggctcctaagtggtagctcttcagcaag ggggctttaagatgggaattaaatttgcatgagtgttccattaaaaggcacttcccttccaaatcagaacaaccagggcagaatggcag agtacagggagctcttaactagacattcagagacacaaatccaaatcccagctctgtcacccactagctccatgacccaggaagcccc tcaagctttccgagacttggattactgagggttgcctcatgaggtcataaagatcaggcaggattaccttggaggccttttagaaaagcat gttgaacacagcaaagcacttttctaacttcccatgagctgctgattccagccatagcaccatgcttcaggaagttccacgatgctcacc atctaagcatcctgccctggaaatgtgttgtcaagagtttagggtgggctgggcacaggtggctcacccctgtcatcccagcactttgg gaggccaaggtgggcagatcacttgaggtcaggagtttaagaccagcctaaccaacatgtaaaaccccatctctactaaagatacaaa accttagccaggcatgatggcacacacctgtaatcccagctactcgggaggctgaggcagaagaatcgcttgaacctgggaggcag aggttgcagtgagctgagatcacaccactgcactccatcccgggtggcagagcaagactccgtctaagaaagaaagaaagaaagaa agaaagaaagaaagagagagagagagagagagaggaaggaaggaaagaaggaaggaaggaaggaaaaaaggaaggaagga aggaaagaaggaaagaaagaaagaagacaaaaagaatagagtgtagggtggacaggggagggtgggacaggggcaccaaaac atagggaaacactgagacagagggagaagcatgtcccaagagaggccttgggttcttggagcattttggctgccccctctccattcctc ctttccttcaatatcctatgacttgagcaaatggcgcaggtctctctgcacaagcacattgactgctctcacttgaaactcacaaacaccac ttcacatgggtcttgggcagctgaggaccatcctggcctcctctgggggctgctccctctccagtttccttccgtgcagagatggggtca ggcagagtgggaggggccacaggtatccgcaaaggcttgatttaattacccagtcccctactgatgtgaagaccatggccatcgtttgt gggaggcagcacaggctggaggctggaggtatccatcaggtccccacacatggggtcagcgcagcccaatagcaggggtaccca gccccagggtatccctgaggatctcaacctttgagacttcccagaaagggggcccagccccagcagcctctcccttcatcactttttac cctgtttgcttggagtttttctcatccactccaggagtgcccacccctggtaacagatggttggttcagcctttgcagaggccttgcgggta ctccctcagagtcactgcctggcttgcttcaatagctcgggggaggggccctgaacttgaaacaacagcctcatcccagcttccagcc ccacagcctagatctaagacatcccaagactttccacgaaatccaaccgggacttttaatctgtacacttccccataatgtattttgtttctc ctaaataaccccgatgcggctccaccccacccactagatttcacccgctgtgtgttccgctgccctctttgggggtccaaaaaggctgg aacagggaggtgtccaagacccttccaagacccctccgagagtctgtgcctcacgagtcaccctttctggattccaggccaagagctc gcaagacgtcaagggagaggaaggctccgccgccccagaccagcacacgcacccaaaggaggccgccaccgacccacccgcg ccccggacgccccccgagcccccggggtctccaccgagctctccaccgcctgcctcccttgggaggccggagggagaggaggag gggccggccgagcccgaagagcactcggtgaggatctgcatgagcccgggcccggagccgggagagcagatcctgtcggtgaa gatgccggaggaaagggaggagaaggcggagtaaggtggggtgaggcggatcccgcgcgcagttccagcaggtgtgtccccag cgcccgctgcgcccctgcgccccagcgccccaccttcccccacggctcaagagaagatgcttttccgtagtcgtgacctcagtggct gcagctctgaccgtcccgccagcacgccagccccgactcagctcctcgcggggctgggcctgagctcgacaagttgcatcaagtgtt cgagtccctgagctctcactatcatttgagagccctaccttttcctgactgttcctctttttaagaacaaaatgattttccacttttaaagttcat gggtgagggataaaatgaggctccaatacatgggaagctttgctgaaaaagtaaagtgttattgaatgtgtggggtttcccctcaggaat ttgtctaacacatttcaaggatagaaaatacttcactgccgggcatggtggctcatgcctataatcccagtgctttgggaagccgaagca ggaggatcactggaggccaagagttggagagcagactgggcaacatagcgagacctcatctcaaacaaaaacaaaaacaaacaaa aaaacaatagccaggcgtgatggtgtgcacctgtagtcccagctactggagaggttgaggcaggaggatggcttgagcccaggagtt tctggctgcagtgagctggacacagtgagaccctgtctcaaaaaaactaaaagatagccctgcacagtggttcacatctgcaatccca gcagtttgggaagctgagatgggaggattgtttgagaccaggagttcaagaccagcctgggcaacatattgagaccccatctctacaa aacagcagcagaaacaaaagcaaacaacaacaacgtacacaaacaaaaaaacaaagaaaggagggcagcccgagctgttcagct gagaattgccatggttctgagctgagcttgggacgcggttttcctttgcgggcaatcacagtagggggctagatgcccattaaggtagtt agtgcttatcttccccagactctattcaattccaatgcaatgccattatctcagaacagatagaagggaaatggacttactggccaggagc tgctagcgtttctgcctctgccaggaggaacagcccggtaggagcatcctctgcctgtcatcctgagaaaccccattcgccagcaggg atcagtccgttactcacttttgacactctcgctgtagagatggtggcagcaacttcaattaatcctagaaaatgaaatctcccagctgatga cagcacttacattccctaaagctttccagatcgtgtcatgttgcttttatgattcagacaaaaaaaaaaaaagaaaaaagaaaaaaaaaga aaaaagggtcattcagggaatgaatatttatcaagaggattttatttgcataccaaatggctcttgggggagtgagtgattaaaaacaaaa tctagtattctagtattaggccatttgtttccacctgcaacagtgaacataaaggacctggaattccaataggtcctgcccattttatataaaa ttacaggaaaatgtgagctgctcagaggcctttgaaggtttgcaaatgtctgtatgtatttgtccatccactaaaaacaacattcatttccac tgtggaaaaatgaaagaatgtgggagtccccaggctggtctaagcatttccaaaccccagagtcagaaacagatttgtctagttgatgg cttgtcacgtgatccgggtaatcatcacattgaatatagtgatgattgaccttattattttaccctattttttttttaaaaaaaaaagctttaaaaa tgtattttaataggcaataacatatttacatgggtacaaaaggacatccaataaaaattcttcatcctaccccagtacccagcttccttggttc cctctctggaagcaaccactattaacagtttctcctgtatcatttcagagttattttatgcttatacaaggaaatatgtaactgcttcttcccttccttttaaattttaacagaagtggtaacacatattacagccactgttatttttcacttaatttactttggagattgttccctgtccttaaagagcatg cttattttttcatgacggcaatctagtttgttgtatggatgtatgataacatttaaacgactctcccattgatgggcatttgcgttgtttccagtct tacgctgttaaaaataatgctgcaataaaacaatcttgtgcagacatcatttcactggtaaacattttttaaatgggggaactctgcgagttt aggcagttatttttccttcgtctccatttactgtttgtatatttgtaccacatagtctaagatcaaagcatggagaatgcttccaccattagcaa gtatttcagatatgctaataaacacattgcttttccgtaaatggcaaaatgcttcaacagcaccttatctgtgagatcctattttataaggaat cttttttatacgaatgtacttattcttttagatatcagttcttgaatgtatactccttatgatatacagcaagagattcttgaggttcccaagaatt gtcctggaaagaaaattgattttgaattgagattagcagtgttttaatttcgctgagataactcctttaactcagcactttcacaaaacttaaat attaaataaattgaatttgaggctggcattcttctctaaattgaattggactggttgtttgctgaggtatgaatggagcagctcggaagtatta taattccgtagctgaaacaggacgcttatttatgtatgttcaatctctactcatttaaaatgtctcagcatttcctgcatttagatagaaaataa atagtcaatttttttttttaactacataaatattttaaatagaatgaaagccaagaagctgtttcaaattttattatcaaagtgtgttcagctacct ggaatatcttagtctgttttcttagtaaacatgaacactttatttacactctgcctttcatattacaatgcatttaattaaagtatttagacaaatgt tcctggcttgtggattcatttcagaaatcagccatgggtgacagacacctctgtgagaaaaattaagtaaatgtcctggcccaggcactct agcaaatgtcctacggacatccttgtggctctaatccatcctgtattcctattgctggaatgatattcttatcaaacctctatggccactttgg gaagctgaggtgggagaattgcttgagcccaggagttcaagaccagcctgggcaacatagtaagactccatctctattaaaagaaaaa atagtctgggtgcagtggctcatgcctgaaatcccagcactttgggaggctgaggcaggcagatcacctgaggtcaggagttcgaga ccagactggccaacatggtgaaaccccgtctctactaaaaatacaaaaaaattagccgggtgtggtggcgggtgtctgtaatcccagct acttgggaggctgaggcaggagaatcactgaaacctgggaggcagaggctgcggtgagccgagatcatgccactgcactccagcc tgggcgacagagcaagactccgtctcaaaaaaaaagaaaaaagaaaaagaaaagaaaaaatagggccaggtgtggtggctcacgc ctgtaatcccagcactttgggaagccaagatgagtggatcacctgaggtcaggagttcaagaccatcctggccaacatggtgaaaccc ctctctctactaaaaatacaaaaaaatt

[0267] SEQ IDN0 14 aagccaacaagaatgccgtggagtctgtgcagtatggcgtggccggcagcgcagccttccttgagtgccagccccgctcgccccaa gccactgttaagtggctgttccagcgagatcctggtgaccggcgccgagaggtgagttcctgcgcccggtgctgcaccgtggatgtg agtccttgcacagtggtgaaatgtagtagaggagtgatctggagcaactctcatgctgtgaaaccccttccaagctcccttccctacatg caaatcctttgagtgaaaacagttacatggaaactccctgggattggcctcattctatggaaagtgtttctgggtatgcctgttcacatgaa aattccatatgcataccacattgggggtgtaccttctacaaaaacatcatgtgagtgagctccagtatgtggaaattctatggggccctca ttttatgaaaatgccctacaggtgtccctgtttatgtggaaattctatgtggttatcccaattggatattctcgttaaatggaaattctggctgg acgtggtggctcacacctgtaaccccaacactttgggaagccaagacaggcaaattgcttgaggtcaggggttcgagaccagcctgg gcaacatggtgaaaccctctctctaccaaaaatacaaaaattagctgggcatagtcatgcatgtggtcccagctacttgggaggctaag gtgggaggatggcttgagcctgggaggcagaggttgcagtaaaccgagatcattcaactgcattctagcctaggcgacagagccag accttgtctcaaaaaaaaaaaaaaaaaaaaaaaaggaattctgtgtgtgcttcaaattttcctcattgtacagaacctccttatggtcatcac tcctgaaatggaaatctgtacagatgccttattcaaatatctctcattatgtagaaacttcatgtggtttttctggcatatggaaatcccacatg ggtgcctctacacggaatggccacaaaggtggtctgatctggttgctggctagggccatagggtggtcacagagcctctgaaccccct ctccttgcccctgcagattcgtgcagaggaccgcttcctgcgcacagagcagggcttgttgctccgtgcactgcagctcagcgatcgt ggcctctactcctgcacagccactgagaacaactttaagcacgtcgtcacacgagtgcagctgcatgtactgggccgggacgccgtc catgctgccctcttcccaccactgtccatgagcgccccgccacccccaggcgcaggccccccaacgcctccttaccaggagttagcc cagctgctggcccagccagaagtgggcctcatccaccagtactgccagggttactggcgccatgtgccccccagccccagggagg ctccaggggcaccccggtctcctgagccccaggaccagaaaaagccccggaaccgccggcaccaccctccggacacatgaggcc agctgcctgtgcctgccatgggccagcctagcccttgtcccttttaatataaaagatatatatatatatatatatatataaaatatctatattct atacacaccctgcccctgcaaagacagtatttattggtgggttgaatatagcctgcctcagtggcagcatcctccaaaacttagacccat gctggtcagagacggcagaaaacagagcctgcctaaccaggcccagccagttggtggggccaggccaggaccacacagtcccca gactcagctggaagtctacctgctggacagcctccgccaagatctacaggacaaagggagggagcaagccctactcggatggggc acggactgtccaccttttctgatgtgtgttgtcagcctgtgctgtggcatagacatggatgcgaggaccactttggagactggggtggcc tcaagagcacacagagaagggaagaaggggccatcacaggatgccagcccctgcctgggttgggggcactcagccacgaccagc cccttcctgggtatttattctctatttattggggataggagaagaggcatcctgcctgggtgggacagcctcttcagccccttctcccctcc ccgcctggccagggcagggccaccccactctacctccttagctttccctgtgccactttgactcagaggctgggagcatagcagagg ggccaggcccaggcagagctgacgggaggccccagctctgaggggagggggtccgtggtagaggcctggggccggtagaggct ccccagggctcccttatgtccaccacttcaggggatgggtgtggatgtaattagctctggggggcagttgggtagatgggtgggggctcctggtggccttctgctgcccaggccacagccgcctttgggttccatcttgctaataaacactggctctgggactagactttggcttctctc cttggtcaggggatttagagccacacgtggtgggcctggggagccaggtggggggctgggaagggagtagattggctccatttaaa gaaacccaacatgtagaaaccacatttagtttgacaaacctgggctgctgcaccaccccaaacacttgggtagtgcgaagggctgtag agacccctcagggcccactcactcctcagggtcagggagggtgggtggagcctgaaggctgaaagtggcagaaatgcaacggcct cactcctggacagatccccatcatccaacagggtgggtgtagaagctgccagcctgtggtaggagccagcattcacttcccacgcata ctatttagagacagccgttagtgctcagtgtctggtgccccggcacccccttgtggtctcttgagtctgtggcagagttgagtgctgaaa ggcagccaggttccagggaagagtccagccaggaggctcagggtgcctggaggagcctttgagagcccaagatttgggttttcctat ggaagttgagggtgggaagtctctagaacctgggagctcaacctgaacaatttctgcccggcagtgagccctgcacctgaacagagc accagaccccctaagctgaggcaggcaggccctggagataatcttaggcaggcctcagagcacccacagcagacagagcaggag caggataggtgagccccattcctccataccccagggaacactacacctgctctgggatgcaatttctccacctggcaacaggcctgca aaacattctctggaagagcctggccacttctgcagattagagactttgttgcacccaaattcaagtgtacatataggcacaggcgcctgc aagcatgcatgcacacacacagtcatacacaggcagagatggacatgcacaggcacacatatgctcaggtacaaacaggcatgttta cacccctgcgtataagcaagtgcacacatgcaaacctacacaaactgcaaaacacacaagcctatgtgtgcatgaagagaatacaca aagcatattcacacatgcatacacatgtgtatacatgtgcacacatacataagcatgcgcacaggcacgtgattcctataagagcacatg cactcacaaaggcataaacatgcaggcacacatgtacacacaggtattcatgcacaaatgtgtcaagttattgctcgcttggtgccagc aaaagacacacccatggccgggcgaggtggctcacacctgaatcccagcactttgggaggccgaggtgggtggatcacaaggtca ggagatcgagaccatcctgaccaacatggtgaaaccccgtctctactaaaaatacacaaaaaattagctgggtatggtggcacatgcc cgtaatcccagctacttgggaggctgaggtgggagaagcttaaaccagggagttgaaggttgcagtgagccaagatcgtgtcacagc actcaagcctggcaacagagtgaaactctgtctcaaaaaaaaaaaaaaaaaaaaaaaaaaaaggccaggcacggtggctcatgcct gtaatcccagcactttgggaggccgaggcgggcagatcacgaggtcaggagactgagaccatcctggctaacacggtgaaaccctg tctttactaaaatacaaaaaaagtagcccgacgtagtggcgggcacctgttgtcccagctactcaggaggctgaggcaggagaatgg cgtgaacctgggaggtggagcttgcagtgagctgagattgcgccactgcactccagcctgagcaacagagcgagactccatctcag aaaaaaaaaaaaaaaagacacatacaccgaggcacacagagcagatatgcatgcccaccacagtccgctggaagcagaggacctc cttggggcagctccagcctgtgatatgggatgaatgcaatgcccactgtttccctctctctggattccctgcaggtcataaaatcccagtc cagagtcaccagcccttcttaaccacttcctactgtgtgaccctttcagcctttacttcctcatcagtaaaatgaggctgatgatatgggcat ccatactccagggccagtgtgagcttacaacaagataaggagtggtgctgagcctggtgccgggcaggcagcaggcatgtttctccc aattatgccctctcactgccagccccacctccattgtcctcacccccagggctcaaggttctgccttcccctttctcagccctgaccctact gaacatgtctccccactcccaggcagtgccagggcctctcctggagggttgcggggacagaaggacagccggagtgcagagtcag cggttgagggattggggctatgccagcccgattagaagggttgggggggctgagctggattcacctgtccttgtctctgattggctcttg gacacccctagcccccaaatcccactaagcagccccaccagggattgcacaggtccgtagagagccagttgattgcaggtcctcctg gggccagaagggtgcctgggaggccaggttctggggatcccctccatccagaagaaccacctgctcactctgtcccttcgcctgctg ctgggaccatgggggctggggccagtgctgaggagaagcactccagggagctggaaaagaagctgaaagaggacgctgagaag gatgctcgaaccgtgaagctgctgcttctgggtaggggtgtgggcccaggtggggccactgccaccaggtcattggtctggaggcag gcaggcagcccttctgtgaagcaaggattccctgggaagcagtatgagcctgtgacagagtagggcacagggtgggagccctggc caagcaaggccaggcctgacaggtggtgtcctagtcccccagggctggacatcccacaaagcccctctccagacggctctgcctca ggtgacccaccctccccagaagtgcgtgccaaccacactcttgcacgggtgtgtgatgccgacttttccgtggctgggttcccgcaag cttcatgcagcctttggtttgatacgcctgctaggcaatgctcagaagccaggagcctcactgggagcccgttattgggctgctactttg ggaaggggatggccatagtcctcctgggagtggggagcctctaggaccacccagatccacaacgaatggccctcaggtgccagag ggctgtggctgatagacctacaaagcagccagaggggctggcgggtactgaggcccagtgggagtagataaagccgcgtccactg gaccatgggcactgaggagagatggggctgatccaggcctgcctcaaggtcactgcctgacctctcaaggggaaaaagcagcttca agaggggctccacccaagctccccagggtctggtcaaccagctggaggtgggtgagggaccaagccagtcccactttcccccaag gcagagaggctaagccggaaatctcattaactgtgcccaacgccctcgcaggggcaagctcagaaaggatgtcaactctgccctccc tcctacccaaaagagctaattccctctgagcctcctggagacacctgcaccccttcaggtgaaatgcaaggggtttacccgcctcaaat aacccagtgcttgcggtaccagaggctcccacgaggtggcgatgacgctccgggatttcttcattgcagtctcacagcataataaagg ctgcctgtgccggggtcttggggactcccctgtcccagaacagttgaggtcgttgagtcccctcgttggcttcctttcttcactttcctggc atttgcaaaccttgtgacttcaggatgtcaggacttaatttggatgggggggtaggtgtggctacggggtcggggctggcgctcaggc ctcttcgctgcgggtccaccctgccaactcgggaggtccccgtcctcctggcctccttgctggagggggcaggctggtcagcgcagc tctgaggcgccgcgtctctttcaggtgccggtgagtccgggaagagcaccatcgtcaagcagatgaagtgagtgcccctgcctgtcc cgaggcccctgggtctgggtccttccccacttccccacgaggcgctgattctgctctcctcggcctcaggattatccaccaggacgggt actcgctggaagagtgcctcgagtttatcgccatcatctacggcaacacgttgcagtccatcctggccatcgtacgcgccatgaccacactcaacatccagtacggagactctgcacgccaggtgtgccaggaggcgggctgagcgggcctctggggactcgaggctcgcggct gggcccgagtccctggccgccaccagccactctcaccctgcccccaggacgacgcccggaagctgatgcacatggcagacactat cgaggagggcacgatgcccaaggagatgtcggacatcatccagcggctgtggaaggactccggtatccaggcctgttttgagcgcg cctcggagtaccagctcaacgactcggcgggctagtgagcgcgcgggcagcgcggggcgcggggcgcggggcgcagggggc cctccacgcctccccacccacctacggccgggtctcgcgcagctacctctccgacctggagcgcctggtaaccccgggctacgtgc ccaccgagcaggacgtgctgcgctcgcgagtcaagaccactggcatcatcgagacgcagttctccttcaaggatctcaacttccggta cgacccatacgctagcccaggaggtcactgccccaggccccgtcctgccccggggaccccatccctgcgatagaccctgccccttc cctgataccccgccagcagaaagggtggtagtcccggccgagagctccccgaccagcacagggcgaagggatgttgcctgtggg gcccggggcgcaggttcaggcccccgcggccccgcaggatgttcgatgtgggcgggcagcgctcggagcgcaagaagtggatcc actgcttcgagggcgtgacctgcatcatcttcatcgcggcgctgagcgcctacgacatggtgctagtggaggacgacgaagtggtgc gtgccaggcagggcctgtgttccaggggggcgaggaggagctgctggtccctggaggcggagaccgcgcgctgggctggggga gggcacgggaggggatgcctgtcccgggcggcctgaggaggcccggaggcgttcagcaggcccatctggggcagtgcgggga gccctctgagagccagctgagcgggaagccccgcgtgcccgggagcccagagagcaggtgctgcgggtcggacgctaccccgg gtgcccaacagctgctgccctcctcagaaccgcatgcacgagagcctgcacctgttcaacagcatctgcaaccaccgctacttcgcca cgacgtccatcgtgctcttccttaacaagaaggacgtcttcttcgagaagatcaagaaggcgcacctcagcatctgtttcccggactacg atggtgagaagtccgcaaggccgccaggcggcgcccccgccccacgatcgcggcgcgcaccccccgcacggggaaggaaggg ctgagcagagtgagagctcccgcccccgcaggacccaacacctacgaggacgccggcaactacatcaaggtgcagttcctcgagc tcaacatgcggcgcgacgtgaaggagatctattcccacatgacgtgcgccaccgacacgcagaacgtcaaatttgtcttcgacgctgt caccgacatcatcatcaaggagaacctcaaagactgtggcctcttctgaggtaggtcgctgccctctccaggctcttgcctcaataccc cagccccgtccagctccccaccccacaccacggccctggagcctcacccccaaatcctggagcctactgcctccaggccccacccc agctcctcaggctccaccccaccgacagaggccccgccccaccccacaagtcccacccattgaatttcttttggccccgccccaaga cctcaggtgtcacagcttccagggccgccccaccacacacaatgcctaaacccatttattttgggtctcagggatgctgactgggtttcc ttttcacaggtgcctgaattcatgcgtgtcagtcaccctgagacctggtagccctagctgccttgcagccccaaaccccaggaccctatc agccccctgggactccctggccccaacctgccacctcaccaaagctctgagccctgctagccttgaggcgcggaccctcccccatac ctcccacagtatcccagcgcctcatgcccccaaccccaagccctgcccttcaccgcccggctgcactggcctctaggactcacagca gccagccccagctaggaacgggcaggacttggggcagctgaagctcacggtgactcagcagtgcccaactgaccaatctcccgca gctctcctgcccctgggggcccacgactcacctggtgggcctgggctcagcaaacagccctccctcccagtatttcatcaaggacag ggaatatccctcccacacacaggccagctgctcaccctgccacctccatgtgacaagtttgacacctgtctacccagagccagtgact gtccctcccagcagcctccaaagggtccaaagtccaagctagctctggcctgtgtggcccccacataaataaattagtggggagggg gcagacctgggggcccttgggtgccagagtgagacacagcaggacccttccccaaatactaggctcagagcaacctccagccttctc cagcaggatctgctccctgtgtccttggacagcagatccacacaccacttccccacccacacacagtaccctgggctggttggacaac aggatggtgtgttaaggggttaagggagggtactgttgacaccagctatggccaatagctgggctgtcctgaggggtggctggggac agagtccagctcccttggatcagcccagaggattcaggataggacaaaggccctggctgtgcccaaaagggtctacctcaggtggg gttacaagcaaacctgggtgaccctcttgtccctaccagcagcttctcttgccccttccattcagggttccctggcaggggaagctgaga gccatggctgaactatcagggacaaaggcccatgtccccacatccctgctccctccttcctcatccagcaccaaatctttgcttatgtcca caggccagggcctgtgctgcagtcggggacaaggagcttccgtctggcaaggccggggcacaatttgcactcccctcagctagacg cacagactcagcaataaacctttgcatcaggctccagctgtcctttcttggtggaggacttaattatcacaagtcatgggcatttattaagt gcccagtgctgggttgggcatgaagtgggaagatggcccctcccaggaagaagtacctggcctgacaaggtggggcactcttgggg gtatgggaccaactcatggcttttcacgggagttgaggagagaggagctgtggaaaatattcactgggacagtcttggatcaagaggg agttttgaggtggaggctcattctggcagggaccgtagtgtctaccagccccagaaacatgggcttatggccacaggagttcagtgga gcaagagcaggggaggagagacgtggacaggtgcccaaagccagtcggagggcctgggctttctcagaaggtgatggagagtctt ggaagccctcgaggcaggaacataattgcagggctgggattagggtgagggaagtgaggcacactcaccttgggtgcaacatttaa ggcgatgccaaaaaatttagtaaccaaggtaaataatattaggataatatttttaaaaatcaaatgaatgcaaaaccccacaatgaatgaa atatcaaaatccaacagaggatcaaacagaggcatgctaagatatattggggcttgaagcaaagggaaaactatttgttgctatatgtttg tagggattttttgccagttttaaaaatacatgtatcataaagtttactatctcagccacttgccggtgtatagtttggtggtgttaagtacattca taatgttgtacaaccaccgcaactgttcatctccagaactcctttcctcttgtaaaactgtaactctgtacccatgaaaaaataaccccccat tcctgccttcccccggctcctggcatccaccattctactttccatctctatgaatgtgactgctctaagtgcctcagatgtgtgggtccatga agtctttgtctttttgcaactggcttatttcacttagcatcatgtcttcaaggtttattcatgtgtagcatatggcagaatctccttcctttttaagg ttgaataatattccattgtatatattccacactttgtttatttattcatctattgatgaatggttacatctgccttttggctattgtgaataatgctgct atgaacatgggtgtacaaatctctctcccagcctctgttttttattattttgggtgtatacccagaaatggaactggcaaatcatatggtgaatcttttttttttttttttttttttgagacagagtctctctctgtcgccagcctggagtgcagcggcacaatttcggctccctgcaacctccgcctcc caggttcaagcaattatcctgcctcagtctcctgagtagctgggactacaggcacacgccaccatgcccaactaatttttgtgtttttagta gagacggggtttcaccatgttgtccaggatggtctcgatctatctcttgatctcaggtgatctgcccacctcggcctcccagagtgctgg gattacaggcgtgagccactgcaaccggcctcatatggtgaatctatttttattttatttatttatttatttatttattttgagacgaagttttgctct tgttgcccaggctggagtggcaatggcatgatctcggctcactgcaacctcctccttccatgttcaagtgattctgcctcagcctcccaag cagctaggattacaggcatgtgccaccacacctggctaattttgtatttttagtagagatggggtttcaccatgttggtcaggctggtctcg aactcccaacctcaggtgatccgcctgccttggtctcccaaagtgctgggattacaggcgtgagccactgcacctgacccctttttttga aatgaagtctcactcggtcgcccaggctggagtgcagtggcacgatctcagctcactgcaatctctgcctcccgggttcaagtgattctc ctgcctcagtctcccaagaagctgggagtataggcacccaccaccatgcctggctaatttttgtatttagtagagatggggtttcaccatg ttggccaggctggtcttgaattcctgacctcaagtgatctgcccacctcggcctcccgaagtgctgggattacaggtgtgaaccactgc gctcagcctacttttaatattttgaggaacagttttccacagcagctgtaccattttacattcccaccaacagtgcacaggtggtcccccttc ctctacatatttgtgaactcttgttattttctgggttttgttttgtttgagatggggtctcgctctgttgcccaggctggagtgcagtggcacaat ctcggctctgcaacctctgcctcctgggttcaagcaattctcctgtctcagcctcccgaatagctaggactacagctgcatgccagcatg cccggctaatttttttttttttttttttgagacggagtttcattcttgttgcccaggctggagtgcaatggtgtgatctcagctcactgcaacctc cacctcctgggttcaagccattcccctgcctcagcctcccaagtagctgggattacaggaatgcaccaccacatctggctaattttgtatt tttggtagagacggggtttctccatgttggtcaggctggtctcaaactcccgacctcaagtgatccgcccgcctcggtctcccaaagtgc tgggattacaggcatgagccaccgcacctgacctatttttcttttttaatagtagtatctcttggtagttttgatttgcattttcctactgcttagt gatgttgagaatcttttcctgtgcttattggccatctgtatatcttctttggagaaatgcctgtccaagtcctttgcccatttttgaattgagttatt ttaatgttgttgttgagtcttagaagttctctatatattctggagaataattccttatcagagataggatttgcaaatttttttcccactccgtggg ttgccttttaaactcattttatgtatttttaatggttaatttttttccagaacattaaatagttgaaaaaacttgtaaaagtaggtatgttaaaactc acattattttaagcttgaatattttatttataacttaaacagaattttttttttttgagatggag

[0268] SEQ IDN0 15 gcatggtgaaaaaaaggaataaaatactgatacatgctgcaacatgtatgaaccttgaaaacattatgctaagtgaaataaatcacacaa aaagctacatattgtataaatccatttatatgaatgtccagaataggcaaacccatagagagagaaactggattagagcttaccagggga tagtgggagaagtaaggttggactgattgctaataggtatgacgtttcttgtggggttgacgggaattttctggaattagtggtgatggctg tacaacacagcaaatacgatggcctccctcattcaaagcccacctggagtcacagaactgggagtattaaagataaattattcaactcct ttgttttgttgatagggaaatcaaagccttgagagaggatgggactttctcaaagttacaaaatgagtcgttggcattgctggacctaaaat ctcgtattcctctttctagtagtacatgcacttttctaaactagattacacaagacatctccccaaactctactggtagcccctattgccaata aaagaagcttcagattaccagtttcttccctgacgtctgaaatcaggtccatttcataagttagtgttaggcgttaggtgactcactccaaat accaacttatttgtacctacctagcaattagactgaagtttttcaacagccttgccttcctcattctggcatgttgaaaaatgagataaaagta gattttttggcaaactgttgcttctatttctcaccagaacttggtggttctggtggacatgctacggaaaaggacatgtgctgagctctttgg gaaatctctttttaaagacagggtctcattcgctcaagctggaatgcagtggcatgatcataactcactgcagcctaaaactcctggcttc aagtgatcctccagccttagcttcctgagtagttggaattacaggtatgagccatcatgcctggctctaggagacattttaaaatgggcca agttttacttctatagttcaaggagtttgtccttgatgaaagtacaggtgtgcagaaataacacaacctgaaatgtagaaactccccttaga atggaaggaaaccaatagtttgggagttgtgagtagaaaaagagtttagttggtacatcaaagaagaatcaacagaagtaatgacattc ggacagggtggtaagggacaggtggagggcacagcaagaatgaagaggcaaaaccgaaaaagaatgaaaacattagggtgaag cttggaatatgtgtagagaaattgtggaaaacaaaataagaagtgttggggacaaattctagagagctattaatgccaggattaagaaaa agtgtttaatttactaggcaacaggatgttgttgaggggtgggttaagggttagggacagacactgtcattcacactggtaaagccttgg agacaaaatttggcattatttatcaaactccttagacagctgcacactctttggacctggaatttcacttctaggtattttctactaagaaatat aaggatgttcattacaacattgtttctaatattaggaaattagaaacaacctaaatatctaactatgggataatggttctcaactggggcgat ttggaaatttctggagatatttttggttgttacaacttgaggtgggtgttattggtatctagcggtagaaattagagtaaacatcctgcaacgt acaggacagcctcccacaacaaagaattactggacccaaaatgctgatagtgtcaaggttgagaaacctggctgtggacggtagtact gtggtgttctggtacctccatacagtgaaatgctttgcaatcattaaaatgaatgggacaggctgggcatagtggctcatgcatgtaattc cagcactttgggaggctgaggcgtgtggaccacttcagtccaggagttcaagaccagcttgggcaacatagtgagacccctagctcta tgaaaaatacaaaaattagctgagcatggtagtgcacaccttcagtcccagctactcaggaggctgaggtgggaggatcgcttacgtc cccgaggtggaggttgcagtgaaccgagatcacgccactgcattctagcctggctgacagagtgagatcctgtcaaaaaaaaaaaaa aaaggaaatacctttgaaaggttttcatgatatagagttcatagccctgccaccctgaatgtgtctgatgtctcctgatcttggtagctaagc aggattgacctgttattattcgagaggagaccactagctggcctccaacttctgggctcaagtaatcctcccgcctcagcctcttgaatagtataggtatagcccaactcacttggatcttctaattcaattaaatcctgacattttctacctggagatagcctcagatcccataggttgagag ctcagtcccacaagactgttcccctgtgcctcgcctttgggcctgcagaacttctgaccagccagtttcaagttggggttcccatgacac cctctttgggttcaattaattactagagtggctcacagaactcaaggaaatttactaacctttactggcttattataaaggatatcacaaagg atacagatgaagagaggcatagggtgaggtgttagggaagggactctgggcttgcacggtccctgggcttgccaccttctaggaacc tccttgtgttcagctgtccagaagctcatctgagccctgtccttttggatttttatggaggcttcattacacaggcaagattgattaaaccatt ggccactgatgatcaactaaagcttcaggcccctttccccttcttggaggttggaggatgggctgaaagtcccatcctctaattgtggttt agtctttcaggtgaccagcccctcagcctgaagctacctagggattaccaggcactggtcaatcattggtatacaaaaatacatcactct ggagattctaaggattttagaagttgtgttacctggaaatggtgttgaagaccaaatacatgtatcacaatatcacaatactgaataccaag gatgtgtcaggcacccttgcaaggcctagggaatcaacactgacaggtccctgccctcatgaagctcacagtatagtgggaggccag acaataggacaaggaaacaaaaacaaacatgataattgctgatagtaataaatgtaatagaaaaaataaataggctgatatgacaggca gaaaccggggagagggtatttgggggctacttgggataagaaaactagagatgacctgtatgacctgaaatctggaagttaagaagg agatggcatgcagagagctgagaaagcgcattaggttagagagttccagttccttatgatgggagtgtctggaaggtttgtgctgctac acagcctgccagattggggggtgagcactcgtgcacactgctggatgagtgcagtgccccaagaatttattacactctacaataaagg aggcaatctgcttcccagtaaaggtcatttttgaagttcaaagtgctagcaactgtctataattatgcgttccacatagtttacaacatagag gtgccaggctatagtggatatagtcatggaagctaaagtaagtttcatccagaaagaaaagtgctctgtctccttcctagtgggcaggaa gggcattttaatgaagattctggtatttcaagattcactggagttgctagaatgtggaccattgtgtgtatttctttctttccttttctttttttcttttt gagacaaagtctaggtcttgtcgcccaggctggagtacaatggcgcaattttggcttactgcaactttgcctcctgggttcaagcgattct cctgcctcaggctcccgcgtagctaggatgacaggcagtgccactatgccctgctaatttttgtatttgtagtagagatggggtttcaccc tgttgcccaggctggtctcaaactcctgacctcaggtgatccacccaccttggcctcccaaagtgctgggattacaggcgtgagccact gcacccagccccttgtgtgtatttctgacagacaacccaagaaattacaggctgaaaccagaaaagaacacattaaagcaccaagag aaagttggagtgggttgaagggaaacagatttttaaagttaaggctctgtgaaatgggtagaattaactacaggttaaaaataaaatgtta actaaaggttgcctctgagtaacaggattatgggtgattttaattgtcttctttgtgtatgttcaacagtgactataatatgtattacttttggaat aaaggaaaacctgaaaggtgtgttgttttataagggcccttaggttgccaaaattagagtcattgaaatctaaagctgataaaaactttagt gcaaagattgtgacatgggagactacacataccagatccataatgtacatgaggacagtaggccgaggggccctgcacattgaaagc ccacatgggagaagcccttgggaaggggagtggaaggatgaggcaaggggccggggggatgcagaggctggcaggcagtcatt tctcagcttcagccattcccgccatgggggaatgtggacagagaagccaaacaaatctcctaaacagtaaatgtcagtcttctgtgtcag atatttaagaaaactaacagaggtcagagaagacacacctacagcaagtagactgtccctgtgctgcctttttgcaacccctgctttggc agtgctcaagcccacctcctgctctgtgcagacatctcttctttgctcttactagaccaaggtgaaagaaaactctcaccttctcccatctg gccccacagcatctggaacacactgatcctcataatccttgttcttgagaaatattaatgacttaatctcccaagcttgctccctctcctgtg caggccatctcagtatgttttgcagacaagacccagagaagtccagactggacttgttgcagactgcaaaactgccattggaaggcctc cgtcccagtccttctacagagtagccagtgggattcccagccatgggcgaggtgacagcagaggaggtggagaagttcctggactc gaatattggctttgccaaacagtactacaacctccactaccgggccaagctcatctccgacctccttggggccaaggaggctgccgtg gacttcagcaactaccactccccgagcagcatggaggagagcgaaatcatctttgatctcctgcgggactttcaggagaatttacagac agagaaatgcatcttcaatgtcatgaagaagctgtgcttcctcctgcaggcagaccgcatgagcctgttcatgtaccggacccgcaatg gcatcgcagagctggccaccaggcttttcaatgtccacaaggatgctgtcctcgaggactgcctggtgatgcccgaccaagagatcgt cttccctttggacatgggcatcgtgggccatgtcgcacactctaagaagattgctaacgtccccaacacagaggaggtactctcttcccc atgagagagagggcatggggcattattacatggagttctggggtacaggtggggtgaggggcattggccaccaagacaaggctggt gacaacatggtgcttcttctttatttatttacatatttactcataattctttattttgaaaaaaaaattcaaacttaacagaaaagtttgaaataata cagagaactgtatgtctttcatccaaatttatcattcattaacatcttgccacatttgctttctctctccattttctctgacccatttgggagaaag ttgcagacatcatgccctttgtctgtaagtactggcgatgctcccatcattgtactgtggataggtaagaaatgtccctgttctggaaatac acagtacttccacagtacttctacacagtacttccacagtacctccacagtacctccacagtacttccacagtacttccacacagtacctc cacaatacctccacagtacttccagaacaaggacatttcttacctatccacagtacaatgacggaatcatagaaatgtaatgtggatacg acaccattatctgctgggagtagtggcacatgcctgtagtcccatctactcggagggctgaggcaagagaatcacttgagcccaagag tttgagaccagcttgtgcaacatagggagactcctgtctctacaaaaaattttaaaattagctggatgtggtagtgcactgtgggggaaa gaaagagagatcagactgttactgtgtctatgtagaaagaaaaagacataagaaactccattttgttctgtactaagaaaaattattctgcc ttgagatgctattaatctgtaaccctagccccaaccctgtgcttgcagaaacatgtgctgtgttgactcaaggtttaatggatttagggctgt gcaggatgtgctttgttaaaaatgtgtttgcaggcagtatgcttggtaaaagtcatcgccattctccagtctcgagtacccagggacacaa tgcactgcggaaggccgcagggacctctgcccaagaaaccctgggtatggtccaaggtttctccccactgagacagcctgagatatg gcctcatgggaagggaaagacctgaccatcccccagcccaacacccgtaaagggtctatgctgaggaggattagtgaaagaggaa ggcctctttgcagttaagataagaggaaggcatctgtctcctgcttgtccctgggaatggaatgtctcggtgtaaaacccaatcatatgttctatttactgagataggagaaaactgccttatggctggaggtgggacatgctggtggcaatactgctctttactgcactgagatgcttgtg taaagtcaaacataaacctggcctccgtgcacatcaaggcacagcaccttttcttaaacttatgacacagagacctttgctcacatgttttc ccgctgaacctctccccaccattaccctatagtcctgccacatccccctcacctagatagtagagatagtgatcaataaatactgaggga actcagagaccagtgctggcacgggtcctccatatgctgagtgccagtcccctgggcccactgctctttctctagactttgtctctgtgtct tatttctttcctcagtctctcatcccacctgacgagaaatacccaggtatttctcatggaggggctggcccccttcagttcacacctgtggt cagcatcaatatggtgacttcccaggagtaagggaccaccaggctgcctaaggtggggtgaaccagcccaggtcagaaagggaac aggtcaaactctcatgctggttagtagtgggattgcacctgtgaatagacactgcactccagcctgggcaatatagcaagaaccgtaat gaagaggaaaaaatgaaaaaaggctgggtgtgatggctcacacctgtaatcccagcactttgggaggctgaggtgggaagaccactt gagcccagaaatttgagaccaggctgggcaacacagggaggtccctgtctttacaaaaaataaaaaaactcgctgggtgtggtggtg cacacctgtggtcctgtctactctggaggctgagatgggagcatcacttgagcccaggaggttgaggctgcagtgagccatgattgcg ccattgcactccagcttgagtgccagagtgtgaccctgtcaagaaaaggaaaggaaaggaaaagaaaggcaaggcaaggcaagga aaggaaagctaccatccaatcggctgggtgaaaaggaaaaaagaaaaaaattgctaaaaaataaagaaaaaaaagatgcaacacca tgatctaataaacagttcatattcagtttctgtcagttgtctcagacaccgatgtagcgctgtcttttgagtctccttgaatctgggactgttcc tcagtctgtctttgttttccacggtgctgacatttttgaagggtacaaatcaaatcttttgcagaaatgtcctttcatttgggcttgtctgatattt cttcataattagatgcaggtcacacatttccggcaggagcagcacagaggtgatgtgtttttctttgtgctttacatgcggaggcacaatg gcagtctgtcccattactggaagtgtaaacactgatcacttggttaaacacttggctctttcttccaggatccactgttccctgagattctcct ggtggttctgacaaagacttctttgatcaccttgggcaaatcactccccctctctggatgttggtttttaaaggcagtaatgcagttctccac atagccctctgcccaggccaactcctactcatccttcagaaaaagtcacccttcctcagggaggccctttctaatctccctgttatagccc cctccctaagccatttctcataataagccctggaggtcagggaccaggtctgccttgtccaccaggcttttactccaggacccagcaca gtgtctggcacaaagcatgtgcttggtaagcacttggtgtaaaagactcacaatcaagtaattatgcaataatttctttaacactgtctcacc cagaagaatgtttgtcccattacggcagggcccatgcgggtcttattcactcctacactggccaaaccaggagctgtgttccggcatttc caaaatccagcctttggacatggtgtgattttgtgctcattttacctctggagttacttcagattaagcatcttaaggttaaaaataggagag gaagggggcttcagagactgtcttatccaactgccttatttaatagacaaggaaactaaacctaggagagtgtgggcagaggcttgcca atgctccacatgcatgaatgcaagaattcaaacaggtaggccgggcgcagtggctcacacctgtaatcccagtgctttgggaggccg aggcaggcagatcacttgaggtcaggaatttgaaaccagcctggccaacatggtgaaaccccatctctactaaaaatccaaaaattag ctgggcctgggggtgcatgcctgtaatcccagctactcgggaggctgaggcaggagaatcatttgaacctgggaggtggaggttgca gtgagccgagaccgcaccattgcactccagactgggcaacaagagcaaaactgtctccaaaaaaaaaaggattcaaacaggtatcttt tggtcttgggctccttagtgagataagccagacttgtgcctctgtggaaagcatacccggtgggctcccacatgggcgcatggctcccg tcatctcctcttgggcatccaggggccgggcaatggttctccactcaggagactttccggaggactcatccaactggactggagtctatt ctcccttcaactcttctaactgtaattgtagccaacaaggattgcctcactcgtttcccctcataacgatctatgtcataactgtcattatctct gtttcacagatgatgaaacagaaacccagaaagttttggtaacttgctcaggattatgcagatagaaagtggcagatcctggctggttca agtacacaggtgtttataactaattgatcacagccagttatagatttctttgttccccctccactcctactgcttcactcgactagcctaaaaa aagaaaaaaggaaagaaaaaaaaaagtggcagactgggaatgatttgaactcaggcattctgaccttgagctcacaatcttttttttttttt ttaactagttggttcagccagggagtcgttacagagccttaattcctcttaattgctctgctattctgtactccctcctcctctataaaatgag ggcatcgcactgggtatttgagcaaggtcattggatcatgtagcagagagttaagatcttatgctttaaggtgagatctggatgtgaacca ggactctgtcactttcaaagaatgtggccttggcaaatcgtttagcttgactgggtttatttcctattttataaaatggggaaataagagctac cccacagcattgttgtagaaattaaatgaatgaattatctcatcagcagtgttcttggcacaccgtaatggctcctattggcagctcccgtt ccttattactattacgaccattttgctactgttattgttgtcgttgttaaggatctattctaagtgatggatgctgtaccaggtttgagtggaattt ggagaagaatccagtcttgcccttctggagctttcagtttagattctgtttgtgggaaaacagagaacataaatgctgagagcttctctata ccttggtgttaagggattgaggattcagggtgtttttctgagtctagaagatcctgacttaacagtttttggagcctctgtgctaagggacg gcctgttgctttctgttcatccgtcagagtttacccaggaaaccacacctcaccttccctggaaacttttggggcctgagtcagtgtccctg gccagtgacttgggcctgtgggtgaagtctgtctaggccagtcatccctgcgtacttggtatttgcacgtgtttacttcaggaaatcttact caattcatggccctgggcagaatgcaaccttgggccccagtggcagaatgcaaccttcgccaccaaacagtgactggggctgggtga aggagaaggaaggtggtgtagacaggggtctagattctttctggaccaccaaaaagtgatactctcaacctttagcaccctccaaaaac ctgagaaatggttgcaattgctgcattgttgaaatgcgaaatcacaaaggtagttcttttgggctgtaatgaacttccccaccccctccaaa ttcctacttatcttgggaattgccttgggaagatttgccacaggactgccagaaaaaatataagatgcccagtaaagtttgaattttagata aacaaagaatgatattttagttaaagaagatcctgtgcaatattttggacacagttacactaaaaaattatttatagtttatctgaaattcaaat ttcattgggaaggcccacttgccattcaatatttacttgtatatcattcctttgccctttctcctggcaggaaaacccatgaggtctgtctgtct gtccctgtcgatcttattcaccactggcccctggcacctagcccagtgtcttggacagagtgcatacttaccatagtttattgagtgagtga atgactaaatgtcagcccttcaccagcacttcctctggctaataaattccatttctgaaggctgtacctgagatgaccgtcatttgtagaccctctcacatttgttatctgcccctgctttccttcatttcccatgagacctgagacctagcaggctagaacaggaaaacaaagaaaaaaaaa accttcacagaggaccttattcaatactctgacttcagcaaaggggaaattgaggttcagagtgttttcaactactgacaatgtactgagt ggcccttgaagccactcataggacaagtttctccttatctgggtccccagcgtggtctgcttcacatgctttgagtcgtgggctcatgtttt gcatgacattccactataccatgagaacccactgagaacaacctcatcttacaggtctttgcatatacaatgcctggcatagtgagtgctc aatgaatatgaatgaatcaatctgtcaatcagtgaatcaatgaataagtcaatgaatgaataagtggagcaactgtgccacttaccagca agggagttgtctataaaatgggaaaggtggcagttctatttggcagggctgatgtgagagttaatgagataatatatgtcagttgtctggc tctgagcctagtcattattactcagtaaatagtagatctcatcatgagatgcaacagagcttggcactgtgaatgtttgcaaaggggaaat gaggaatctgggaatgagaagtagtgttgcattgcactttaaaatccaaactcagccaggtgcaggctgacacctgtaaccccagcact ttgggaggccgaggtgggaggatcacttgagcccaggtgttcaggactaccctgggcaacacagcaaaatcctagtgttacaaaaaa tacaaaaattagtcgggcatggtggtacgcccctgttgtttcagattctctggaggctgaggtgggaggatggcttgagctcaggaggt cgaggctgcagtgagccatgatggctacactgcactccagcctggatggcagagcaagactctgtctcaaaaataaataagtgaatac ataaaaatgaaatcaaaactctttcccatggcctccaaagccctgctggatctggcccctgcctgcctctttaacttccttgccctctgcttt ccctctccctctctatgctttagctacactgattttctttctgttcctcacgaggaccagcctcattcctacctgctggcctttgcatgctccgt gcttagaatgtgcttcatatacctggctcccttgatccttcagctcttagctcagcattacatcttcagagaaggctccctacccaccagcc tgccatccctttcatcactctctacagcaccatccagtccatttccttcaaagcagtttcctcaagttggaggaagaatgcctagttgtttact tgcttttttgtctgtctcgccgtttgaatgtaaggttcaccttatattacaacagtaatgttcattgctgtagtttcagggtctggaagagtgcct agcatatggaagacactctgataatgctaactgaatggatgaatcactttggtctttccctagctcagcctctgattctaggatttaaaataa atataagttggtgcaaaagtaatcgtggttcttgccattacttttactagtttgagctaaatggcttctaagtctccttctgcctttgataatctg agattctgtgacatggttgtttagaacactttatatgctgaatatatagggatgataatgatcatgtaaatgccagtcaattggcagttgaag tactcatgaggggagttccaggactgagtacccagtgggaatgagctttctggtggatttggtagcatctgccttggatgcaggtgagg cccaaatatttgcttttcctgaaagaagtgaacattgggaaactgaaatgacttcatgaaatgaatctgagcatactccacctagacacgt gctcaggaactcatcagtagcttagaaaatgacacagataccacttcttgactaatgcctttttagagattttgcttaatcctcaaatgttaat tcaagtacagcactacaagcttctcttaattcctttacccttcacctggttttaaacagcttcactgataataatgctgttgactatttatttaatc ccattccaactagaagaaatctatagatgcagagcattatgctctgaattccttccttccttccttccttccttccttccttccttccttcctttttt agagacagggtcttgctctgtcacccagggtaaagtgcaatggtacaatcatagctcactgcagcctccaactcctgggctcaagcaat cctcctgcctcagcttcctgagtagctggaactacaggcacatgtcaccacacctggctaattttttttgagttttagtagatacaagatatt gctatgttgcccaggctggtcttggactactgaacgcaagcaatactcccaccttggccttccaaagtgctgggattatagacgtaaacc accacgcctggcccagacttagttttagaaaagttctgtgaattggctctatgaagtacatttccactcagaacggcgtgagctgagcat gtttgggctggcccgcctcccaagcaatgctgacagtccaaaaacctcacagtgctaccaataaacagaaaggaggattcttgtaaatt taagagaacaattcaagctctgtcttttcatcctgaccgcatggtccaccacagcctataatggcttctctgggctgcattaggggaggttt atggaggtgtgtgctcctctatacactcagagcttcaagccaatcccttgtgacctacattactattattgatttttctttgagctggcctgaa ccttggtcacccatatcccccgtgcttaacataaggcctgacacatagtggggtttctactaagattcgtagaatggggatatctactgaa ctggttgtgtggtgctagaactggattgtttaaaaagctgaaaataacaaattcttttgttcaatgacctagacccaagtctaaccttatcca ggcttctctaagaggcagcagagctgaaggcaggctctggagttacacatacacacacacacacacactcacacacacagtctacac aatctacacacatgacattaaccttcttgagttacagatcccccttctgtaagatgggctactgatggtacttatctcatagcttattgcgaa gataaaagtaagataacgcaggtaaaatgccatgcacgctgcctgctagcaactgtcgttgtacttcctaaagtctttacccggcttcctc ttagctgtaccctgggtttttattctgggcttaattgtcatcacctcaagaaagccttctctaaccctccttctccaggcaaagtcagtccccc actcccatccctggaacccacactgctttgcacacctctcctccacagccctcagtggatttcatctcagactttcttcttctactggattttg gaagcttccatgacagatgtcatctctgatttccctgcccctgttgaagaggaatagatggaagcacctttattcccttgtcaacaggggc tttgggccgttccactgttcttgctcttcatttgcctgtccgtgcttatctctgccttcctttaggatgagcatttctgtgactttgtggacatcct cacagagtacaagaccaagaacatcttggcttcccccataatgaatgggaaggatgtggtggccataatcatggctgtgaataaagtg gatggatcccacttcaccaagagagatgaagaggtaagaatgctttagagactaaggatagccatgctagcacattctcccagcctatg aagtcccctgaactttgctgtttgacctggcctcagttttcccatctgtaagacagggaattcacctgatgttctctgagggaggcaggat aaaataatagttaagacattgggctttggagtcaggaatagagggataattgttctggctccgccaatagctgtttgccttttgcaagttatt taacttctttgaactttggttttcttatttgtaaaatgttgataatttttcagtatcaacctcatagggaacttattaataaatagtagctatttttcat gaatacactcccatcaacatcaacattctacattcggtgagttttatgctcatgcaacatcttatgagtctgcattctaggatccatctgccc aaattgtctctgatgaagctttgaaaatattcagatatttgctgatggtaaaagtagccagaggatggatttcttcttctcacctgcctcccct catgctgtttgatttttagattcttctcaagtacctcaattttgcaaatctaatcatgaaggtgtaccacctgagttacctgcacaactgtgaaa ctcgacgtggccaggtaaggggcttgggccaagggaatggacttgactcgtcctttccctgaagcattgatgtggtcaggactggcag gcagatgggaatgaaggtgcctaggaagccagtctctggccagcatcacgagtgttatgaggtagtggttgtctgtaaagctgtataccacagtggttatatgcacaagcttcaagacagatggctttgatctgaatccctgctcaaccactgcctagctctaagaccctaagtgagtca cttaatctctctatgcttcagtctccccatttgaaaaatagggatgattatggtgccaccctcaaaaggttgccatcaatagtaaacgagat aatgcatatagggcaatagtgctcaattgaagtttcttattatttttattaaaaacctatgagataaatgaaagggaaagagctggacccaa tgtaaagcagccagttacttcatctctctgagccccagtttgtttgtctacaaatggaagataattggactgggcatggtggctcacgcct gtagttccagcactttgggaggccaaggcaggaagattgcttgagcccagaagttcgagaccagcctgggaaacatagtaagacccc atctctatttttaaaataaaatttttaaaataaatggatgataatgggctgggtaaggtggctcatgcccataatcccagcactttgggaggc tgaggtaggaggattgcttgaggccagaagtggaaactcatctctacaaaaaatttaaagattatctgtgtgcagtggcgtccacctgta gtcccagctgctggggaggctgagatgggagaattgcctgagcccgggagttggagactacagtgagctatgatcgtgctactgtact ccagcctggatgacagagtaagactctgtccctaaaaaataaatacataaaaacaaacactttagcttttgcgcttctgccctggccctct gctggccactccgccggtgctgtgcctgggcgcctccgtgctctcggccaactgcctctgagagcgcccacttgagcgccctgggag ccggagggcggcggtcctcaccgggaccctcctgtgggcggagggggacaaaagtgtctctcaatccagcacatgcacattgaag caagttaaaggatttaatatgaagcacagaagccgacagtgccaaacagcaagcagtagttggtacacatttggtgagtggggcagc atttccttctcccactgctgctgagatggcagaaattagtcgaattcgtacgaaatggaatatactgaaggcattagtcagcgaatgagg gtcccagaaaagttaaaagtagcaccgccaaacgccaacctggaacagggattccaaaaaggagttccaaatgctagcgtgataata atgcaagttccagagaggattgttgtagcaggaaataatgaggatgtttcattttcaagaccagcagatcttgaccttattcagtcaactcc ctttaaactcctcgcactaaaaacagcacctcgtgtacttaggctaagtgaaagaccactagattttctggatttagaaagacctcctaca acccctcaaaatgaagaaatccgtgcagttggccgactaaaaagagagcggtctacgagtgaaactgctgttcgccaaaatggacag ctggtcagaaatgattctctttatggcatttcaaatacagatacaacaattgaaggaacatcagatgacctgactgttgtagatgcagcttc actaagacgacagataatcaaactaaatagacgtctacaacttccggacgaggagaacaaagaacgtgctaaaagagaaatggtcgt gtattcaattactgtagctttctggctgcttaatagctggctctggttttgccgctagaggtaacatcagctctcaaaaatattatctcaacag ctggaaatataaaagatttgcaaacttctttgtttctgtctcttcattgtacgccattttatagtccacaacctgaaaatatatttcttccagaaa ctctggaggtaacacctgcttaaaattctccttttgcatgttttgtaaataggctccagttttgttttttaaaaggaattcatcttttgcctcatcg gtccacacaactgattctgaatgggagagagtctgtagagaattgatttagaaaaatgtctgtgaaagaaaaacaattattttgtcgtgttt ctcaaacactgtaagcagttttgttaatagacatttttccatcaacacttcaacattaacactttcaaagtcatggtctgttgccagatttgaga aactcgaaccacctaatatttcataaccttcttcattaggtacttgtacagattaatttctaacatttcagcagtttaatatgtgtgcaataagtg cattctttcattttagttttgtacttggttttctataaagtacgtttttactcagttcatgtgtgaacaatttttaaaaactacagaataaggtacaa atgtagtgtatttaataaactgtcaaccaaagcaaaaaaaaaaattaaaaactttaaaaaaatctaaaagtaaaaataggggatagttaac ctgtctatttctctggattattgtgaagggtaaatgaaataaactccagacaactttacagaattgttgttattctccagctaagtgagctact actttgcacctcaaaacctatgaatatgtttttgtgttttttcagatactgctgtggtctgggagcaaagtctttgaagaacttacggacatcg aacgacagttccacaaagccctgtacacagtccgtgctttcctcaactgtgacagatactctgtgggtctcttagacatgaccaagcaga aggtgagaaaacttcagcaacagacatttccaagaaaagattaaacagacactgactgacggctagagtcttggcacacattcaattgc acgggggaagaggctgggtgaattgggatggttgtagaaagtcttacataggtctctactgaggagtatcagcctttaaaggggaaaa gtcaatctagggcacaggtgagtgaaaacaatctgtactttccttagtcatttggaaagaaaaaaagtgtacatgaaaagaaactgatga ggtgatgaaggccagtgataaagaaatttctatgtgggttgagagtccaggagaaagttgccaacttcagggattctcaaaaaatgtact gaaagttgtgtgtaaagctttgctgcaatttatctattccagagctcacccaggcaattaagtggaggcagatgccgacatcgggtccct cacatgttcaggggccagatttgtggcagatacagcagctaaggaggcagagttcgatgtcgctcccagcgatgttttttagatatgtttt cccaaacaacttcctcatttctctttgtttatttgcttctgccttatgaggctaatcattttctgccttgaaacagagtttcagagttatctaagttt aatatctcctatcctgaaaacccaaggattccagccagtttcattttgtacgccttgtcatgctgagggtggtgcccagaccaatgcagac tctccaaatatattggattgagtgaatacatggatcacagaatctggggttggaaaagcccttggatatagtccagaggctcccaaactt ggcagttcatcagaatattctgaagagctgattaaaaaattactggactgcattttctgatagtctgccattttttacttaatgttttacaaacat ctttccatgtcagcacataggaattctttttaatgagcctttatattgaaacataggcctggcacagtggctcacacctgtaatcccaggact ttaggaggttgaggtgggaggatggcctgtggccaggagttcgagtccagcttgggcagcatggtgagacctgtctctacaaaaaata aaataaaaataaaaaaagaaggacaacgtagacacagaaaagggacattaattatagctgctgatataacagttcaatgagtgaacatt cccatgaaaccaccaccaaaattaaggaacagaacatgacaaacagcacctcagcagcctccttcatacctttttcagtacctcccaca gcctcctcctgaaaggttaccaccatctgacgtttaacaccatattagttttacctttttgaggttcatatatatgaaatcctacaatacacatg cttttgtgtttggcttcttttatttatttatttatttatttatttatttattttttgagacggagtctcgctctgtcacccaggctggagtgcagtggcg caatctcggctcactgcaagttccacctcccgggttcacgccattctcctgcctcagtctcccgagtagctgggactacaggcgcccac caccatgcctggctaatttttttgtatttttagtagagacggggtttcactgttttagccaggatggtctcaatctcccgacctcgtgatccgc ctgcctcggcctcccaaagtgctgggattacaggcgtggtgtttgacttcttttgattcattcttattacttcatttagcatgagttcattcatttt tattcctgtggaattttccatcatatgtacaatctacagtgtatttattcatttccactgttggtggacatttgagttttcagatctgatctattattaatactgttattatgaaacttctagtatattactagtacacagttgtgccctttctgttaggtatttacctagaagtagaatttcagagtattagctt tagtagatattgcccatctgattttcaaaatgattgtaccattttataccctcaccagcaatgtatcaagggtttcttcatcaatacttggtatta ttattcttagtaactttagccattctggtgagcatgtggctttatctcttttgttatcaacataaattcccctaatgactgatgaggttgaatatttt aaaatacgtttattggccatttggatgtattattttttgaagtgactgttcaagtttctcctccatttaaaaaaatcacattgtcttttttatctcact gatttgtaggagttctttgcaaattctgggcacaagtccactttcaaatacatgcattgcaaatacagtcacacaccatataacaacattttg atcaatgatggcccgcctataccatggtcccatagaattataatggagctgaaaaattctatcgtctagtgatgtcatagtgcaacgcattc ttcacatttgtggtgatgctggtataagaaaaacctaccgcactgccagttgcataaaagtatagcacatacaggccgggcgcggtggc tcacgcctgtaatcccaacactttgggaggccgaggcaggcggatcacgaggtcaggagttcgagaccagcctgaccaacatggtg aaaccccgtctctactaaaaataaaaaaattagctgggcatggtggtgggtgcctgtaatcccagctactcaggaggctgaggcagga gaattgcctgaacctgggaggtggaggttgcagtgagccgagattgtgccactcactgcactccagtttgggtgacagagtgagactc cgtctcaaaaaaaaaaaaaaaaaaaatatatatatatatatatatatagcacatacaattatgagctgtagtacataatgcttgataatagaa aactagaaaactatgttactggtttatgtatttactatactacaacttttttccattattttagagttgactccttctacttattaaaaaagaaaaaa gttaactgtaaaacagcctcaggcaggtccttcaggagacattctggaagaaagcattgttatcataggagatgggagctccatgcgtg tttttgcccctgaagacgttccagggggacaaagtgtggaggtggaagatggggatattgatgatcctgaccctgtgtaggtctaggct aatgtgtgtgtttgtgtcttagtttttaacaaaaaagtttagaaagtaaaagaaaaaaaaatttaagagaaaaaaagcttgtaggataaaga tataaagcaaatatttttgttctgattcaagcaccaaagacataaaaaaagaaaaaagaataacagaatttgagatttttttaaaaagaaaat atttttgtacagctgtataatgtgtttgtgtattaagctaagtgttattacaagagagtcaaagtttttaaaaataaaaaattttataaagaaaaa tttaaaaatttatagagtaaaaaagttacaagccaggcttggtggctcatacctgcaatcccagctactcaagaggctgcggtagaagaa tcgcttgagaccaggagtccgaatctgcagtgagctatgatgagcaagatctcatctctaacataattaattaattaattaaaaattacagt aagctaaggttaatttactattgaagaaagaaattttaaaaatacatttggtgtagcctaagtgtccagtgtttataaagtctatagtagcgta cagtaatgtcctaggccttcacattcactcaccactcactcactgactcacgcagagcaactctcagtcccgcaggctccattcatgcta agtgccctttataggtgtatgattcttaaacttttataccttatttttactgtgttttttatgtttagctatgtttaggtgtgtagtaggctatacaatc tggagtaatgcaagtgcactctgcaatgttcacacaatgacaaaattgcccaaagaaacacacatttcctggagcatatccccgccatta agtgacgcatgactgtattctcccacttggtgttttcattctctctctctctctttttttttttttttttttggagacggagtttcgctcttgttgccca ggctggagtgcaatggcacgatctcagctcattgcaacctccacctcccgggttcaaacaattctccagcctcagcctcccaagtagct gggattacaggcccccaccaccacacccagctaatttttgtgtttttagtagagacagggttttactatgttggccaggctggccttgaact cctgacctcgtgatccacccacctcagtctcccaaagtgctgggattataggcgtgagccactgcacctggcccatgctttcattctctta atggtgtgtttttatgaatataagttctgaatgataatgtagttcaatttatcaatcttttcctttatagtcattgattcttgtgctgtaatgaggaa atctttcctggatctaagattgtagagatatttacatatgtcaaattctaaaagctttattgttttaactttcacatatagctctacaatctacctg gaacttttttatgtttagtgtgaagtaatgtgagtattcatttttctgtacaagcatccaattggcccagcactgtttattgaaaagaccattcttt ccctactcttctagtgaaaagatcatccttccctatcatacctttattatcaaccaagtgtccatatatgtatgggtctgtttctggattctctatt ttgttccattggtatatatatatgttttttttaattaatttttttttcaagacagatatctcactcttttgcccagcgtggagtgcagtggtgcgatttt ggctcactgcaacctccaactcccaggttcaagcaattctcctgccttggtctcccgagtagatgagattacagacacgtgccaccatg cccagctaatttgtgtatttttagtacagatggggtttcaccatattggccaggttggcctcgaactcctgatctcaagtaatctgcccacct tggcctctcaaagtgctaggattacaggtgtgagccaccatgcctagcctccattggtatatttacctattcttctgcaatactatcctgtctt aatttcagtatgtttataaaatatcatgatatctggtaatgaaaagacctctaagtttattattattatatgttagactttttcaccatggtttgtgtc tcttatacttctttgtgtgtttttcatcatttcgttttttgtgctttagtttgaatatttcttcgatccatcttctattaattactaactctttttttttttttttttt gagacagagtctcattctgtcacccatgctggattgcagtggcatgatcttggctcactgcaacctccgcctcctgggttcaagagattct catgcctcagcctcctgagaagctgggactatgagtgcaggccaccatgctcagctaatttttgtgtttttagcacagatggggtttcgtc atattggccaggctggtctcgaactcctgacctcaagtggtctgcctgtcttggcctcccaaagtgctggtattacaggcgtgagccact gcctggctaactctctcttcaacagtgtctaattgactgttaaactattgagtttttaactttagatatttttcaattttagaatttccaactcattct gtttttgtagtttccagttctctgctaaagttctctgttttgtacttgattcttggacataatcacagttatttttgagtctctgtctgaaagctataa tatttataggcatcttctatggacctatttctatgatctgtttcccctctagatttttccgtgaattgtctgttacatgctttattatttttgtttgaaga ctagagacatgtatggaaatatgttatcttcctccaaaggggatggttttacttctggcaagcagttggggtaaaggcagagaacctcaat ccatttaggaattgaactgatgtgaagctggtcttcagtcttcgtgaggcctggtctgcttctggttcacttttacttcttggggcgtgaggat cccagcctagggtgttttcaagggtctctcctccttggcaggctctgaattccaattttgtcctccttgcccagtgagtcttccgtaagcttg gctcagcttcttagctgctcacagcatctttggtatcaacaagtgggtcaaaaggaaaggtcgaatcaagggtcaggtttacatctctgg gtttcctttctctctgggattttggcccactaactccctgctgccttgccggctatataagtccttcacataaacaaatgttcatacacataca cacagtccagatggtgtaatttttccagttggaggattggtctgaaacaaacgagtcatcatgaacagaagtaggaatcctctctctttgtt ttgtagtccatatggatttatctcattcatttgagtagtataaatataccgtaattaatttatcctttcccctattgctagacattaaaaattatttctaaatattcttataactaaatatatatatatttctatttctaaatattcttataactaaatatatatatatctatttctaaatattcttataactaaagaca atgctggccatacaaccccttgtttgacaacttctcttctccttgggacaatctttggagccactcagtgtttggcccacatgaggtgtcag gccaatatatttgttcttcttctgggctcctctagcatttatcctctccttccagcactgttttggttcttgcctcatttacctcactattagttatctt cttcatgagacaataagttcttccaggatacaaactgtgccttattcgcctgtgtataactagcagtcagcaggttgtggggctcaggtag ggcctggttatatttttgcctagaattaaaattgttttctaaatgttcacctagaattaaatttcttcctgttgttggtagtctagaaaaagagctc aactggggagaagggcaaaaacatggactctgcatttcatgtggcttcccttctgcttctctggatgatattagaataagcagaccatga caaacatgttcaaatgagaggaatgaggagggcttggcatcctacccacagtaacccagtgctaatagatcacggtttgtccctgtata gtaacagatgaatgaacagaaagggcactggaggatggaagcccctctctcaggcccagacttgaccccacacaagctttgctgctc tatgggcaaaggaaaccatgaagaagtagtggggaagaacctgtgtggttctcagttggcctctcctgaggctgcatgactgtgcagc tcctttgctggctctttttcctcttccctctgcctgtgtcttggccacttttatttagtgttgttgtttgttatgttatgttatgttatgttatgttatgtta tgttatgttatgttatgttatgttatgttatgttatgttatttatttgagacagagtctcactctgtcacccaggctggagggcagtggcacaatc tcagctcactgcaacctccccctcccgggttcaagtgattctcctgcctcagcctcctgagaagctgggattacaggcacataccacca cgcccggctaatttttgtatttttggtagagacggggttttgccatgttgctcaggctggtgtcaaactcctgggctcaagtgatccatccg cctcagcctcccaaatggctgggattataggtgtgagccaccgtgcccgaccttggccacttttagacaaaacaaattcaacagagttta tatgagcaacgaacaactcatgaattgggcaaccttcaaaaccagaagaggttcaaagagctctgctgcacagcaaggccagtgagc tttcatagctgaacatggaagtaaagtatagaaataacatgattagttacaatagggcatttgtctgatttggacatgatctgatcagttggg ctgcctgtgattggttgaaactcgaccatttgttacagtaatatactcctaaattgggttttggtttgtttgcctactgaggttgcagtttcttttg taggagctcaaattattacatggaaactggaagaacagagatagcctcaggctaaggcacctgtgtattttgctttagcagccacttatct tacaggtgtcagcctttcttattggatcccagatataggtctattgtccatatttgatatcatttttttctttcttatttgtgaaaaatattttttcctg attacaaaagcaataaagtttgttatgaaaaatttctaaataaattgcagtctgattccaatcactcagagaaaatcacttgttaatactttgtt gtgggccaggcatggtggctgatgcctgtaatcccagcactttgggaggctgaggcagatggatcacctgaggtctggagttcgaga ctagcctggccaatatggtgaaaccccatctctactaaaaatataaaaattagccggatgtggtggcacacacctgtaatcccagctact tgggaggctgagccaggagaattgctagaacccgggaggtggaggttgcagtgagctgagatggagccactgcactccagcctgg gcaacagagcgagaccctgtctccaaaataataataataataataataataataatttgttgtaaatgtattcatgacagtttctatatataatt gaacattagggtcacatgatatattctttttggcctctcacatttgtcacttatcagtgtatcatctttccaatcagcacacacacgtctgtaac atcattgttaatagatgtaccataattgatttaataagtcctttcttcttgggcacttatattgtctctaatttttcactactataaatgatactgctt gggacattgttttagctaaatctttgagcacatattagcatggtcttaaggtaaatttcttagccgtaaagaccctgctttccctctcatgtcc aataacactgagggtgccccagttcagctcccatgtttttggctaatgactagacaggcccagttcatcaatattttgaagggcaaatgtg actattttctaactgtagtgagtcttactgcctttccaagtttaggtttcactgagttcttctcactttctaaatttcagtgttagctgactcatgga ggtgggacatcttaatctcctttccttgatctcttttcaaaacatgtaattattgtttttctcattcctaggaattttttgatgtgtggccggttctg atgggtgaagttccaccttactctggtcccaggactccggatggaagagtacgttctctctgaccttttcagtatgatttaatatattcaaat gcaaggcacagtaaaccctgttaaaggattttttattagcatataggtatttgtctttgcgttgggttgtctcctttgaagttagaataatctctc atattcaatagcagacaggtgctaaaagtaaaataaaacaacagagagaaaaaaagtaaaaaaaaaacagcaacaataattcctcata attgtcatgccctaacatccctaacatttgcaaagtggtcttccttccttccttccttcctgccttcctgccttcctgctttccttccttccttcctt tttctttctttctttctaagcactatactaacaagaagctaagtgtttaatatcttattggggtttcacagtaaccttgcggggtagaccaaaca gggaccaacatactcatttcacagctgtggaaagagacccagaaaggtcaagcaacctactcaaggcttcaccttgcaggaatttaga gttatagacaatatgagcttagaaaaccttcatatctagtctataatgtacaaattggtagccagaaagccaattccagctgacagataca cttgcttggctacgtaggttttctattttctttctttttctttctttctttctttctttctttctttctttctttctttctttctttctttttctctctttctctctttctt ctttctttcttttcttctctctctctctctctctgtctctctctttctctctcagatagggtctttgtatgttgcctaggctggtcttgacttcctagtgc tcaagtgatcctcctaccttggcctcccaaagtgctgggaaattggcggctaacccttaaaaatcaaagtctttccataaatacctgaattc tcagtttcttttgaaaaggtggatgattaatccatactggacccacaatccagcagggcaaatttgattggagctgagtagcagctacctc cttgggatgtggcctgagttctctagtttgccacagtccccgccactttctctttttgagatggagttttgctcttgtcacccaggctggagt gcaatggcgtgatcttggctcactgcaacctctgcctcccgattcaagtgattctcctgcctcagcctcccgagtagctaggattacagg cacctgccaccatacctggctaattttcgtatttttagtagagacagggttttgccatgttggccaggctggtctcgaactcctgacctcag gtgatccgcccacctcggcctcccacagtgctgagattacaggtgtgagccactgcgcccggcccccaccactctcttttatgtacatct gtgccctgctttcatttttgtttttatcaacctctctaggcatttaagtttctgatctctgatctcatccaattgcagcattttattttatttgtttttgtt ttttatatagagatgaggtcttgctatattgcccaggctggtcttgaactcctggcctcaagtgatcctcctgccttgagctcccaaagtgct gagattacaagcatgagccaccgcacctggcctaaattaggccctttaacattccctctgccttctaacatccctctcacgctttatctgcc acattcacttggaaacacagcttttgtctgaattccaaccaattaatgttcccgaatcttaggtcgaaagcctcacagtgtcagctacatctt gcttcatctacttttaaaaataaaatagtgtgtgtatgttttaaatatttaaaagttatacaggctaaaaagccttcatggaaaaaaaaatagcagtcttttaaaaagtggtgtatttccatgggtatctgtggctctttcaaagccccttgtgtcagctgctcaggtagaaggtaacaaaggtact cttgatgactcagttttgactttccagaggctggtgaatttgtacatgaatatggctttaaagactttgactgagatacagtgagtggaaaaa gcctagttacccaaagcacactgatctttttgttaattctgggaaagaaaaactttgcagtttttctcctgtgggattccaaaaaaaatgtcc atttctttttttgagatggactgtcactctgttgcccaggctggagtgcagtggcatgatctcagctcactgcaacctccgcctcctgggct caagcaattctcctgcctcagattcccgagtagctgggattacaggtgccccaccattatgtctggctaatttttatatttttagtagagatg aggtttcactatgttggccaggctggtctcagactcctgacctcaagtgctctgcccacctccgcctcccaaagtgctaggattacaggc gcgagccaccgcacccggcctagaaatgtccatttctaacctttctaaagactctcgtgggaatccctgggtaggacagcagccaggc ccaggaagccctgagcctgccaggagtacagtgagcagcacaggcccaaggacaggtggccctaaggcaagacaaagatgtcaa gaaggtcctggaactctgcctcctccccctcgcaggcagaaacttatctcctaggaaataagtgttctagaaagcagccacctttgctttc tagaactgcttatagggtacaacttgactatttaaaaatatttttaagaaagtttgttgtgccaggtgcagtggttcatgcctataatccaaga actttgggaggctgaggcagaaggatcacttaaggccaggagttcaagacctgcctaagcaa...

Claims

CLAIMSWhat is claimed is:

1. An expression construct comprising a rod-specific promoter, wherein the rod-specific promoter consists of a sequence comprising:a) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 18 and (ii) a sequence that is at least 80% identical to SEQ ID NO: 17; or b) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 19 and (ii) a sequence that is at least 80% identical to SEQ ID NO: 17.

2. The expression construct of claim 1, wherein the rod-specific promoter comprises: a) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 18 and (ii) a sequence that is at least 90% identical to SEQ ID NO: 17; or b) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 19 and (ii) a sequence that is at least 90% identical to SEQ ID NO: 17.

3. The expression construct of claim 2, wherein the rod-specific promoter comprises: a) SEQ ID NO: 18 and SEQ ID NO: 17; orb) SEQ ID NO: 19 and SEQ ID NO: 17.

4. The expression construct of claim 1, wherein the rod-specific promoter comprises: a) a nucleic acid sequence that is at least 80% identical to SEQ ID NO:8; or b) a nucleic acid sequence that is at least 80% identical to SEQ ID NO:9.

5. The expression construct of claim 4, wherein the rod-specific promoter comprises: a) a nucleic acid sequence that is at least 90% identical to SEQ ID NO:8; or b) a nucleic acid sequence that is at least 90% identical to SEQ ID NO:9.

6. The expression construct of claim 5, wherein the rod-specific promoter comprises SEQ IDN0:8 or SEQ IDN0:9.

7. The expression construct of claim 6, wherein the rod-specific promoter consists of SEQ IDN0:8 or SEQ IDN0:9.

8. An expression construct comprising a rod-specific promoter, wherein the rod-specific promoter comprises a nucleic acid sequence that is at least 80% identical to any one of SEQ ID NO: 10, SEQ ID NO: 4, or SEQ ID NO:3.

9. The expression construct of claim 8, wherein the rod-specific promoter comprises a nucleic acid sequence that is at least 90% identical to any one of SEQ ID NO: 10, SEQ ID NON, or SEQ ID NO:3.

10. The expression construct of claim 9, wherein the rod-specific promoter comprises any one of SEQ ID NOTO, SEQ ID NO: 4 or SEQ ID NOT.

11. The expression construct of claim 10, wherein the rod-specific promoter consists of any one of SEQ ID NOTO, SEQ ID NON, or SEQ ID NON.

12. An expression construct comprising a rod-specific promoter, wherein the rod-specific promoter:a) comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 1, wherein the rod-specific promoter comprises no more than about 800 contiguous nucleotides from SEQ ID NO: 13;b) comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO:5, wherein the rod-specific promoter comprises no more than about 400 contiguous nucleotides from SEQ ID NO: 15;c) comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO:6, wherein the rod-specific promoter comprises no more than about 800 contiguous nucleotides from SEQ ID NO: 16; ord) comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO:2, wherein the rod-specific promoter comprises no more than about 400 contiguous nucleotides from SEQ ID NO: 14.

13. The expression construct of claim 12, wherein the rod-specific promoter:a) comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NOT; b) comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO:5;c) comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO:6; ord) comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO:2.

14. The expression construct of claim 12, wherein the rod-specific promoter comprises any one of SEQ ID NO: 1, SEQ IDN0:5, SEQ ID NO:6, or SEQ ID NO:2.

15. The expression construct of claim 12, wherein the rod-specific promoter consists of a nucleic acid sequence that is at least 80% identical to any one of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:2.

16. The expression construct of claim 12, wherein the rod-specific promoter consists of a nucleic acid sequence that is at least 90% identical to any one of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:2.

17. The expression construct of claim 12, wherein the rod-specific promoter consists of any one of SEQ ID NO: 1, SEQ IDN0:5, SEQ ID NO:6, or SEQ ID NO:2.

18. An expression construct comprising a rod-specific promoter, wherein the rod-specific promoter comprises:a) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 18, (ii) a sequence that is at least 80% identical to SEQ ID NO: 27, (iii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 28, and (iv) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 29;b) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 19, (ii) optionally SEQ ID NO: 27 (iii) a sequence that is at least 80% identical to SEQ ID NO: 28, and (iv) a sequence that is at least 80% identical to SEQ ID NO: 29; c) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 19, (ii) a sequence that is at least 80% identical to SEQ ID NO: 28, (iii) a sequence that is at least 80% identical to SEQ ID NO: 27, and (iv) a sequence that is at least 80% identical to SEQ ID NO: 30; ord) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 31, (ii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 28, and (iii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 32; ore) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 36, (ii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 21, and (iii) optionally the nucleic acid sequence CGCGGCCGC.

19. The expression construct of claim 18, wherein the rod-specific promoter comprises:a) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 18, (ii) a sequence that is at least 90% identical to SEQ ID NO: 27, (iii) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 28, and (iv) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 29;b) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 19, (ii) optionally, a sequence that is at least 90% identical to SEQ ID NO: 27, (iii) a sequence that is at least 90% identical to SEQ ID NO: 28, and (iv) a sequence that is at least 90% identical to SEQ ID NO: 29;c) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 19, (ii) a sequence that is at least 90% identical to SEQ ID NO: 28, (iii) a sequence that is at least 90% identical to SEQ ID NO: 27, and (iv) a sequence that is at least 90% identical to SEQ ID NO: 30; ord) (i) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 31, (ii) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 28, and (iii) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 32; or e) (i) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 36, (ii) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 21, and (iii) optionally the nucleic acid sequence CGCGGCCGC.

20. The expression construct of claim 18, wherein the rod-specific promoter comprises:a) (i) SEQ ID NO: 18, (ii) SEQ ID NO: 27, (iii) SEQ ID NO: 28, and (iv) SEQ ID NO: 29;b) (i) SEQ ID NO: 19, (ii) SEQ ID NO: 27, (iii) SEQ ID NO: 28, and (iv) SEQ ID NO: 29;c) (i) SEQ ID NO: 19, (ii) SEQ ID NO: 28, and (iii) SEQ ID NO: 29;d) (i) SEQ ID NO: 19, (ii) SEQ ID NO: 28, (iii) SEQ ID NO: 27, and (iv) SEQ ID NO: 30; ore) (i) SEQ ID NO: 31, (ii) SEQ ID NO: 28, and (iii) SEQ ID NO: 32; orf) (i) SEQ ID NO: 36, (ii) SEQ ID NO: 21, and (iii) CGCGGCCGC.

21. The expression construct of claim 18, wherein the rod-specific promoter comprises:a) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 22; b) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 23; c) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 24; d) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 25; or e) a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 26.

22. The expression construct of claim 18, wherein the rod-specific promoter comprises:a) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 22; b) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 23; c) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 24; d) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 25; or e) a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 26.

23. The expression construct of claim 18, wherein the rod-specific promoter comprises any one of SEQ ID NOs: 22-26.

24. The expression construct of any one of claims 1-23, wherein the expression construct comprises a transgene operatively linked to the rod-specific promoter.

25. The expression construct of claim 24, wherein the transgene encodes a therapeutic protein.

26. The expression construct of claim 24 or 25, wherein the transgene encodes ArchT, R9AP (Rgs9-anchor protein or RGS9BP), Jaws (cruxhalorhodopsin), iClC2, RHO (Rhodopsin), GNAT1(G Protein Subunit Alpha Transducin 1), CNGB1 (Cyclic Nucleotide Gated Channel Subunit Beta 1), APOE (apolipoprotein E), HTRA1 (HtrA serine peptidase 1), or C3 (complement 3).

27. The expression construct of any one of claims 1-26, wherein the expression construct further comprises a post-transcriptional regulatory element.

28. The expression construct of claim 27, wherein the post-transcriptional regulatory element is a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE).

29. The expression construct of claim 27, wherein the post-transcriptional regulatory element comprises a nucleic acid sequence that is at least 80% identical to SEQ IID NO:11, 38, or 39.

30. The expression construct of claim 29, wherein the post-transcriptional regulatory element comprises a nucleic acid sequence that is at least 90% identical to SEQ IID NO:11, 38, or 39.

31. The expression construct of claim 30, wherein the post-transcriptional regulatory element comprises SEQ ID NO: 11, 38, or 39.

32. The expression construct of any one of claims 1-31, wherein the expression construct further comprises a polyadenylation signal (pA).

33. The expression construct of claim 32, wherein the pA comprises a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 12.

34. The expression construct of claim 33, wherein the pA comprises a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 12.

35. The expression construct of claim 34, wherein the pA comprises SEQ ID NO: 12.

36. A vector comprising the expression construct of any one of claims 1-35.

37. The vector of claim 36, wherein the vector is a non-viral vector.

38. The vector of claim 36, wherein the vector is a viral vector.

39. The vector of claim 38, wherein the viral vector is selected from the group consisting of adenoviral vector, adeno-associated virus (AAV) vector, and lentiviral vector.

40. The vector of claim 39, wherein the viral vector is an AVV vector.

41. The vector of claim 40, wherein the AVV vector comprises inverted terminal repeats (ITRs) derived from AAV2, AAV5, AAV8 or AAV9.

42. The vector of claim 41, wherein the AVV vector comprises ITRs derived from AAV2.

43. The vector of any one of claims 40-42, wherein the AVV vector comprises capsid proteins derived from AAV5 or AAV7m8.

44. A cell comprising the expression construct of any one of claims 1-35 or the vector of any one of claims 36-43.

45. A pharmaceutical composition comprising (i) the expression construct of claims 1-35 or the vector of any one of claims 36-43 and (ii) a pharmaceutically acceptable carrier.

46. A method of expressing a transgene in a rod photoreceptor in a subject in need thereof, the method comprising administering to the subject the expression construct of claims 1-35, the vector of any one of claims 36-43, or the pharmaceutical composition of claim 45.

47. A method of prolonging the survival of cones that otherwise degenerate due to a bystander effect in a subject in need thereof, wherein the method comprises administering to the subject the expression construct of any one of claims 1-35, the vector of any one of claims 36-43, or the pharmaceutical composition of claim 45.

48. A method of treating an ocular disorder in a subject in need thereof, the method comprising administering to the subject the expression construct of claims 1-35, the vector of any one of claims 36-43, or the pharmaceutical composition of claim 45.

49. The method of claim 46 or 47, wherein the subject has macular degeneration.

50. The method of claim 48, wherein the ocular disorder is macular degeneration.

51. The method of claim 49 or 50, wherein the macular degeneration is dry age-related macular degeneration (AMD), wet AMD, or geographic atrophy (GA).

52. The method of claim 46 or 47, wherein the subject has a rod dystrophy.

53. The method of claim 48, wherein the ocular disorder is rod dystrophy.

54. The method of any one of claims 46-53, wherein the subject is a human.

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