PORE monomers and their uses

Abstract

The invention relates to novel pore monomers which have been elongated to increase the distance between the upper surface of the pore monomer and a constriction in the pore monomer. The invention also relates to pores formed from the pore monomers and their uses in analyte detection and characterisation.

Description

[0001] PORE MONOMERS AND THEIR USES

[0002] TECHNICAL FIELD

[0003] The invention relates to novel pore monomers which have been elongated to increase the distance between the upper surface of the pore monomer and a constriction in the pore monomer. The invention also relates to pores formed from the pore monomers and their uses in analyte detection and characterisation.

[0004] BACKGROUND

[0005] Nanopore sensing is an approach to analyte detection and characterisation that relies on the observation of individual binding or interaction events between the analyte molecules and an ion conducting channel. Two of the essential components of analyte characterisation using nanopore sensing are (1) the control of analyte movement through the pore and (2) the discrimination of the composing building blocks as the analyte is moved through the pore. During nanopore sensing, the narrowest part of the pore (also known as a constriction) forms the most discriminating part of the nanopore with respect to the current signatures as a function of the passing analyte.

[0006] The movement of polynucleotide analytes through the nanopore is typically controlled using an enzyme, such as a helicase. Suitable enzymes are disclosed in WO 2013 / 057495, WO 2013 / 098562, WO 2013 / 098561, WO 2014 / 013260, WO 2014 / 013259, WO 2014 / 013262, and WO 2015 / 055981 (all of which are incorporated by reference in their entirety). In a similar manner, the movement of polypeptide analytes may be controlled using a polypeptide handling enzyme. The enzyme controlling the movement of the analyte through the nanopore interacts with the pore. The enzyme is typically too large to enter and pass through the channel of the nanopore. The enzyme typically physically or sterically interacts with the surface of the nanopore and is thereby prevented from entering the channel. This is advantageous because it prevents the enzyme from blocking the nanopore. There is a need for new combinations of nanopores and enzymes for use in analyte characterisation. SUMMARY OF THE INVENTION

[0007] The inventors have surprisingly shown that it is possible to modify the constituent pore monomers of an oligomeric pore to modulate the distance, such as increase or decrease the distance, between the surface of the pore which interacts with an analyte binding protein and the constriction in the pore. Pore monomers modified in this way and pores formed from them have numerous advantages. In particular, pores formed from the modifired pore monomers of the invention are advantageous for analyte characterisation because they demonstrate one or more of an increased peptide or polypeptide read length, an increased read length for any analyte conjugate where the peptide or polypeptide is swapped for something else (e.g., sugar), one or more improved signal metrics, such as an increasedsignal-to-noise ratio (SNR), an increased range, an increased signal steppiness and a reduced noise, an improved pore insertion into membranes, an improved electroosmotic flow due to more surface area to add charge density, an increased capture of analytes, a lengthened readerhead (if the analyte binding protein is considered as a readerhead), and an increased pore stability.

[0008] The invention provides a modified actinobacterial pore monomer comprising (a) a cap region which comprises a surface region that forms part of the upper surface of a pore formed from the modified actinobacterial pore monomer and (b) a constriction region that forms part of a constriction in a channel of a pore formed from the modified actinobacterial pore monomer, wherein the actinobacterial pore monomer is modified to increase the distance between the surface region and the constriction region.

[0009] The invention also provides:

[0010] - A construct comprising two or more covalently attached modified actinobacterial pore monomers of the invention.

[0011] - A pore comprising at least one modified actinobacterial pore monomer of the invention.

[0012] - A pore comprising at least one construct of the invention.

[0013] - An oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, wherein each modified actinobacterial pore monomer comprises (a) a cap region which comprises a surface region forming part of the upper surface of the oligomeric pore and (b) a constriction region forming part of a constriction in the channel, and wherein each actinobacterial pore monomer is modified to increase the distance between the upper surface of the oligomeric pore and the constriction.

[0014] - An oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, wherein each modified actinobacterial pore monomer comprises (a) a cap region which comprises a surface region forming part of the upper surface of the oligomeric pore, (b) a transmembrane region forming the channel and (c) a constriction region forming part of a constriction in the channel, and wherein the transmembrane region in each actinobacterial pore monomer is modified to increase the distance between the upper surface of the oligomeric pore and the constriction.

[0015] - An oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, wherein each modified actinobacterial pore monomer comprises (a) a cap region which comprises a surface region forming part of the upper surface of the oligomeric pore and (b) a constriction region forming part of a constriction in thechannel, and wherein the distance between the upper surface of the oligomeric pore and the constriction is at least 79 angstroms (Å).

[0016] - A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel and (2) an analyte binding protein, wherein the oligomeric pore comprises (a) a cap region which comprises an upper surface which interacts with the analyte binding protein and (b) a constriction in the channel, and wherein each actinobacterial pore monomer is modified to increase the distance between the upper surface and the constriction.

[0017] - A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel and (2) an analyte binding protein, wherein the oligomeric pore comprises (a) a cap region which comprises an upper surface which interacts with the analyte binding protein, (b) a transmembrane region forming the channel and (c) a constriction in the channel, and wherein the transmembrane region in each actinobacterial pore monomer is modified to increase the distance between the upper surface and the constriction.

[0018] - A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel and (2) an analyte binding protein, wherein the oligomeric pore comprises (a) a cap region which comprises an upper surface which interacts with the analyte binding protein and (b) a constriction in the channel, and wherein the distance between the upper surface and the constriction is at least 79 angstroms (Å).

[0019] - A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, (2) an analyte present in the channel of the pore, and (3) an analyte binding protein bound to the analyte, wherein the oligomeric pore comprises (a) a cap region which prevents the analyte binding protein from entering the channel and (b) a constriction in the channel, and wherein each actinobacterial pore monomer is modified to increase the minimum distance between the analyte binding protein and the constriction.

[0020] - A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, (2) an analyte present in the channel of the pore, and (3) an analyte binding protein bound to the analyte, wherein the oligomeric pore comprises (a) a cap region which prevents the analyte binding protein from entering the channel, (b) a transmembrane region forming the channel and (c) a constriction in the channel, and wherein the transmembrane region in each actinobacterial pore monomer is modified to increase the minimum distance between the analyte binding protein and the constriction.- A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, (2) an analyte present in the channel of the pore, and (3) an analyte binding protein bound to the analyte, wherein the oligomeric pore comprises (a) a cap region which prevents the analyte binding protein from entering the channel and (b) a constriction in the channel, and wherein the minimum distance between the analyte binding protein and the constriction is at least 79 angstroms (Å).

[0021] - A membrane comprising a pore of the invention, an oligomeric pore of the invention or a pore complex of the invention.

[0022] - A method for determining the presence, absence or one or more characteristics of a target analyte, comprising the steps of: (i) contacting the target analyte with a pore of the invention, an oligomeric pore of the invention or a pore complex of the invention, such that the target analyte moves with respect to the pore, oligomeric pore or pore complex; and (ii) taking one or more measurements as the analyte moves with respect to the pore, oligomeric pore or pore complex and thereby determining the presence, absence or one or more characteristics of the analyte. Use of a pore of the invention, an oligomeric pore of the invention or a pore complex of the invention to determine the presence, absence or one or more characteristics of a target analyte.

[0023] - A polynucleotide which encodes a modified actinobacterial pore monomer of the invention or a construct of the invention.

[0024] - A kit for characterising a target analyte comprising (a) a pore of the invention, an oligomeric pore of the invention or a pore complex of the invention and (b) the components of a membrane.

[0025] - A kit for characterising a target polynucleotide or a target polypeptide comprising (a) a pore of the invention or an oligomeric pore of the invention and (b) an analyte binding protein.

[0026] - An apparatus for characterising a target analyte in a sample, comprising (a) a plurality of pores of the invention or a plurality of oligomeric pores of the invention and (b) a plurality of analyte binding proteins.

[0027] - An array comprising a plurality of membranes of the invention.A system comprising (a) a membrane of the invention or an of the invention, (b) means for applying a potential across the membrane(s) and (c) means for detecting electrical or optical signals across the membrane(s).

[0028] - An apparatus comprising a pore of the invention, an oligomeric pore of the invention or a pore complex of the invention inserted into an in vitro membrane.

[0029] - An apparatus produced by a method comprising (i) obtaining a pore of the invention, an oligomeric pore of the invention or a pore complex of the invention and (ii) contacting the pore, pore complex, oligomeric pore or oligomeric pore complex with an in vitro membrane such that the pore, pore complex, oligomeric pore or oligomeric pore complex is inserted in the in vitro membrane.

[0030] - A method of improving the ability of an oligomeric pore formed from actinobacterial pore monomers surrounding a channel to characterise an analyte, wherein each actinobacterial pore monomer comprises (a) a cap region which comprises a surface region forming part of the upper surface of the oligomeric pore and (b) a constriction region forming a constriction in the channel, the method comprising modifying at least one of the actinobacterial pore monomers to increase the distance between the upper surface and the constriction.

[0031] - A method of improving the ability of a pore complex comprising (1) an oligomeric pore formed from actinobacterial pore monomers surrounding a channel and (2) an analyte binding protein to characterise an analyte, wherein each actinobacterial pore monomer comprises (a) a cap region which comprises an upper surface which interacts with the analyte binding protein and (b) a constriction in the channel, the method comprising modifying at least one of the actinobacterial pore monomers to increase the distance between the upper surface and the constriction.

[0032] - A method of improving the ability of a pore complex comprising (1) an oligomeric pore formed from actinobacterial pore monomers surrounding a channel, (2) an analyte present in the channel of the pore, and (3) an analyte binding protein bound to the analyte, wherein the oligomeric pore comprises (a) a cap region which prevents the analyte binding protein from entering the channel and (b) a constriction in the channel, the method comprising modifying at least one of the actinobacterial pore monomers to increase the distance between the minimum analyte binding protein and the constriction.

[0033] - A modified actinobacterial pore monomer having a root mean square deviation (RMSD) of less than about 4.0 Å when compared with a protein having the sequence shown in any one of SEQ ID NOs: 13-21.DESCRIPTION OF THE FIGURES

[0034] Figure 1: Structures of the MspA wild type and 2aa, 4aa, 6aa and 8aa pores and monomers. Predicted 3-dimensional pore structures for the elongated pores prepared according to the AlphaFold algorithm (Hassabis D et al, Nature, 596, 583-589, 2021).

[0035] Distance between the pore upper surface to the constriction shown for all pores.

[0036] Figure 2: Measurements of the MspA wild type pore structure (protein databank accession code 1UUN). For all measurements, the distances are between C-alpha to C-alpha atoms. The MspA pore consists of the cap region (A) and the transmembrane beta barrel region (B) which together make the cap region (or scaffold) (C), and the constriction region (D).

[0037] Figure 3: A schematic diagram of the DNA static strand experiment. Pores are probed with oligonucleotides (SS29-SS38) with an abasic nucleotide (asterisk) at a defined distance from a biotin (B)-streptavidin (S). Composition of the SS29 to SS38 strands are shown in Table 6, each featuring 3'Biotin and a iSpC3 abasic within a polyA nucleotide background.

[0038] Figure 4: DNA static strand experiment data shown for MspA non-elongated and the 2aa, 4aa, 6aa and 8aa elongated pore structures. Pores are sequentially probed with oligonucleotides, SS29-SS38. When the abasic residue resides at the pore constriction, an increase in conductance level is observed. Each dot represents a single data point.

[0039] Figure 5: DNA static strand experiment data shown for Mrh non-elongate and the 2aa, 4aa and 8aa elongated pore structures. Pores are sequentially probed with oligonucleotides, SS29-SS38. When the abasic residue resides at the pore constriction, an increase in conductance level is observed. Each dot represents a single data point.

[0040] Figure 6: Structure and dimensions of PorARc (cryoEM structure). This represents a PorARc before it is modified in accordance with the invention. The distances are measured from backbone to backbone of the amino acids forming the pore structure. The PorARc pore is a symmetrical octameric pore. The overall height is 90.4 A, and the largest outer dimension is 90.7 A. The PorARc pore consists of the cap region (A) and the transmembrane beta barrel region (B) which together make the cap region (or scaffold) (C), and the constriction region (D). The cap region (or scaffold) has a height of 73.6 A corresponding to height of the cap region (A), 44.7 A, and the transmembrane beta barrel region (B), 26.2 A. The constriction region (C) has a height of 19.7 A. The PorARc pore has an overall funnel shape with an entry width of 49 A which narrows to 41.5 A at the bottom of the cap region (A) and narrows further to 39.8 A at the transmembrane beta barrel region (B). The constriction region (C) has a sharp narrowing to 27.4 A and then widens to 36.1 A at the base of the pore structure.Figure 7: A schematic of the native and elongated pores with a motor-loaded peptide-DNA conjugate. The peptide is shown in black, and the DNA is in grey lines. The wider section in the peptide-DNA conjugate indicates the position of the asparagine, N, at position 15 in the peptide. Also shown in Figure 7 are example ionic current traces observed when the PolyE and PolyE_15N strands translocate through the nanopore.

[0041] Figure 8: A schematic of the native and elongated pores with a motor-loaded peptide-DNA conjugate. The peptide is shown in black, and the DNA is in grey lines. The wider section in the peptide-DNA conjugate indicates the position of the asparagine, N, at position 15 in the peptide. Also shown in Figure 8 are overlays of multiple ionic current traces as strands of the same species translocate through the nanopore.

[0042] DESCRIPTION OF THE SEQUENCE LISTING SEQ ID NO: 1 shows the amino acid sequence of PorARc from Rhodococcus corynebacteroides (PorARc_Rco) with the substitutions E78R / D82S / E116T / E125A / D165S.

[0043] SEQ ID NO: 2 shows the amino acid sequence of PorARc pore from Mycolicibacterium phlei (PorARc_Mph) with the substitutions D91N / D92N.

[0044] SEQ ID NO: 3 shows the amino acid sequence of PorARc pore from Mycobacterium sp. (PorARc_Msp) with the substitutions D91N / D92N.

[0045] SEQ ID NO: 4 shows the amino acid sequence of PorARc pore from Mycolicibacterium rhodesiae (PorARc_Mrh) with the substitutions D91N / D92N.

[0046] SEQ ID NO: 5 shows the amino acid sequence of PorARc pore from Mycolicibacterium elephantis (PorARc_Mel) with the substitutions D91N / E101Q.

[0047] SEQ ID NO: 6 shows the amino acid sequence of PorARc pore from Mycolicibacterium cosmeticum (PorARc_Mco) with the substitutions D91N / D92N.

[0048] SEQ ID NO: 7 shows the amino acid sequence of PorARc pore from unclassified Rhodococcus (WP_056447532.1; PorARc_Rsp) with the substitutions E89Q / D91N / D93N / D100N.

[0049] SEQ ID NO: 8 shows the amino acid sequence of PorARc pore from Rhodococcus sp PSBB049 (WP_206003768.1; PorARc_Rsp) with the substitutions D90N / D95N / D103N.

[0050] SEQ ID NO: 9 shows the amino acid sequence of PorARr.

[0051] SEQ ID NO: 10 shows the amino acid sequence of PorBRr.SEQ ID NO: 11 shows the amino acid sequence of wild-type MspA from Mycobacterium smegmatis.

[0052] SEQ ID NO: 12 shows the amino acid sequence of MspA_m2.

[0053] SEQ ID NOs: 13-33 shows the sequences used in the Examples.

[0054] DETAILED DESCRIPTION

[0055] All publications, patents and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety. All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent publications and patents or patent applications incorporated by reference contradict the invention contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material.

[0056] The present invention will be described with respect to particular embodiments and with reference to certain drawings but the invention is not limited thereto but only by the claims. Any reference signs in the claims shall not be construed as limiting the scope. Of course, it is to be understood that not necessarily all aspects or advantages may be achieved in accordance with any particular embodiment of the invention. Thus, for example those skilled in the art will recognize that the invention may be embodied or carried out in a manner that achieves or optimizes one advantage or group of advantages as taught herein without necessarily achieving other aspects or advantages as may be taught or suggested herein.

[0057] In addition, as used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "a pore monomer" includes two or more pore monomers, reference to "a constriction" includes two or more constrictions, reference to "a pore" includes two or more pores, reference to "an oligomeric pore" includes two or more oligomeric pores, reference to "a pore complex" includes two or more pore complexes, reference to "a target analyte" includes two or more target analytes, reference to "an analyte binding protein" includes two or more such proteins, reference to "a

[0058] helicase" includes two or more helicases, and the like.

[0059] In all instances here, the terms "comprises" or "comprising" cover and can be replaced by "consists of" or "consisting of".In all of the discussion herein, the standard one letter codes for amino acids are used. These are as follows: alanine (A), arginine (R), asparagine (N), aspartic acid (D), cysteine (C), glutamic acid (E), glutamine (Q), glycine (G), histidine (H), isoleucine (I), leucine (L), lysine (K), methionine (M), phenylalanine (F), proline (P), serine (S), threonine (T), tryptophan (W), tyrosine (Y) and valine (V). Standard substitution notation is also used, i.e., E89R means that E at position 89 is replaced with R.

[0060] In the paragraphs herein where different amino acids at a specific position are separated by the I symbol, the I symbol means "or". For instance, D90N / Q means D90N or D90Q. In the paragraphs herein where different positions or different substitutions are separated by the I symbol, the I symbol means "and" such that I87 / D90 is 187 and D90 and I87Q / D90S is I87Q and D90S.

[0061] A "pore" in the context of the invention is a transmembrane protein structure defining a channel or hole that allows the translocation of molecules and ions from one side of the membrane to the other. The translocation of ionic species through the pore may be driven by an electrical potential difference applied to either side of the pore. A "nanopore" is a biological pore in which the minimum diameter of the channel through which molecules or ions pass is in the order of nanometres (10-9nanometres). In some embodiments, the pore can be a transmembrane protein pore. The transmembrane protein structure of a biological pore may be monomeric or oligomeric in nature. Typically, the pore comprises a plurality of polypeptide monomers or subunits arranged around a central axis thereby forming a protein-lined channel that extends substantially perpendicular to the membrane in which the pore resides. The number of polypeptide monomers or subunits is not limited. Typically, the number of monomers or subunits is from 5 to up to 30, suitably the number of monomers or subunits is from 6 to 10. The portions of the protein monomers or subunits within the pore that form protein-lined channel typically comprise secondary structural motifs that may include one or more trans-membrane [3-barrel, and / or o-helix sections.

[0062] In its broadest context, the term "pore" covers a pore complex or a pore multimer. A "pore complex" of the invention typically comprises pore monomers or an oligomeric pore and an analyte binding protein

[0063] " About" as used herein when referring to a measurable value such as a percentage or an amount and the like, is meant to encompass variations of ± 20 % or ± 10 %, more preferably ± 5 %, even more preferably ± 1 %, and still more preferably ± 0.1 % from the specified value, as such variations are appropriate to perform the disclosed methods. Any statement herein including the term "about" includes the same feature without the term. For instance, at least "about" 20% includes at least 20%.Corresponding positions or regions may be determined by standard techniques in the art. For example, the PILEUP and BLAST algorithms mentioned below can be used to align the sequence of the invention, such as a pore monomer, with the reference sequence, e.g., SEQ ID NO: 1, and identify corresponding positions or regions.

[0064] Standard methods in the art may be used to determine homology and / or identity. For example, the UWGCG Package provides the BESTFIT program which can be used to calculate homology and identity, for example used on its default settings (Devereux et al (1984) Nucleic Acids Research 12, p387-395). The PILEUP and BLAST algorithms can be used to calculate homology or line up sequences (such as identifying equivalent residues or corresponding sequences (typically on their default settings)), for example as described in Altschul S. F. (1993) J Mol Evol 36:290-300; Altschul, S. F et al (1990) J Mol Biol 215:403-10. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov / ).

[0065] Standard methods in the art can be used to determine structural similarity, including RMSD. Suitable methods include, but are not limited to, AlphaFold, PSIPRED, TM-Align, US-Align or FATCAT. In all instances herein, RMSD is preferably measured over all alpha carbon atoms (Cα atoms). RMSD may be measured over all heavy atoms.

[0066] Various embodiments below are defined with reference to a "part" of another feature. For example, the surface region may be part of an external surface of the cap region or the constriction region forms part of a constriction in a channel. In all embodiments herein, the part may be any amount of the reference embodiment, such as the cap region, upper surface or constriction. The part be at least about 5% of the reference embodiment, such as the cap region, upper surface or constriction. The part may be at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 98% or at least about 99% of the reference embodiment, such as the cap region, upper surface or constriction. The part may be 100% of the reference embodiment, such as the cap region, upper surface or constriction. %s are usually calculated on the basis on the number of amino acids in the part compared with the number of amino acids in reference embodiment. The part may comprise at least about 3 amino acids, such as at least about 3 amino acids, at least about 4 amino acids, at least about 5 amino acids, at least about 10 amino acids, at least about 15 amino acids, at least about 20 amino acids, at least about 30 amino acids, at least about 40 amino acids, at least about 50 amino acids, at least about 60 amino acids, at least about 70 amino acids, at least about 80 amino acids, at least about 90 amino acids, at least about 100 amino acids, at least about 110 amino acids, at least about 120 amino acids, at least about 130 amino acids, at least about 140 amino acids or at least about 150 amino acids.The general definitions in WO 2019 / 002893 are incorporated by reference herein in their entirety.

[0067] Pore monomers

[0068] The invention provides a modified pore monomer. The term "pore monomer" is a term of the art. Pore monomers are known in the art and numerous examples are discussed in more detail below. The modified pore monomer is typically capable of forming a pore comprising a channel. The ability of the modified pore monomer to form a pore comprising a channel can be measured using routine methods, including any of those described in WO 2016 / 034591, WO 2017 / 149316, WO 2017 / 149317, WO 2017 / 149318, WO 2018 / 211241, WO 2019 / 002893, WO 2023 / 118404, WO 2023 / 198911, WO 2024 / 033421, WO 2024 / 033422, WO 2024 / 033443, WO 2024 / 089270 (all incorporated by reference herein in their entirety) and in the Example. The ability of the modified pore monomer to form a pore comprising a channel can be measured using the methods described in any of the patent applications cited below.

[0069] In all of the discussion below, the term "modified" means the pore monomer of the invention is not a wild-type or native pore monomer. The pore monomer of the invention is modified because the distance between its surface region and its constriction region has been increased. The distance may be increased in any of the ways discussed below. The skilled person may determine whether or not a pore monomer has been modified or elongated in accordance with the invention by measuring the distance between the surface region and the constriction region and comparing it with a wild-type, native or natural pore monomer.

[0070] In all of the discussion above and below, pore monomers before they are modified in accordance with the invention ( / .e., pore monomers used to construct the modified pore monomers of the invention) are referred to as a / the pore monomer or pore monomers ( / .e., without the term "modified"). Pore monomers modified in accordance with the invention of course include the term "modified", i.e., a / the modified pore monomer or modified pore monomers.

[0071] The invention provides a modified pore monomer comprising (a) a cap region which comprises a surface region that forms part of the upper surface of a pore formed from the modified pore monomer and (b) a constriction region that forms a constriction in a channel of a pore formed from the modified pore monomer, wherein the pore monomer is modified to increase the distance between the surface region and the constriction region.

[0072] Specific regions, such as the cap region or constriction region, in a pore may be identified using standard methods. Regions in proteins of unknown structure can be defined by aligning the protein sequence with a homologous protein of known structure. If there issufficiently high sequence identity or similarity between the sequences, there is confidence in the region boundaries. It is possible to add further confidence in a region boundary through use of tools that predict tertiary structure (e.g., AlphaFold) or secondary structural elements (e.g., PSIPRED). For the latter, if both the protein of unknown structure and the protein of known structure are flanked by the same predicted secondary structural element, then there is more confidence of the region boundary.

[0073] Cap regions are known in the art and are discussed in more detail below. The cap region typically forms the structural core of the protein pore and may partially reside in a membrane. When the cap region forms the structural core and partially resides in a membrane, it may also be known as a scaffold. For instance, a cap region and a transmembrane region together may be known as a scaffold. The cap region of the modified pore monomer typically forms part of a cap region in a pore formed from the modified pore monomer. The cap region "forms part" of the cap region in the pore in the sense it forms the cap region with cap regions from the other pore monomers, including modified pore monomers, used to form the pore. As explained in more detail below, the modified pore monomers of the invention may be used to form the pores, oligomeric pores and pore complexes of the invention.

[0074] The cap region (or scaffold) is preferably from about 67 to about 187 amino acids in length. The cap region (or scaffold) is preferably about 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, or 187 amino acids in length.

[0075] The cap region (or scaffold) is preferably from about 87 to about 167 amino acids in length. The cap region (or scaffold) is preferably about 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167 amino acids in length. The cap region (or scaffold) is preferably about 87, 159, 160, 166, or 167 amino acids in length. The cap region may have may any of the dimensions discussed below.The cap region in the modified pore monomer comprises a surface region. The surface region is part or all of an external surface of the cap region. Parts are defined above. The skilled person is capable of identifying an external surface of the cap region. The three-dimensional structure of a modified pore monomer and / or its cap region can be determined using standard methods in the art (e.g., AlphaFold or PSIPRED) and the skilled person can therefore identify the external surface of the cap region. The surface region typically comprises one or more of (a) one or more loop regions, (b) one or more alpha helices and (c) one or more beta strands or sheets, such as (a), (b), (c), (a) and (b), (b) and (c), (a) and (c) or (a), (b) or (c). The surface region typically comprises one or more loop regions and one or more beta strands or sheets.

[0076] The surface region forms part of the upper surface of a pore formed from the modified pore monomer. The surface region "forms part" of the upper region surface in the sense it forms the upper surface with surface regions from the other pore monomers, including other modified pore monomers, used to form the pore. As explained in more detail below, the modified pore monomers of the invention may be used to form the pores, oligomeric pores and pore complexes of the invention. The upper surface of the pore is usually the upper external surface of the pore when viewed from the side. An example of a side view is shown in Figures 2 and 6 The upper surface may be any size and shape. The upper surface is usually ring-shaped when viewed from the top. The ring-shaped upper surface typically surrounds the upper opening of the channel. The three-dimensional structure of a pore can be determined using standard methods in the art (e.g., AlphaFold or PSIPRED) and the skilled person can therefore identify the upper surface. The upper surface typically comprises one or more of (a) one or more loop regions, (b) one or more alpha helices and (c) one or more beta strands or sheets, such as (a), (b), (c), (a) and (b), (b) and (c), (a) and (c) or (a), (b) or (c). The surface region typically comprises one or more loop regions and one or more beta strands or sheets.

[0077] The upper surface typically interacts with, such as physically or sterically interacts with, an analyte binding protein. The upper surface typically prevents, such as physically or sterically prevents, an analyte binding protein from entering the channel of the pore. The upper surface is typically capable of interacting with, such as physically or sterically interacting with, an analyte binding protein. The upper surface is typically capable of preventing, such as physically or sterically preventing, an analyte binding protein from entering the channel of the pore. The modified pore monomer may be used to form pores for characterising a target analyte as discussed in more detail below. The movement of the target analyte with respect to the pore, such as through the pore, may be controlled using an analyte binding protein, which is too large to enter and move through the channel of the pore. When the target analyte and the analyte binding protein are contacted with the top of the pore, the target analyte moves through the channel of the pore but the analyte binding protein isprevented from entering the channel by the upper surface due to a physical or steric interaction.

[0078] Constriction regions are known in the art and are discussed in more detail below. The constriction region "forms part" of the constriction in the pore in the sense it forms the constriction with constriction regions from the other pore monomers, including other modified pore monomers, used to form the pore. As explained in more detail below, the modified pore monomers of the invention may be used to form the pores, oligomeric pores and pore complexes of the invention. The constriction region is typically capable of forming a channel in a pore formed from the modified pore monomer. Methods for measuring the ability to form a pore are discussed above.

[0079] The modified pore monomer may comprise any number of constriction regions. The modified pore monomer may comprise two or more constriction regions. The modified pore monomer may comprise any number of two or more constriction regions, such as 2, 3, 4, 5, 6 or more. The modified pore monomer may comprise a first constriction region which forms part of a first constriction in the channel, a second constriction region which forms part of a second constriction in the channel and a third constriction region which forms part of a third constriction in the channel.

[0080] The term "constriction", "orifice", "channel constriction", or "constriction site", as used interchangeably herein, refers to an aperture defined by a luminal surface of a pore, which acts to allow the passage of ions and target analytes through the pore channel.

[0081] Constrictions and constriction regions are known in the art. The constriction formed by the constriction regions are typically the narrowest apertures within a pore or within the channel defined by the pore. The constriction may be defined functionally. For instance, the constriction may be identified using the level of discrimination of a target analyte, such as a target polynucleotide or target polypeptide. Peak discrimination levels may identify the constriction. Discrimination is the contribution of the constriction to overall changes in ionic current level when target analyte, such as a DNA molecule, is translocated through the pore. A skilled person is capable of doing this, for instance as described in the Example.

[0082] The constriction may serve to limit the passage of molecules through the pore. The size of the constriction is typically a key factor in determining suitability of a pore for target analyte characterisation. If the constriction is too small, the target analyte to be characterised will not be able to pass through. However, to achieve a maximal effect on ion flow through the channel, the constriction should not be too large. For example, the constriction should not be wider than the solvent-accessible transverse diameter of a target analyte. Ideally, any constriction should be as close as possible in diameter to the transverse diameter of the analyte passing through.The narrowing of a central cavity or aperture typically forms a constriction in the channel. In some embodiments, the diameter of a constriction is calculated by measuring the distance between the alpha-carbons (Cα) of the amino acid residues that extend furthest into the channel of the pore to form the constriction. In some embodiments, the diameter of a constriction is calculated by measuring the distance between the Van der Waals radii of the atoms extending furthest into the channel of the pore to form the constriction. In some embodiments, the minimum diameter of a constriction ranges from about 0.5 nanometres (nm) to about 4.0 nm (e.g., as measured by the distance between Van der Waals radii). In some embodiments, the minimum diameter of the constriction ranges from about 0.5 to about 3.0 nm or from about 0.5 to about 2.0 nm, preferably from about 0.7 to about 1.8 nm, from about 0.8 to about 1.7 nm, from about 0.9 to about 1.6 nm, or from about 1.0 to about 1.5 nm, such as about 1.1, about 1.2, about 1.3 or about 1.4 nm. In some embodiments, the minimum diameter of a constriction ranges from about 10 A to about 30 A, for example about 10 A, about 11 A, about 12 A, about 13 A, about 14 A, about 15 A, about 16 A, about 17 A, about 18 A, about 19 A, about 19.2 A, about 20 A, about 21 A, about 22 A, about 23 A, about 24 A, about 25 A, about 26 A, about 27 A, about 28 A, about 29 A, or about 30 A (e.g., as measured by Cα to Cα). In some embodiments, the minimum diameter of a constriction ranges from about 10 A to about 30 A (e.g., as measured by Cα to Cα). In some embodiments, the minimum diameter of a constriction ranges from about 15 A to about 25 A (e.g., as measured by Cα to Cα).

[0083] The constriction region is preferably from about 5 to about 114 amino acids in length. The constriction region is preferably about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, about 50, about 51, about 52, about 53, about 54, about 55, about 56, about 57, about 58, about 59, about 60, about 61, about 62, about 63, about 64, about 65, about 66, about 67, about 68, about 69, about 70, about 71, about 72, about 73, about 74, about 75, about 76, about 77, about 78, about 79, about 80, about 81, about 82, about 83, about 84, about 85, about 86, about 87, about 88, about 89, about 90, about 91, about 92, about 93, about 94, about 95, about 96, about 97, about 98, about 99, about 100, about 101, about 102, about 103, about 104, about 105, about 106, about 107, about 108, about 109, about 110, about 111, about 112, about 113, or about 114 amino acids in length.

[0084] The constriction region is preferably from about 15 to about 94 amino acids in length. The constriction region is preferably about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, about 50, about 51, about 52, about 53, about 54, about 55, about 56, about 57, about 58, about 59, about 60, about 61, about 62, about 63, about 64, about 65, about 66, about 67, about 68, about 69, about 70, about 71, about 72, about 73, about 74, about 75, about 76, about 77, about 78, about 79, about 80, about 81, about 82, about 83, about 84, about 85, about 86, about 87, about 88, about 89, about 90, about 91, about 92, about 93, about or 94 amino acids in length. The constriction region is preferably about 15, about 16, about 17, about 18, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 39, about 50, about 52, about 86, about 94 amino acids in length. Additional constriction regions may be any of these lengths.

[0085] The constriction region is preferably from about 6 to about 46 amino acids in length. The constriction region is preferably about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45 or 46 amino acids in length. The constriction region is preferably from about 16 to about 36 amino acids in length. The constriction region is preferably about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35 or 36 amino acids in length. The constriction region is preferably about 26 amino acids in length.

[0086] The constriction region may have may any of the dimensions discussed below.

[0087] In all instances herein, the term "distance" may comprise one or more of (a) the vertical distance between the surface region and the constriction region, (b) the number of amino acids between the surface region and the constriction region and (c) the equivalent number of nucleotides between the surface region and the constriction region. The term "distance" may comprise (a), (b), (c), (a) and (b), (a) and (c), (b) and (c), or (a), (b) and (c). The vertical distance is typically measured when the modified pore monomer, pore, oligomeric pore or pore complex is viewed from the side (see, e.g., the side view in Figure 2). The vertical distance is typically measured in Angstroms (A). The number of amino acids between the surface region and the constriction region may be counted in a straightforward manner. The equivalent number of nucleotides between the surface region and the constriction region can be measured when a polynucleotide analyte passes through a pore, an oligomeric pore or pore complex formed from the modified pore monomer.Before modification in accordance with the invention, the distance, such as vertical distance, between the surface region and the constriction region may be from about 5 A to about 78 A, such as from about 10 A to about 77 A, from about 15 A to about 76 A, from about 20 A to about 75 A or from about 25 A to about 70 A. This can be measured as the distance between the alpha-carbons (Cα) of the amino acid residue extending furthest upwards in the upper surface and the amino acid residue in the constriction region extending furthest into the lumen of the channel.

[0088] Before modification in accordance with the invention, the distance, such as vertical distance, between the surface region and the constriction region is about 5 A, about 6 A, about 7 A, about 8 A, about 9 A, about 10 A, about 11 A, about 12 A, about 13 A, about 14 A, about 15 A, about 16 A, about 17 A, about 18 A, about 19 A, about 20 A, about 21 A, about 22 A, about 23 A, about 24 A, about 24.8 A, about 25 A, about 26 A, about 27 A, about 28 A, about 29 A, about 30 A, about 31 A, about 32 A, about 33 A, about 34 A, about 35 A, about 36 A, about 37 A, about 38 A, about 39 A, about 40 A, about 41 A, about 42 A, about 43 A, about 44 A, about 45 A, about 46 A, about 47 A, about 48 A, about 49 A, about 50 A, about 51 A, about 52 A, about 53 A, about 54 A, or about 55 A. In some embodiments, the distance between the first constriction region and the second constriction region is more than about 55 A in length, such as about 56 A, about 57 A, about 58 A, about 59 A, about 60 A, about 61 A, about 62 A, about 63 A, about 64 A, about 65 A, about 66 A, about 67 A, about 68 A, about 69 A, about 70 A, about 71 A, about 72 A, about 73 A, about 74 A, about 75 A, about 76 A, about 77 A, or about 78 A in length. Before modification in accordance with the invention, the distance between the upper surface and the constriction region is less than 78 A in length.

[0089] The pore monomer typically further comprises a transmembrane region. This region is typically capable of forming a channel. This can be measured as discussed above. The modified pore monomer may comprise a cap region, a transmembrane region and a constriction region.

[0090] The transmembrane region is preferably from about 28 to about 68 amino acids in length. The transmembrane region is preferably about 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67 or 68 amino acids in length. The transmembrane region is preferably from about 15 to about 30 amino acids in length. The transmembrane region is preferably about 15, 16, 17 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 amino acids in length.

[0091] The transmembrane region may have any of the dimensions and / or lengths discussed below. The transmembrane region may have any of the dimensions and / or lengths described in WO 2024 / 089270 (incorporated by reference herein in its entirety).The transmembrane region may be a transmembrane beta barrel region or a transmembrane alpha helical region. The transmembrane region is preferably a transmembrane beta barrel region.

[0092] The pore monomer used to construct the modified pore monomer may be from or derived from any pore. The modified pore monomer may be from or derived from any pore. The pore monomer or modified pore monomer may be from or derived from a bacterial pore. The pore monomer or modified pore monomer may be from or derived from a terrabacterial pore. As discussed below, the pore monomer or modified pore monomer is preferably from or derived from an actinobacterial pore.

[0093] The pore monomer or modified pore monomer is preferably from or derived from Wza, Iota toxin, Anthrax protective antigen, Vibrio cholerae cytolysin, Cytotoxin K (CytK), CELIII, CsgG, CsgF, CsgG-CsgF, Aerolysin, alpha hemolysin, MspA, MspB, MspC, PorARr, PorBRr, PorARc, PilQ, necrotic enteritis B-like toxin (NetB), FraC, portal proteins including G20c, P23_45, T4, SPP1, P22 and Phi29, gamma hemolysin, Monalysin, Lysenin, ClyA, an actinoporin, Clostridium perfringens beta toxin, parasporin-2, epsilon toxin, lectin from the parasitic mushroom Laetiporus sulphureus (LSL), volvatoxin, Cry toxins, CytlAa, Cyt2Aa, Complement component 9 (C9), Perfringolysin O, Pleurotolysin, Listeriolysin, Perforin-2, Gasdermin-A3, L-, P- and M-ring protein, Type II secretion system protein D, GspD, InvG, VirB7, SpoIIIAG, Cag8, Cag3, Cag or other proteins in the Type IV secretion system apparatus protein CagY, WzzB, Pentraxin, Afp2, Major vault protein, Thioredoxin-dependent peroxidase reductase, Arf-GAP, Respiratory syncytial virus ribonucleoprotein, Chikungunya virus nonstructural protein 1, PRC, YaxA, XaxA, HfaB, NfpAB, leucocidin, PrgH, TraT, TolC, TamA, SprA or Sov. Suitable actinoporins include, but are not limited to, actinoporins from or derived from Orbicella faveolata, such as those described in PCT / EP2024 / 060202.

[0094] The pore monomer or modified pore monomer is preferably from or derived from Iota toxin, Anthrax protective antigen, Vibrio cholerae cytolysin, Cytotoxin K (CytK), CELIII, CsgG, CsgG-CsgF, Aerolysin, alpha hemolysin, MspA, MspB, MspC, PorARr, PorBRr, PorARc, PilQ, necrotic enteritis B-like toxin (NetB), gamma hemolysin, Monalysin, Lysenin, Clostridium perfringens beta toxin, parasporin-2, epsilon toxin, lectin from the parasitic mushroom Laetiporus sulphureus (LSL), Cry toxins, CytlAa, Cyt2Aa, Complement component 9 (C9), Perfringolysin O, Pleurotolysin, Listeriolysin, Perforin-2, Gasdermin-A3, L-, P- and M-ring protein, Type II secretion system protein D, GspD, InvG, SpoIIIAG, YaxA, XaxA, HfaB, NfpAB, leucocidin, TraT, TolC, TamA, SprA or Sov. The pore monomer or modified pore monomer may be from or derived from any of the pores described in WO 2016 / 034591, WO 2017 / 149316, WO 2017 / 149317, WO 2017 / 149318, WO 2018 / 211241, WO 2019 / 002893, WO 2023 / 118404, WO 2023 / 198911, WO 2024 / 033421, WO 2024 / 033422, WO 2024 / 033443, and WO 2024 / 089270 (all incorporated by reference herein in their entirety).The pore monomer or modified pore monomer may be from or derived from any of the MspA pores described in WO 2012 / 107778, WO 2015 / 166275, WO 2016 / 055778, WO 2016 / 132124, and WO 2016 / 132123 (all incorporated herein by reference in their entireties) or a variant thereof.

[0095] The pore monomer or modified pore monomer may be from or derived from any of the CsgG pores described in WO 2023 / 060420, WO 2023 / 60418, WO 2023 / 60422, WO 2023 / 060421, WO 2023 / 019470, CN114957412, WO 2023 / 019471, W02023 / 060419 and WO 2023 / 050031 (all incorporated herein by reference in their entireties) or a variant thereof.

[0096] The pore monomer or modified pore monomer may be from or derived from any of the pores described in WO 2023 / 123370, WO 2024 / 138470, WO 2024 / 138472, WO 2024 / 138424, WO 2024 / 138425, WO 2024 / 138512 and WO 2024 / 138565 (all incorporated herein by reference in their entireties) or a variant thereof.

[0097] A pore monomer is "from" or "derived from" a pore if it shares structural similarity with a monomer in the wild-type or native pore. The pore monomer may share structural similarity before modification in accordance with the invention. The modified pore monomer may share structural similarity after modification in accordance with the invention. The pore monomer or modified pore monomer is preferably a structural variant having a root mean square deviation (RMSD) of less than about 4.0 Angstroms (A), such as less than about 3.5 A, less than about 3.0 A, less than about 2.5 A, less than about 2.0 A, less than about 1.5 A, less than about 1.0 A or less than about 0.5 A, when compared with the wild-type or native monomer.

[0098] Structural homologues typically have similar sequences. The pore monomer or modified pore monomer may be "from" or "derived from" a pore if it shares significant homology and / or identity with a monomer from the wild-type or native pore. The pore monomer or modified pore monomer preferably comprises a variant sequence having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity to the sequence of an wild-type or native monomer. The pore monomer or modified pore monomer preferably comprises a variant sequence having at least about 80% homology and / or identity to the sequence of an wild-type or native monomer. The pore monomer or modified pore monomer preferably comprises a sequence having at least about 90% homology and / or identity to the sequence of the wild-type or native monomer. The pore monomer preferably comprises a sequence having 100% homology and / or identity to the sequence of the wild-type or nativemonomer. Homology and / or identity is typically measured over the entire length of the monomer.

[0099] Sequence homology and / or identity can also relate to a fragment or portion of the reference sequence. Hence, a sequence may have less than 40% overall sequence homology and / or identity with the wild-type or native monomer, but the sequence of a particular region, domain or subunit could share at least about 80%, at least about 90%, or as much as about 99% sequence homology and / or identity with the corresponding region of the wild-type or native monomer. There may be at least about 80%, at least 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity over a stretch of 100 or more, for example 125, 150, 175 or 200 or more, contiguous amino acids ("hard homology").

[0100] The pore monomer or modified pore monomer may be from or derived from a chimeric pore monomer comprising two or more regions, wherein at least two of the two or more regions are from at least two different pores. The terms "from" and "derived from" are defined above. The chimeric pore monomer may comprise any number of regions, such as three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more or ten or more regions, from different pores. The chimeric pore monomer may comprise two or three regions. The regions may be any of those discussed above. The regions are preferably selected from a cap region, a constriction region, and a transmembrane region. The regions may be a cap region and a constriction region. The regions may be a cap region, a constriction region, and a transmembrane region. The at least two different pores are typically at least two different pores that appear in nature. The at least two different pores are typically at least two different wild-type or naturally occurring pores. The at least two different pores are preferably different before any artificial or synthetic modifications, such as additions, deletions and / or substitutions, are made to them. The at least two different pores are preferably homologues, for example structural homologues. A structural homologue refers to a protein or molecule that shares a similar three-dimensional structure with another protein or molecule. This can be determined using standard methods in the art (e.g., AlphaFold or PSIPRED). Structural homologues typically have similar sequences. Structural homologues are normally identified in similar species. The at least two different pores may be selected from any of the pores listed above. The at least two different pores may be selected from MspA, MspB, MspC, MspD, PorARr, PorBRr and PorARc. The at least two different pores may be two different PorARc pores or three different PorARc pores. The at least two different pores may be two different CsgG pores or three different CsgG pores. The chimeric pore monomer may be any of those described in WO 2024 / 089270 (incorporated by reference herein in its entirety).The pore monomer or modified pore monomer is preferably not from or derived from CsgG. The pore monomer or modified pore monomer is preferably not from or derived from FhuA. The terms "from" and "derived from" are defined above.

[0101] Modified actinobacterial pore monomers

[0102] The pore monomer is preferably from or derived from an actinobacterial pore monomer. The modified pore monomer is preferably from or derived from an actinobacterial pore monomer. The terms "from" and "derived from" are defined above.

[0103] In discussion below, pore monomers from or derived from actinobacterial pores may be referred to as actinobacterial pore monomers. They be actinobacterial pore monomers before modification in accordance with the invention, e.g., an actinobacterial pore monomer. They may be actinobacterial pore monomers modified in accordance with the invention, e.g., a "modified" actinobacterial pore monomer.

[0104] In all of the discussion below relating to increasing a distance, the terms "pore monomer" and "modified pore monomer" preferably relate to "an actinobacterial pore monomer" or "a modified actinobacterial pore monomer" respectively. When it comes to actinobacterial pore monomers, the term "pore" and "porin" are interchangeable.

[0105] In all instances herein, the term "actinobacterial" is interchangeable with "actinomycetotal", "actinobacteraeotal", "actinobacteriotai" or "actinomycetal".

[0106] Actinobacteria are a diverse phylum of Gram-positive bacteria with high GC content.

[0107] Suitable actinobacteria and actinobacterial pores are known in the art and are discussed below. The actinobacterium may be any of those described in Table 1. The actinobacterium may be Mycobacterium smegmatis.

[0108] The invention provides modified actinobacterial pore monomer comprising (a) a cap region which comprises a surface region that forms part of the upper surface of a pore formed from the modified actinobacterial pore monomer and (b) a constriction region that forms part of a constriction in a channel of a pore formed from the modified actinobacterial pore monomer, wherein the actinobacterial pore monomer is modified to increase the distance between the surface region and the constriction region. All of the discussion above concerning cap regions, surface regions, upper surfaces, constriction regions, constrictions and channels equally apply to these modified pore monomers.

[0109] PorARc, PorARr or PorBRr

[0110] The pore monomer is preferably from or derived from PorARc, PorARr or PorBRr. The modified pore monomer is preferably from or derived from PorARc, PorARr or PorBRr. Theactinobacterial pore monomer or the modified actinobacterial pore monomer is preferably from or derived from PorARc, PorARr or PorBRr. The pore monomer is preferably from or derived from PorARc. The modified pore monomer is preferably from or derived from PorARc. The actinobacterial pore monomer or the modified actinobacterial pore monomer is preferably from or derived from PorARc.

[0111] The terms "from" and "derived from" are defined above. The pore monomer or the modified pore monomer may be, may be from or may be derived from any of the monomers described in WO 2023 / 118404 and WO 2024 / 089270 (incorporated by reference herein in their entireties).

[0112] In discussion below, pore monomers from or derived from PorARc, PorARr or PorBRr may be referred to as PorARc, PorARr or PorBRr pore monomers respectively. They may be PorARc, PorARr or PorBRr pore monomers before modification in accordance with the invention, e.g., a PorARc pore monomer. They may be PorARc, PorARr or PorBRr pore monomers modified in accordance with the invention, e.g., a "modified" PorARc pore monomer.

[0113] In all of the discussion below relating to increasing a distance, the terms "pore monomer" and "modified pore monomer" preferably relate to "a PorARc, PorARr or PorBRr pore monomer" or "a modified PorARc, PorARr or PorBRr pore monomer" respectively.

[0114] The PorARc, PorARr or PorBRr pore preferably comprises a cap region (or scaffold) (e.g., C in Figure 6) and a constriction region (e.g., D is Figure 6). The PorARc, PorARr or PorBRr pore preferably comprises one or more of (a) a cap region (e.g. A in Figure 6), (b) a constriction region (e.g., D in Figure 6), and (c) a transmembrane beta barrel region (e.g., B in Figure 6), such as (a), (b), (c), (a) and (b), (a) and (c), (b) and (c), or (a), (b) and (c). The cap region and the transmembrane region may together be known as a scaffold. The PorARc, PorARr or PorBRr pore preferably comprises (a)-(c).

[0115] A PorARc pore monomer is capable of forming a PorARc pore. Before modification in accordance with the invention, the PorARc pore may be any size but preferably has the dimensions of the wild-type PorARc_Rco (Figure 6). The PorARc pore preferably has an external diameter of from about 70 to about 110 Å at its widest point, such as from about 80 to about 100 Å or from about 85 to about 95 Å at its widest point. The PorARc pore preferably has an external diameter of about 90.7 Å at its widest point. The PorARc pore preferably has a total length of from about 70 to about 110 Å, such as from about 80 to about 100 Å or from about 85 to about 95 Å. The PorARc pore preferably has a total length of about 90.4 Å. References to "total length" and "length" relate to the length of the pore or pore region when viewed from the side (see, e.g., the side view in Figure 6).The cap region (A in Figure 6) preferably has a length of from about 25 to about 65 Å, such as from about 35 to about 55 Å or from about 40 to about 50 Å. The cap region preferably has a length of about 44.7 Å. The channel defined by the cap region preferably has an opening of from about 30 to about 70 Å in diameter, such as from about 40 to about 60 Å or from about 45 to about 55 Å in diameter. The channel defined by the cap region preferably has an opening of about 49 Å in diameter. The channel defined by the cap region is preferably from about 20 to about 60 Å in diameter at its narrowest point, such as from about 30 to about 50 Å or from about 35 to about 45 Å in diameter at its narrowest point. The channel defined by the cap region is preferably about 41.5 Å in diameter at its narrowest point.

[0116] The transmembrane beta barrel region (B in Figure 6) preferably has a length of from about 5 to about 45 Å, such as from about 15 to about 35 Å or from about 20 to about 30 Å. The transmembrane beta barrel preferably has a length of about 26.2 Å. The channel defined by the transmembrane beta barrel region is preferably from about 20 to about 60 Å in diameter at its narrowest point, such as from about 30 to about 50 Å or from about 35 to about 45 Å in diameter at its narrowest point. The channel defined by the transmembrane beta barrel region is preferably about 39.8 Å in diameter at its narrowest point.

[0117] The cap region (or scaffold) (C in Figure 6 and formed from A and B) preferably has a length of from about 55 to about 95 Å, such as from about 65 to about 85 Å or from about 70 to about 80 Å. The cap region (or scaffold) preferably has a length of about 73.6 Å. The channel defined by the cap region (or scaffold) (C) is preferably from about 20 to about 60 Å in diameter at its narrowest point, such as from about 30 to about 50 Å or from about 35 to about 45 Å in diameter at its narrowest point. The channel defined by the cap region (or scaffold) (C) is preferably about 39.8 Å in diameter at its narrowest point.

[0118] The constriction region (D in Figure 6) preferably has a length of from about 5 to about 40 Å, such as from about 10 to about 30 Å or from about 15 to about 25 Å. The constriction region preferably has a length of about 19.7 Å. The channel defined by the constriction region is preferably from about 10 to about 50 Å in diameter at its narrowest point, such as from about 20 to about 40 Å, from about 22 to about 32 Å or from about 25 to about 35 Å in diameter at its narrowest point. The channel defined by the constriction region is preferably about 27.4 Å in diameter at its narrowest point. The channel defined by the constriction region is preferably from about 10 to about 60 Å in diameter at its narrowest point, such as from about 15 to about 55 Å, from about 25 to about 45 Å or from about 30 to about 40 Å in diameter at the base of the pore structure. The channel defined by the constriction region is preferably about 36.1 Å in diameter at the base of the pore structure. The constriction region is preferably from about 20 to about 60 Å in diameter, such as fromabout 30 to about 50 Å or from about 35 to about 45 Å. The constriction region is preferably about 41.9 Å in diameter.

[0119] All of the measurements above are based on measuring from backbone to backbone of the amino acids forming the different regions (as shown in Figure 6). All of the measurements typically apply to the PorARc pore monomer or PorARc pore before it is modified in accordance with the invention.

[0120] The regions in PorARc may have any of the amino acid lengths described in WO 2024 / 089270 (incorporated by reference herein in its entirety). The PorARc pore may be any of those in WO 2023 / 118404 or Table 2 of WO 2024 / 089270 (incorporated by reference herein in their entireties).

[0121] The PorARc pore monomer or modified PorARc pore monomer preferably comprises a cap region (or scaffold) (e.g., C in Figure 6) and a constriction region (e.g., D is Figure 6). The PorARc pore monomer or modified PorARc pore monomer preferably comprises one or more of (a) a cap region (e.g. A in Figure 6), (b) a constriction region (e.g., D in Figure 6), and (c) a transmembrane beta barrel region (e.g., B in Figure 6), such as (a), (b), (c), (a) and (b), (a) and (c), (b) and (c), or (a), (b) and (c). The cap region and the transmembrane region may together be known as a scaffold. The PorARc pore monomer or modified PorARc pore monomer preferably comprises (a)-(c).

[0122] The cap region (or scaffold) (e.g., C in Figure 6) in the PorARc pore monomer or modified PorARc pore monomer is preferably from about 67 to about 187 amino acids in length. The cap region (or scaffold) in the PorARc pore monomer or modified PorARc pore monomer is preferably about 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, or 187 amino acids in length. The cap region (or scaffold) in the PorARc pore monomer or modified PorARc pore monomer is preferably from about 87 to about 167 amino acids in length. The cap region (or scaffold) the PorARc pore monomer or modified PorARc pore monomer is preferably about 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167 amino acids in length. The cap region (or scaffold)in the PorARc pore monomer or modified PorARc pore monomer is preferably about 87, 159, 160, 166, or 167 amino acids in length.

[0123] The constriction region in the PorARc pore monomer or modified PorARc pore monomer s preferably from about 5 to about 114 amino acids in length. The constriction region in the modified PorARc pore monomer is preferably about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, or 114 amino acids in length. The constriction region in the PorARc pore monomer or modified PorARc pore monomer is preferably from about 15 to about 94 amino acids in length. The constriction region in the modified PorARc pore monomer is preferably about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, or 94 amino acids in length. The constriction region in the PorARc pore monomer or modified PorARc pore monomer is preferably about 15, 16, 17, 18, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 39, 50, 52, 86, 94 amino acids in length.

[0124] The PorARc pore monomer or modified PorARc pore monomer may comprise a transmembrane region. Before modification in accordance with the invention, the transmembrane region is preferably from about 15 to about 30 amino acids in length.

[0125] Before modification in accordance with the invention, the transmembrane region is preferably about 15, 16, 17 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 amino acids in length. The length of the transmembrane region may be increased in accordance with the invention as discussed in more detail below.

[0126] The PorARc pore is preferably selected from the pores in Table 1.

[0127] Table 1 - Different PorARc pores for use in the invention (aa = amino acids)

[0128] # Pore name Organism Reference Cap region Constricti Constricti (or scaffold) on region on [Length in [Length mutations aa] in aa]

[0129] 1. PorARc_Rco Rhodococcus WP_169818 1-86+107-180 87-106

[0130] corynebacteroide 371.1

[0160]

[0020] s

[0131]

[0132] PorARc_ Mycolicibacterium WP_061512 1-86+112-185 87-111 D91N / D92 Mph phlei 191.1

[0160]

[0025] N PorARc_Msp Mycobacterium MCB093076 1-86+113-127 87-112 D91N / D92 sp. 4.1

[0087]

[0026] N PorARc_Msp Mycobacterium MCB093526 1-86+121-194 87-120 D91N / D92 sp. 8.1

[0160]

[0034] N PorARc_Msp Mycobacterium WP_068199 1-86+115-188 87-114 D91N / D10 sp. 852014- 409.1

[0160]

[0028] 2N 52144

[0133] SCH5372336

[0134] PorARc_Msp Mycobacterium WP_071942 1-86+113-186 87-112 D91N / D92 sp. WY1O 913.1

[0160]

[0026] N PorARc_Msp Mycobacterium WP_067087 1-86+111-184 87-110 D91N sp. 852002- 132.1

[0160]

[0024]

[0135] 51961

[0136] SCH5331710

[0137] PorARc_Mrh Mycolicibacterium WP_014212 1-86+126-199 87-125 D91N / D92 rhodesiae 617.1

[0160]

[0039] N PorARc_Mph Mycolicibacterium WP_138248 1-86+118-191 87-117 D91N / D92 0 phocaicum 054.1

[0160]

[0031] N PorARc_Mm Mycolicibacterium WP_165605 1-86+117-190 87-116 D92N u mucogenicum 915.1

[0160]

[0030]

[0138] PorARc_Mch Mycolicibacterium WP_014817 1-86+116-189 87-115 D91N / D94 chubuense 420.1

[0160]

[0029] N / D104N PorARc_Mho Mycobacterium WP_069405 1-86+115-188 87-114 D91N holsaticum 343.1

[0160]

[0028]

[0139] PorARc_Mba Mycobacteriaceae MBY0291196 1-86+115-188 87-114 D91N bacterium.1

[0160]

[0028]

[0140] PorARc_Mch Mycolicibacterium WP_014814 1-86+114-187 87-113 D91N / D94 chubuense 623.1

[0160]

[0027] N / D102N PorARc_Mac Mycolicibacterium WP_064419 1-86+113-186 87-112 D91N / E10 acapulense 025.1

[0160]

[0026] 2Q PorARc_Mtu Mycolicibacterium WP_083126 1-86+110-183 87-109 D91N tusciae 605.1

[0160]

[0023]

[0141] PorARc_Mel Mycolicibacterium WP_128108 1-86+110-183 87-109 D91N / E10 elephantis 667.1

[0160]

[0023] IQ PorARc_Mfl Mycolicibacterium WP_069413 1-86+109-182 87-108 D92N flavescens 379.1

[0160]

[0022]

[0142] PorARc_Mco Mycolicibacterium WP_036397 1-86+108-181 87-107 D91N / D92 cosmeticum 228.1

[0160]

[0021] N PorARc_Ms Mycolicibacterium A0QR29 1-86+112-185 87-111 D91N / D92 m smegmatis

[0160]

[0027] N / D94N

[0143]

[0144] PorAR.c_R.sp Rhodococcus sp WP_065922 1-87+103-175 88-102 E90Q / D92 WB1 893.1

[0160]

[0015] N / D95N / D 100N PorARc_Rsp Rhodococcus sp WP_222638 1-86+102-175 87-101 D91N / D93

[0145] BP-316 310.1

[0160]

[0015] N / D99N PorARc_Rsp Rhodococcus sp WP_204866 1-86+102-175 87-101 D93N / D94

[0146] 430.1

[0160]

[0015] N / D99N PorARc_Rsp Rhodococcus sp WP_082833 1-86+102-175 87-101 D93N / D94

[0147] 904.1

[0160] N / D99N

[0015]

[0148] PorARc_Rsp unclassified WP_056447 1-86+103-176 87-102 E89Q / D91

[0149] Rhodococcus 532.1

[0160]

[0016] N / D93N / D 100N PorARc_Rsp Rhodococcus sp WP_222646 1-86+103-176 87-102 Q89E / D92

[0150] BP-241 231.1

[0160]

[0016] N / D94N PorARc_Rsp Rhodococcus sp WP_206003 1-87+106-178 88-105 D90N / D95

[0151] PSBB049 768.1

[0160]

[0018] N / D103N PorARc_Rsp Rhodococcus sp MBF0660864 1-86+104-176 87-103 Q89E / D91

[0152] .1

[0159]

[0017] N / D94N / D 101N PorARc_Rtr Rhodococcus WP_188546 1-86+103-176 87-102 E89Q / D92 trifolii 021.1

[0160]

[0016] N / D100N PorARc_Nm Nocardia WP_071926 1-86+103-175 87-102 E89Q / D91 a mangyaensis 418.1

[0159]

[0016] N / D93N / D 99N PorARc_Nsp Nocardia sp WP_225054 1-86+105-177 87-104 E89Q / E91 alder85J 379.1

[0159]

[0018] Q / D99N / D 102N PorARc_Aku Aldersonia WP_084435 1-86+105-177 87-104 D89N / E96 kunmingensis 981.1

[0159]

[0018] Q PorARc_Ncy Nocardia WP_138449 1-86+105-177 87-104 D89N / E91 cyriacigeorgica 037.1

[0159]

[0018] Q / D95N / D 102N PorARc_Nte Nocardia WP_082871 1-86+105-177 87-104 E89Q / D99 terpenica 519.1

[0159]

[0018] N / D102N PorARc_Gcr Gordonia crocea WP_161927 1-93 + 119-191 94-118

[0153] 960.1

[0166]

[0025]

[0154] PorARc_Rpy Rhodococcus WP_183079 1-86+173-245 87-172

[0155] pyridinivorans 242.1

[0159]

[0086]

[0156] PorARc_Rpy Rhodococcus UPW03596.1 1-86+181-253 87-180

[0157] pyridinivorans

[0159]

[0094]

[0158] PorARc_Aku Aldersonia WP_068276 1-86+105-177 87-104

[0159] kunmingensis 364.1

[0159]

[0018]

[0160]

[0161] PorARc_Nha Nocardia harenae WP_067652 1-86+107-180 87-106

[0162] 149.1

[0160]

[0020]

[0163] PorARc_Nfl Nocardia WP_063915 1-86+107-179 87-106

[0164] fla vo rosea 989.1

[0159]

[0020]

[0165] PorAR.c_R.sp Rhodococcus sp WP_094739 1-86+111-191 87-110

[0166] 14 14-2483-1-2 740.1

[0167]

[0024]

[0167] PorARc_Mfl Mycolicibacterium WP_187095 1-86+109-182 87-108 D91N fluoranthenivoran 255.1

[0160]

[0022]

[0168] s

[0169] PorARc_Mba Mycolicibacterium WP_197375 1-86+110-183 87-109

[0170] baixiangningiae 651.1

[0160]

[0023]

[0171] PorARc_Rru Rhodococcus WP_003937 1-87+108-180 88-107

[0172] B ruber 792.1

[0160]

[0020]

[0173] PorARc_Mw Mycolicibacterium WP_085146 1-86+109-182 87-108

[0174] 0 wolinskyi 114.1

[0160]

[0022]

[0175] PorARc_Mm Mycolicibacterium WP_163741 1-86+108-181 87-107

[0176] a madagascariense 319.1

[0160]

[0021]

[0177] PorARc_Msp Mycolicibacterium WP_155899 1-86+107-180 87-106 D91N / E96 sp CBMA 226 440

[0160]

[0020] Q PorARc_MII Mycolicibacterium WP_071289 1-86+107-180 87-106 D91N / E96 llatzerense 589

[0160]

[0020] Q PorARc_Mba Mycobacteriaceae NUS43967 1-86+107-180 87-106 D104N bacterium

[0160]

[0020]

[0178] PorARc_Mgi Mycolicibacterium WP_115328 1-86+108-181 87-107 D97N gilvum 206

[0160]

[0021]

[0179] PorARc_Nst Nocardia stercoris WP_122188 1-86+108-181 87-107 E98Q / D10

[0180] 535

[0160]

[0021] 5N PorARc_Msp Mycobacterium sp WP_056558 1-86+108-181 87-107 D91N / D92

[0181] Root 135 571

[0160]

[0021] N PorARc_Rca Rhodococcus WP_149428 1-86+108-181 87-107 E89Q / D91 cavernicola 944

[0160]

[0021] N / D102N PorARc_Mse Mycolicibacterium WP_163801 1-86+109-182 87-108 D91N / D92 sediminis 161

[0160]

[0022] N / E96Q PorARc_Mgo Mycolicibacterium MBU882976 1-86+109-182 87-108 D100N / D1 goodii 1

[0160]

[0022] 06N PorARc_Msp Mycolicibacterium WP_242453 1-86+109-182 87-108 E92Q sp P9-64 488

[0160]

[0022]

[0182] PorARc_Mm Mycolicibacterium BBZ30186 1-86+109-182 87-108 D91N / D92 a madagascariense

[0160]

[0022] N PorARc_Mfl Mycolicibacterium WP_069415 1-86+109-182 87-108 D91N / D92 flavescens 430

[0160]

[0022] N / D97N

[0183]

[0184] PorARc_Mm Mycolicibacterium WP_110316 1-86+109-182 87-108 D91N / D92 P moriokaense 655

[0160]

[0022] N PorARc_Mab Mycobacteroides WP_074269 1-86+109-182 87-108 - abscessus 525

[0160]

[0022]

[0185] PorARc_Mba Mycobacteriaceae MBV8966152 1-86+110-183 87-109 D91N bacterium

[0160]

[0023]

[0186] PorARc_Mva Mycolicibacterium ANI41733 1-86+110-183 87-109 D91N / D96 vaccae 95051

[0160]

[0023] N / D97N / D 101N PorARc_Rho unclassified WP_094631 1-86+110-183 87-109 E91Q / D97

[0187] Rhodococcus 231

[0160]

[0023] N / D102N /

[0188] D107N PorARc_Mtu Mycolicibacterium WP_083126 1-86+110-183 87-109 D91N tusciae 605

[0160]

[0023]

[0189] PorARc_Rsp Rhodococcus sp WP_128646 1-86+111-184 87-110 E91Q / D93

[0190] BS-15 324

[0160]

[0024] N / E99Q / D 108N PorARc_Msp Mycobacterium sp OBK75368 1-86+111-184 87-110 D91N 1164985.4

[0160]

[0024]

[0191] PorARc_Mga Mycobacterium BBY93710 1-86+112-185 87-111 D91N / D94 gallinarum

[0160]

[0025] N PorARc_Mba Mycobacteriaceae MBY0288091 1-86+112-185 87-111 D91N / D94 bacterium

[0160]

[0025] N PorARc_Mm Mycolicibacterium WP_152273 1-86+112-185 87-111 D91N / D92 u mucogenicum 437

[0160]

[0025] N PorARc_Mth Mycolicibacterium WP_040546 1-86+112-185 87-111 D91N / E92 thermoresistibile 596

[0160]

[0025] Q / D94N / D 103N PorARc_Mm Mycolicibacterium WP_197378 1-86+112-185 87-111 D91N / D95 e mengxianglii 436

[0160]

[0025] N PorARc_Mba Mycolicibacterium WP_193047 1-86+113-186 87-112 D91N / E94 baixiangningiae 902

[0160]

[0026] Q / E102Q PorARc_Msp Mycobacterium sp WP_167105 1-86+113-186 87-112 D91N DL592 159

[0160]

[0026]

[0192] PorARc_Mhe Mycolicibacterium WP_163751 1-86+113-186 87-112 D91N / D92 helvum 656

[0160]

[0026] N PorARc_Msp Mycobacterium sp MCB093826 1-86+113-186 87-112 D91N / D92

[0193] 5

[0160]

[0026] N / D101N PorARc_Msp Mycobacterium sp MCB129099 1-86+113-186 87-112 D91N / D92

[0194] 5

[0160]

[0026] N / D101N

[0195]

[0196] PorARc_Msp Mycobacterium sp PND57255 1-86+113-186 87-112 D91N / D92 ENV421

[0160]

[0026] N PorARc_Msp Mycolicibacterium WP_135423 1-86+113-186 87-112 D91N sp CH28 372

[0160]

[0026]

[0197] PorARc_Mau Mycolicibacterium WP_234884 1-86+113-186 87-112 D91N aubagnense 126

[0160]

[0026]

[0198] PorARc_Hsp Haiiangium sp WP_233206 1-86+113-186 87-112 D91N / D99

[0199] UPWRP 2 370

[0160]

[0026] N / D101N PorARc_Msp Mycobacterium sp WP_192727 1-86+113-186 87-112 D91N / D92

[0200] 069

[0160]

[0026] N PorARc_Mdi Mycobacterium WP_073859 1-86+113-186 87-112 E95Q / E97 diernhoferi 799

[0160]

[0026] Q / D101N /

[0201] D104N / D1 05N PorARc_Man Mycolicibacterium WP_163807 1-86+113-186 87-112 D91N anyangense 269

[0160]

[0026]

[0202] PorARc_Mph Mycolicibacterium WP_226520 1-86+113-186 87-112 D91N phocaicum 728

[0160]

[0026]

[0203] PorARc_Mrh Mycolicibacterium WP_083117 1-86+113-186 87-112 D91N rhodesiae 515

[0160]

[0026]

[0204] PorARc_Aba Actinomycetia MCH971032 1-86+114-187 87-113 D91N / E94 bacterium 7

[0160]

[0027] Q / E103Q PorARc_Mba Mycobacteriaceae OBA86335 1-86+114-187 87-113 D91N / D92 bacterium

[0160]

[0027] N / D100N 1482268.1

[0205] PorARc_Myc unclassified WP_239001 1-86+114-187 87-113 E103Q Mycobacteroides 602

[0160]

[0027]

[0206] PorARc_Msa Mycobacteroides WP_235629 1-86+114-187 87-113 E103Q / D1 saopaulense 490

[0160]

[0027] 04N PorARc_Msp Mycobacterium sp OBF25485 1-86+114-187 87-113 D91N / D94

[0207] ACS4331

[0160]

[0027] N / D102N PorARc_Mm Mycolicibacterium WP_234791 1-86+114-187 87-113 D91N u mucogenicum 467

[0160]

[0027]

[0208] PorARc_Msp Mycolicibacterium WP_155930 1-86+114-187 87-113 D91N / D92 sp CBMA 234 343

[0160]

[0027] N PorARc_Mps Mycolicibacterium WP_163721 1-86+114-187 87-113 D91N / D94 psychrotolerans 949

[0160]

[0027] N / D102N PorARc_Msp Mycobacterium sp WP_221370 1-86+114-187 87-113 D91N / D96

[0209] Y57 669

[0160]

[0027] N / D99N / D 105N PorARc_Msp Mycobacteroides WP_180751 1-86+115-188 87-114 E104Q sp LB1 945

[0160]

[0028]

[0210]

[0211] Mycobacterium sp WP_238885 1-86+115-188 87-114 D91N / D98 IDR2000157661 001

[0160]

[0028] N / E100Q /

[0212] D105N Mycobacterium WP_069405 1-86+115-188 87-114 D91N / holsaticum 343

[0160]

[0028] Mycolicibacterium WP_178360 1-86+115-188 87-114 D91N / E94 hippocampi 470

[0160]

[0028] Q / E98Q Mycobacterium sp WP_068199 1-86+115-188 87-114 D91N / D10 852014-52144 409

[0160]

[0028] 2N SCH5372336

[0213] unclassified WP_020111 1-86+115-188 87-114 D100N Rhodococcus 423

[0160]

[0028] Mycobacterium sp WP_101950 1-86+115-188 87-114 D91N / D92 3519 A 713

[0160]

[0028] N / D94N / D 100N / E101 Q / D112N Mycobacterium sp OBC05411 1-86+115-188 87-114 D91N / D92 852013-50091

[0160]

[0028] N SCH5140682

[0214] Mycolicibacterium WP_241402 1-86+115-188 87-114 D91N / D92 sp YH-1 605

[0160]

[0028] N Mycobacterium sp WP_068150 1-86+115-188 87-114 D91N E740 249

[0160]

[0028] Mycolicibacterium WP_128110 1-86+115-188 87-114 D91N / D10 elephantis 347

[0160]

[0028] IN Mycolicibacterium WP_138248 1-86+115-188 87-114 D91N / D92 phocaicum 053

[0160]

[0028] N Mycobacterium WP_220692 1-86+115-188 87-114 D91N / E98 holsaticum 273

[0160]

[0028] Q unclassified WP_222634 1-86+115-188 87-114 E105Q / D1 Rhodococcus 641

[0160]

[0028] 12N Mycolicibacterium WP_163721 1-86+115-188 87-114 D91N / D94 psychrotolerans 950

[0160]

[0028] N / D103N Mycolicibacterium WP_067395 1-86+115-188 87-114 D91N novocastrense 623

[0160]

[0028] Mycolicibacterium WP_163721 1-86+115-188 87-114 D91N / D94 psychrotolerans 948

[0160]

[0028] N / E99Q / D 103N Mycobacterium sp KUI25174 1-86+116-189 87-115 D91N / D94 IS-1742

[0160]

[0029] N / D101N /

[0215] E107Q

[0216]

[0217] PorARc_Mba Mycobacteriaceae MBV9090896 1-86+116-189 87-115 D91N bacterium

[0160]

[0029] PorARc_Msp Mycobacterium sp WP_221356 1-86+116-189 87-115 D91N / D94

[0218] PSTR-4-N 194

[0160]

[0029] N / D104N PorARc_Mha Mycolicibacterium WP_123766 1-86+116-189 87-115 D91N / D92 hassiacum 354

[0160]

[0029] N / D97N / D 98N / D107 N

[0219] PorARc_Msp Mycobacterium sp WP_064422 1-86+116-189 87-115 D91N / E10

[0220] GA-1285 115

[0160]

[0029] 2Q PorARc_Mau Mycolicibacterium WP_048634 1-86+116-189 87-115 D91N / D10 aurum 318

[0160]

[0029] 0N / D102N PorARc_Msp Mycobacterium sp TXH23533 1-86+116-189 87-115 D91N

[0160]

[0029]

[0221] PorARc_Msp Mycobacterium sp GFM17236 1-86+117-190 87-116 D91N / D99

[0222] PO1

[0160]

[0030] N / D100N /

[0223] D101N / D1 08N PorARc_Msp Mycolicibacterium GJF17981 1-86+117-190 87-116 D91N / E10 sp NGTWS0302

[0160]

[0030] 2Q / D105N / E106Q PorARc_Mau Mycolicibacterium WP_126316 1-86+117-190 87-116 D91N / D96 aurum 652

[0160]

[0030] N / D101N /

[0224] D103N PorARc_Msp Mycobacterium sp WP_135122 1-86+117-190 87-116 D91N / E93

[0225] PS03 16 788

[0160]

[0030] Q / E99Q / E1

[0226] 08Q PorARc_Msp Mycolicibacterium MBX7450768 1-86+117-190 87-116 D91N / D94 sp 3033

[0160]

[0030] N / D99N / D 105N PorARc_Msp Mycobacterium sp MCB092883 1-86+117-190 87-116 D91N / D10

[0227] 5

[0160]

[0030] 5N PorARc_Msp Mycobacterium sp WP_066833 1-86+117-190 87-116 D91N / E94

[0228] 852013-51886 589

[0160]

[0030] Q / D100N / SCH5428379 E102Q / E10

[0229] 6Q PorARc_Mli Mycolicibacterium BBY16945 1-86+117-190 87-116 D91N / E93 litorale

[0160]

[0030] Q / D96N / E 108Q PorARc_Mm Mycolicibacterium WP_061010 1-86+117-190 87-116 D91N / D92 u mucogenicum 468

[0160]

[0030] N

[0230]

[0231] PorARc_Mm Mycolicibacterium WP_234791 1-86+117-190 87-116 D92N u mucogenicum 471

[0160]

[0030]

[0232] PorARc_Mw Mycolicibacterium WP_085146 1-86+117-190 87-116 D91N / D92 0 wolinskyi 113

[0160]

[0030] N PorARc_Msp Mycolicibacterium GJF17985 1-86+117-190 87-116 D91N / E10 sp NGTWS0302

[0160]

[0030] 2Q / D105N / E106Q PorARc_Mm Mycolicibacterium OBB58307 1-86+117-190 87-116 D91N / D94 0 monacense

[0160]

[0030] N / D99N / D 101N / E108 Q

[0233] PorARc_Mm Mycolicibacterium WP_064916 1-86+117-190 87-116 D91N / E93 0 monacense 949

[0160]

[0030] Q / D96N PorARc_Msp Mycobacterium sp MCF6390467 1-86+118-191 87-117 D91N / D94

[0234] MBM

[0160]

[0031] N / D104N /

[0235] D106N PorARc_Aba Actinobacteria MBN963680 1-86+118-191 87-117 D91N / E96 bacterium 1

[0160]

[0031] Q PorARc_Msp Mycolicibacterium WP_207361 1-86+118-191 87-117 D91N / D94 sp S2-37 462

[0160]

[0031] N / D100N /

[0236] D101N / D1 04N / E109 Q

[0237] PorARc_Mne Mycolicibacterium WP_019510 1-86+118-191 87-117 D93N / E11 neoaurum 421

[0160]

[0031] 0Q PorARc_Msp Mycobacterium sp WP_135122 1-86+118-191 87-117 D91N / E94

[0238] PS03 16 786

[0160]

[0031] Q / E100Q /

[0239] D101N / D1 04N / E109 Q

[0240] PorARc_Mpo Mycolicibacterium WP_163678 1-86+118-191 87-117 D91N / E98 poriferae 668

[0160]

[0031] Q / E100Q /

[0241] D102N / D1 09N PorARc_Mpa Mycobacterium WP_099025 1-86+118-191 87-117 D91N / D94 palauense 911

[0160]

[0031] N / D102N /

[0242] D106N PorARc_Msp Mycolicibacterium WP_217889 1-86+118-191 87-117 D91N / D96 sp PAM1 399

[0160]

[0031] N / D101N /

[0243] D102N / D1 09N PorARc_Myc Mycolicibacterium WP_115280 1-86+119-192 87-118 D91N / D94

[0244] 310

[0160]

[0032] N / D107N

[0245]

[0246] PorARc_Mbr Mycolicibacterium WP_128111 1-86+119-192 87-118 D91N / D98 brumae 797

[0160]

[0032] N / E99Q / D 100N PorARc_Msp Mycobacterium sp WP_135122 1-86+119-192 87-118 D91N / E94

[0247] PS03 16 787

[0160]

[0032] Q / E99Q / D 100N / D10 1N / E110Q PorARc_Mfa Mycolicibacterium WP_085099 1-86+119-192 87-118 D91N / E10 fallax 073

[0160]

[0032] 2Q / D106N / E110Q PorARc_Msp Mycolicibacterium WP_242638 1-86+119-192 87-118 D91N / D94 sp S2-37 988

[0160]

[0032] N / D100N /

[0248] D101N / D1 04N / E110 Q

[0249] PorARc_Mfr Mycolicibacterium MBX9919851 1-86+120-193 87-119 D91N / D94 frederiksbergense

[0160]

[0033] N / D99N / D 108N PorARc_Rsp Rhodococcus sp WP_094719 1-86+120-193 87-119 D95N / D10

[0250] 14-2470-lb 388

[0160]

[0033] 1N / E104Q /

[0251] D110N PorARc_Mto Mycolicibacterium STZ61066 1-86+120-193 87-119 D91N / D94 tokaiense

[0160]

[0033] N / D108N PorARc_Msp Mycobacterium sp WP_083741 1-86+120-193 87-119 D91N / D94

[0252] MS 1601 162

[0160]

[0033] N / D99N / D 104N / D10 8N PorARc_Msp Mycobacterium sp ANW65896 1-86+120-193 87-119 D91N / D94 dJI-10

[0160]

[0033] N / D104N /

[0253] D108N PorARc_Msp Mycobacterium sp WP_059093 1-86+120-193 87-119 D91N / D94

[0254] GA-2829 044

[0160]

[0033] N / D105N /

[0255] D108N / E1 12Q PorARc_Mm Mycolicibacterium WP_197380 1-86+121-194 87-120 D91N / D94 e mengxianglii 164

[0160]

[0034] N / D99N / D 106N / D10 9N / E113Q PorARc_Mdo Mycobacterium WP_085188 1-86+122-195 87-121 D91N / E94 doricum 080

[0160]

[0035] Q / E99Q / D 104N / D10 8N / E113Q

[0256]

[0257] 154. PorARc_Mab Mycobacteroides WP_234804 1-86+122-195 87-121 E111Q abscessus 504

[0035] 155. PorARc_Mrh Mycolicibacterium WP_014212 1-86+126-199 87-125 D91N / D92 rhodesiae 617

[0160]

[0039] N

[0258] 156. PorARc_Mm Mycolicibacterium WP_083149 1-86+126-199 87-125 D91N / D92 0 moriokaense 007

[0160]

[0039] N / D98N 157. PorARc_Mch Mycobacteroides MBB4856568 1-86+137-210 87-136 E126Q chelonae

[0160]

[0050] 158. PorARc_Mab Mycobacteroides WP_032692 1-86+139-212 87-138 E107Q / E12 abscessus 145

[0160]

[0052] 8Q 159. PorARc_Msa Mycobacteroides TDZ98543 1-86+139-212 87-138 E107Q / D1 salmoniphilum

[0160]

[0052] 24N / E128

[0259] Q

[0260] 160. PorARc_Myc unclassified WP_155940 1-86+139-212 87-138 E107Q / E12

[0261] Mycobacteroides 737

[0160]

[0052] 8Q 161. PorARc_Mch Mycobacteroides OHU80649 1-86+139-212 87-138 E107Q / E12 chelonae

[0160]

[0052] 8Q / D129N

[0262]

[0263] The PorARc pore is preferably selected from the pores in Table 1. The (fourth) Reference column provides a reference to the wild-type (or naturally occurring) sequence of each pore on GenBank. The PorARc pore may be selected from any of the wild-type pores in Table 1 ( / .e., from the references in the fourth column). The PorARc pore may be selected from any of the wild-type pores in Table 1 with the signal peptide removed and a methionine (M) at the N terminus ( / .e., at position 1). The skilled person can determine these sequences from the references in the fourth column.

[0264] Preferred cap and constriction regions for each pore are shown in the fifth and sixth columns of Table 1. The residue numbering in the fifth and sixth columns correspond to the wild-type sequences without the signal peptide and having a methionine (M) at the N terminus ( / .e., at position 1). The residue numbering of the cap and constriction regions will need to be adjusted for the wild-type (or naturally occurring) sequences in the references in the fourth column. The cap and constriction regions shown in the fifth and sixth columns of Table 1 are preferred cap and constriction regions. The invention covers additional cap and constriction regions which differ from those shown in the fifth and sixth columns by ± about 10 amino acids. For instance, in PorARc_Rco (pore 1):

[0265] 1-86 includes 1-76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95 or 96;107-180 includes 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116 or 117-170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189 or 190

[0266] 87-106 includes 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97-96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116 or 117.

[0267] This equally applies to pores 2-161 in Table 1. As explained in more detail below, the modified pore monomer preferably comprises one or more modifications which stabilise a pore formed from the modified pore monomer and / or improve the ability of a pore formed from the modified pore monomer to characterise a target analyte. One or more negatively charged amino acids in any of the pores in Table 1 may be removed, for instance by deletion or substitution. One or more negatively charged amino acids in any of the pores in Table 1, such as one or more E and / or D, are preferably deleted or substituted with one or more different amino acids, such as one or more positively charged amino acids and / or one or more uncharged amino acids. This removes negative charge from the sequence of the pore. Any number of negatively charged amino acids may be deleted or substituted, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. The one or more negatively charged amino acids are preferably in the constriction region of the pore. The preferred constriction region for each pore is defined in sixth column of Table 1. This applies to any of the sequences in Table 1, including the wild-type sequences and the wild-type sequences without the signal peptide and having a methionine (M) at the N terminus ( / .e., at position 1).

[0268] Preferred constriction substitutions for each pore are shown in the seventh column. The PorARc pore may be selected from any of the pores in Table 1 comprising substitution of the negatively charged amino acid, such as D or E, with a positively charged amino acid, such as R, H or K, or an uncharged amino acid, such as S, T, N or Q, at one or more of the positions shown, preferably all of the positions shown. For instance, the PorARc pore may be pore 2 with substitution of D with a positively charged amino acid, such as R, H or K, or an uncharged amino acid, such as S, T, N or Q, at position 91 and / or position 92. The PorARc pore may be pore 3, pore 8 or pore 19 with substitution of D with a positively charged amino acid, such as R, H or K, or an uncharged amino acid, such as S, T, N or Q, at position 91 and / or position 92. The PorARc pore may be pore 17 with substitution of D or E with a positively charged amino acid, such as R, H or K, or an uncharged amino acid, such as S, T, N or Q, at position 91 and / or position 101. The PorARc pore may be pore 25 with substitution of D or E with a positively charged amino acid, such as R, H or K, or an uncharged amino acid, such as S, T, N or Q, at one or more of, preferably all of, position 89, 91, 93 and 100. The PorARc pore may be pore 27 with substitution of D with a positively charged amino acid, such as R, H or K, or an uncharged amino acid, such as S, T, N or Q, atone or more of, preferably all of, positions 90, 95 and 103. This applies to any of the sequences in Table 1, including the wild-type sequences and the wild-type sequences without the signal peptide and having a methionine (M) at the N terminus ( / .e., at position 1). The residue numbering in the seventh column corresponds to the wild-type sequences without the signal peptide and having a methionine (M) at the N terminus ( / .e., at position 1).

[0269] The PorARc pore may be selected from any of the pores in Table 1 comprising one or more of, or all of, the substitutions shown in the seventh column. For instance, the PorARc pore may be pore 2 in Table 1 with D91N and / or D92N. The PorARc pore may be pore 3, pore 8 or pore 19 with D91N and / or D92N. The PorARc pore may be pore 17 with D91N and / or E101Q. The PorARc pore may be pore 25 with one or more of, preferably all of, E89Q, D91N, D93N and D100N. The PorARc pore may be pore 27 with one or more of, preferably all of, D90N, D95N and D103N. This applies to any of the sequences in Table 1, including the wild-type sequences and the wild-type sequences without the signal peptide and having a methionine (M) at the N terminus ( / .e., at position 1). The residue numbering in the seventh column corresponds to the wild-type sequences without the signal peptide and having a methionine (M) at the N terminus ( / .e., at position 1).

[0270] The PorARc pore may be pore 1 in Table 1 with one or more cap substitutions. For instance, the PorARc is preferably pore 1 in Table 1 with substitution of D or E with a positively charged amino acid, such as R, H or K, or an uncharged amino acid, such as S, T, N or Q, at one or more of, preferably all of, positions 78, 82, 116, 125 and 165. The PorARc pore is preferably pore 1 in Table 1 with one or more of, preferably all of, E78R, D82S, E116T, E125A, or D165S. This applies to any of the pore 1 sequences in Table 1, including the wildtype sequence and the wild-type sequences without the signal peptide and having a methionine (M) at the N terminus ( / .e., at position 1). The PorARc pore preferably comprises or consists of the sequence shown in SEQ ID NO: 1.

[0271] The PorARc pore may be pore 1 in Table 1 with one or more constriction substitutions. For instance, the PorARc is preferably pore 1 in Table 1 with substitution of D or E with a positive amino acid, such as R, H or K, or an uncharged amino acid, such as S, T, N or Q, at position 89 and / or position 104. The PorARc pore is preferably pore 1 in Table 1 with E89R, E89Q, E89L or E89A and / or D104S or D104N, such as E89R / D104S, E89Q / D104S, E89Q / D104N, E89L / D104S, E89L / D104N, E89A / D104S or E89A / D104N. The PorARc pore is preferably pore 1 in Table 1 with E89R and / or D104S, such as E89R / D104S. This applies to any of the pore 1 sequences in Table 1, including the wild-type sequence and the wild-type sequences without the signal peptide and having a methionine (M) at the N terminus ( / .e., at position 1). These one or more constriction substitutions may be made in addition to the one or more cap substitutions discussed above.Preferred PorARc pore monomers for use in the invention are shown in SEQ ID NOs: 1-8. The PorARc pore monomer and / or the modified PorARc pore monomer is preferably a structural variant having a root mean square deviation (RMSD) of less than about 4.0 Angstroms (Å), such as less than about 3.5 Å, less than about 3.0 Å, less than about 2.5 Å, less than about 2.0 Å, less than about 1.5 Å, less than about 1.0 Å or less than about 0.5 Å, when compared with any of the sequences shown in SEQ ID NOs: 1-8.

[0272] The PorARc pore monomer and / or the modified PorARc pore monomer preferably comprises a variant sequence having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity to any of the sequences shown in SEQ ID NOs: 1-8. The PorARc pore monomer and / or the modified PorARc pore monomer preferably comprises a variant sequence having at least about 80% homology and / or identity to any of the sequences shown in SEQ ID NOs: 1-8. The PorARc pore monomer and / or the modified PorARc pore monomer preferably comprises a sequence having at least about 90% homology and / or identity to any of the sequences shown in SEQ ID NOs: 1-8. The PorARc pore monomer preferably comprises a sequence having 100% homology and / or identity to any of the sequences shown in SEQ ID NOs: 1-8. Homology and / or identity is typically measured over the entire length of the monomer. Variants may have modifications at any of the positions and / or any of the substitutions disclosed in WO 2023 / 118404 and WO 2024 / 089270 (incorporated by reference herein in their entireties).

[0273] Sequence homology and / or identity can also relate to a fragment or portion of the reference sequence. Hence, a sequence may have less than 40% overall sequence homology and / or identity with any of the sequences shown in SEQ ID NOs: 1-8, but the sequence of a particular region, domain or subunit could share at least about 80%, at least about 90%, or as much as about 99% sequence homology and / or identity with the corresponding region of the wild-type or native monomer. There may be at least about 80%, at least 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity over a stretch of 100 or more, for example 125, 150, 175 or 200 or more, contiguous amino acids ("hard homology").

[0274] A PorARr pore monomer is a monomer that is capable of forming a PorARr pore. Such monomers are known in the art, especially from WO 2023 / 118404 (incorporated by reference herein in its entirety). The PorARr pore monomer or modified PorARr pore monomer preferably comprises a cap region (or scaffold) and a constriction region. The PorARr pore monomer or modified PorARr pore monomer preferably comprises one or more of (a) a cap region, (b) a constriction region, and (c) a transmembrane beta barrel region,such as (a), (b), (c), (a) and (b), (a) and (c), (b) and (c), or (a), (b) and (c). The cap region and the transmembrane region may together be known as a scaffold. The PorARr pore monomer or modified PorARr pore monomer preferably comprises (a)-(c).

[0275] A preferred PorARr pore monomer for use in the invention is shown in SEQ ID NO: 9. The PorARr pore monomer and / or the modified PorARr pore monomer is preferably a structural variant having a root mean square deviation (RMSD) of less than about 4.0 Angstroms (Å), such as less than about 3.5 Å, less than about 3.0 Å, less than about 2.5 Å, less than about 2.0 Å, less than about 1.5 Å, less than about 1.0 Å or less than about 0.5 Å, when compared with the sequence shown in SEQ ID NO: 9.

[0276] The PorARr pore monomer and / or the modified PorARr pore monomer preferably comprises a variant sequence having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity to the sequence shown in SEQ ID NO: 9. The PorARr pore monomer and / or the modified PorARr pore monomer preferably comprises a variant sequence having at least about 80% homology and / or identity to the sequence shown in SEQ ID NO: 9. The PorARr pore monomer and / or the modified PorARr pore monomer preferably comprises a sequence having at least about 90% homology and / or identity to the sequence shown in SEQ ID NO: 9. The PorARr pore monomer preferably comprises a sequence having 100% homology and / or identity to the sequence shown in SEQ ID NO: 9. Homology and / or identity is typically measured over the entire length of the monomer. Variants may have modifications at any of the positions and / or any of the substitutions disclosed in WO 2023 / 118404 and WO 2024 / 089270 (incorporated by reference herein in their entireties).

[0277] Sequence homology and / or identity can also relate to a fragment or portion of the reference sequence. Hence, a sequence may have less than 40% overall sequence homology and / or identity with the sequence shown in SEQ ID NO: 9, but the sequence of a particular region, domain or subunit could share at least about 80%, at least about 90%, or as much as about 99% sequence homology and / or identity with the corresponding region of the wild-type or native monomer. There may be at least about 80%, at least 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity over a stretch of 100 or more, for example 125, 150, 175 or 200 or more, contiguous amino acids ("hard homology").

[0278] A PorBRr pore monomer is a monomer that is capable of forming a PorBRr pore. Such monomers are known in the art, especially from WO 2023 / 118404 (incorporated by reference herein in its entirety). The PorBRr pore monomer or modified PorBRr poremonomer preferably comprises a cap region (or scaffold) and a constriction region. The PorBRr pore monomer or modified PorBRr pore monomer preferably comprises one or more of (a) a cap region, (b) a constriction region, and (c) a transmembrane beta barrel region, such as (a), (b), (c), (a) and (b), (a) and (c), (b) and (c), or (a), (b) and (c). The cap region and the transmembrane region may together be known as a scaffold. The PorBRr pore monomer or modified PorBRr pore monomer preferably comprises (a)-(c).

[0279] A preferred PorBRr pore monomer for use in the invention is shown in SEQ ID NO: 10. The PorBRr pore monomer and / or the modified PorBRr pore monomer is preferably a structural variant having a root mean square deviation (RMSD) of less than about 4.0 Angstroms (Å), such as less than about 3.5 Å, less than about 3.0 Å, less than about 2.5 Å, less than about 2.0 Å, less than about 1.5 Å, less than about 1.0 Å or less than about 0.5 Å, when compared with the sequence shown in SEQ ID NO: 10.

[0280] The PorBRr pore monomer and / or the modified PorBRr pore monomer preferably comprises a variant sequence having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity to the sequence shown in SEQ ID NO: 10. The PorBRr pore monomer and / or the modified PorBRr pore monomer preferably comprises a variant sequence having at least about 80% homology and / or identity to the sequence shown in SEQ ID NO: 10. The PorBRr pore monomer and / or the modified PorBRr pore monomer preferably comprises a sequence having at least about 90% homology and / or identity to the sequence shown in SEQ ID NO: 10. The PorBRr pore monomer preferably comprises a sequence having 100% homology and / or identity to the sequence shown in SEQ ID NO: 10. Homology and / or identity is typically measured over the entire length of the monomer. Variants may have modifications at any of the positions and / or any of the substitutions disclosed in WO 2023 / 118404 and WO 2024 / 089270 (incorporated by reference herein in their entireties).

[0281] Sequence homology and / or identity can also relate to a fragment or portion of the reference sequence. Hence, a sequence may have less than 40% overall sequence homology and / or identity with the sequence shown in SEQ ID NO: 10, but the sequence of a particular region, domain or subunit could share at least about 80%, at least about 90%, or as much as about 99% sequence homology and / or identity with the corresponding region of the wild-type or native monomer. There may be at least about 80%, at least 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity over a stretch of 100 or more, for example 125, 150, 175 or 200 or more, contiguous amino acids ("hard homology").MspA, MspB and MspC

[0282] The pore monomer is preferably from or derived from MspA, MspB or MspC. The modified pore monomer is preferably from or derived from MspA, MspB or MspC. The actinobacterial pore monomer or the modified actinobacterial pore monomer is preferably from or derived from MspA, MspB or MspC. The pore monomer is preferably from or derived from MspA. The modified pore monomer is preferably from or derived from MspA. The actinobacterial pore monomer or the modified actinobacterial pore monomer is preferably from or derived from MspA. The pore monomer, the modified pore monomer, the actinobacterial pore monomer or the modified actinobacterial pore monomer may be from or derived from MspB or MspC.

[0283] The terms "from" and "derived from" are defined above. The pore monomer or the modified pore monomer may be, may be from or may be derived from any of the MspA pores described in WO 2012 / 107778, WO 2015 / 166275, WO 2016 / 055778, WO 2016 / 132124, and WO 2016 / 132123 (all incorporated herein by reference in their entireties) or a variant thereof.

[0284] In discussion below, pore monomers from or derived from MspA, MspB or MspC may be referred to as MspA, MspB or MspC pore monomers respectively. They may be MspA, MspB or MspC pore monomers before modification in accordance with the invention, e.g., MspA pore monomer. They may be MspA, MspB or MspC pore monomers modified in accordance with the invention, e.g., "modified" MspA pore monomer.

[0285] In all of the discussion below relating to increasing a distance, the terms "pore monomer" and "modified pore monomer" preferably relate to "a MspA, MspB or MspC pore monomer" or "a modified MspA, MspB or MspC pore monomer" respectively.

[0286] The MspA, MspB or MspC pore preferably comprises a cap region (or scaffold) and a constriction region. The MspA, MspB or MspC pore preferably comprises one or more of (a) a cap region, (b) a constriction region, and (c) a transmembrane beta barrel region, such as (a), (b), (c), (a) and (b), (a) and (c), (b) and (c), or (a), (b) and (c). The cap region and the transmembrane region may together be known as a scaffold. The MspA, MspB or MspC pore preferably comprises (a)-(c). These regions in MspA are shown in Figure 2.

[0287] A MspA monomer is capable of forming a MspA pore. Before modification in accordance with the invention, the MspA pore may be any size but preferably has the dimensions of the wildtype MspA (Figure 2). The MspA pore preferably has an external diameter of from about 70 to about 110 A at its widest point, such as from about 80 to about 100 A or from about 85 to about 95 A at its widest point. The MspA pore preferably has an external diameter of about 92 A at its widest point. The MspA pore preferably has a total length of from about 70 to about 110 A, such as from about 80 to about 100 A or from about 85 to about 95 A. TheMspA pore preferably has a total length of about 90 A. References to "total length" and "length" relate to the length of the pore or pore region when viewed from the side (see, e.g., the side view in Figure 2).

[0288] The cap region (A in Figure 2) preferably has a length of from about 25 to about 65 A, such as from about 35 to about 55 A or from about 40 to about 50 A. The cap region preferably has a length of about 48 A. The channel defined by the cap region preferably has an opening of from about 30 to about 70 A in diameter, such as from about 40 to about 60 A or from about 45 to about 55 A in diameter. The channel defined by the cap region preferably has an opening of about 53 A in diameter. The channel defined by the cap region is preferably from about 20 to about 60 A in diameter at its narrowest point, such as from about 30 to about 55 A or from about 35 to about 50 A in diameter at its narrowest point. The channel defined by the cap region is preferably about 47 A in diameter at its narrowest point.

[0289] The transmembrane beta barrel region (B in Figure 2) preferably has a length of from about 5 to about 45 A, such as from about 15 to about 35 A or from about 20 to about 30 A. The transmembrane beta barrel preferably has a length of about 24 A. The channel defined by the transmembrane beta barrel region is preferably from about 20 to about 60 A in diameter at its narrowest point, such as from about 30 to about 50 A or from about 35 to about 45 A in diameter at its narrowest point. The channel defined by the transmembrane beta barrel region is preferably about 41 A in diameter at its narrowest point. The transmembrane region is preferably from about 30 to about 80 A in diameter, such as from about 40 to about 75 A or from about 45 to about 65 A. The constriction region is preferably about 63 A in diameter.

[0290] The cap region (or scaffold) (C in Figure 2 and formed from A and B) preferably has a length of from about 55 to about 95 A, such as from about 65 to about 85 A or from about 70 to about 80 A. The cap region (or scaffold) preferably has a length of about 72 A. The channel defined by the cap region (or scaffold) (C) is preferably from about 20 to about 60 A in diameter at its narrowest point, such as from about 30 to about 50 A or from about 35 to about 45 A in diameter at its narrowest point. The channel defined by the cap region (or scaffold) (C) is preferably about 41 A in diameter at its narrowest point.

[0291] The constriction region (D in Figure 2) preferably has a length of from about 5 to about 40 A, such as from about 10 to about 30 A or from about 15 to about 25 A. The constriction region preferably has a length of about 19 A. The channel defined by the constriction region is preferably from about 10 to about 50 A in diameter at its narrowest point, such as from about 12 to about 40 A, from about 15 to about 32 A or from about 18 to about 35 A in diameter at its narrowest point. The channel defined by the constriction region is preferablyabout 20 A in diameter at its narrowest point. The channel defined by the constriction region is preferably from about 10 to about 60 A in diameter at its narrowest point, such as from about 15 to about 55 A, from about 25 to about 50 A or from about 30 to about 45 A in diameter at the base of the pore structure. The channel defined by the constriction region is preferably about 43 A in diameter at the base of the pore structure.

[0292] All of the measurements above are based on measuring from backbone to backbone of the amino acids forming the different regions (as shown in Figure 2). All of the measurements typically apply to the PorARc pore monomer or PorARc pore before it is modified in accordance with the invention.

[0293] A MspA pore monomer is a monomer that is capable of forming a MspA pore. Such monomers are known in the art, especially from WO 2023 / 118404 (incorporated by reference herein in its entirety). The MspA pore monomer or modified MspA pore monomer preferably comprises a cap region (or scaffold) and a constriction region. The MspA pore monomer or modified MspA pore monomer preferably comprises one or more of (a) a cap region, (b) a constriction region, and (c) a transmembrane beta barrel region, such as (a), (b), (c), (a) and (b), (a) and (c), (b) and (c), or (a), (b) and (c). The cap region and the transmembrane region may together be known as a scaffold. The MspA pore monomer or modified MspA pore monomer preferably comprises (a)-(c). These regions in MspA are shown in Figure 2.

[0294] Preferred MspA pore monomers for use in the invention are shown in SEQ ID NO: 11 or 12. The MspA pore monomer and / or the modified MspA pore monomer is preferably a structural variant having a root mean square deviation (RMSD) of less than about 4.0 Angstroms (A), such as less than about 3.5 A, less than about 3.0 A, less than about 2.5 A, less than about 2.0 A, less than about 1.5 A, less than about 1.0 A or less than about 0.5 A, when compared with the sequence shown in SEQ ID NO: 11 or 12.

[0295] The MspA pore monomer and / or the modified MspA pore monomer preferably comprises a variant sequence having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity to the sequence shown in SEQ ID NO: 11 or 12. The MspA pore monomer and / or the modified MspA pore monomer preferably comprises a variant sequence having at least about 80% homology and / or identity to the sequence shown in SEQ ID NO: 11 or 12. The MspA pore monomer and / or the modified MspA pore monomer preferably comprises a sequence having at least about 90% homology and / or identity to the sequence shown in SEQ ID NO: 11 or 12. The MspA pore monomer preferably comprises a sequence having100% homology and / or identity to the sequence shown in SEQ ID NO: 11 or 12. Homology and / or identity is typically measured over the entire length of the monomer. Variants may have modifications at any of the positions and / or any of the substitutions disclosed in WO 2023 / 118404 and WO 2024 / 089270 (incorporated by reference herein in their entireties).

[0296] Sequence homology and / or identity can also relate to a fragment or portion of the reference sequence. Hence, a sequence may have less than 40% overall sequence homology and / or identity with the sequence shown in SEQ ID NO: 11 or 12, but the sequence of a particular region, domain or subunit could share at least about 80%, at least about 90%, or as much as about 99% sequence homology and / or identity with the corresponding region of the wildtype or native monomer. There may be at least about 80%, at least 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity over a stretch of 100 or more, for example 125, 150, 175 or 200 or more, contiguous amino acids ("hard homology").

[0297] In any of these structural or sequence variants of SEQ ID NO: 12, the amino acid at the position corresponding to position 93 in SEQ ID NO: 12 can be aspartic acid (D).

[0298] The MspA pore monomer or modified MspA pore monomer may comprise one or more mutations, such as one or more additions, one or more substitutions and / or one or more deletions. The MspA pore monomer or modified MspA pore monomer, including any of the structural variants or variant sequences discussed above, preferably comprises the following substitutions: D90N, D91N, D93N, D118R, D134R and E139K wherein the position numbering corresponds to SEQ ID NO: 11. The monomer may comprise a structural variant and / or variant sequence of SEQ ID NO: 11 comprising D90N, D91N, D93N, D118R, D134R and E139K. The variant sequence may have any of the percentages of homology and / or identity set out above. Preferred substitutions are disclosed in WO 2012 / 107778 (hereby incorporated by reference in its entirety). Particularly preferred variants include, but are not limited to, those including the following substitution(s) with the position numbering corresponding to SEQ ID NO: 11: L88N; L88S; L88Q; L88T; D90S; D90Q; D90Y; I105L; I105S; Q126R; G75S; G77S; G75S, G77S, L88N / Q126R; G75S, G77S, L88N, D90Q / Q126R; D90Q / Q126R; L88N, D90Q / Q126R; L88S / D90Q; L88N / D90Q; E59R; G75Q; G75N; G75S; G75T; G77Q; G77N; G77S; G77T; I78L; S81N; T83N; N86S; N86T; I87F; I87V; I87L;

[0299] L88N; L88S; L88Y; L88F; L88V; L88Q; L88T; I89F; I89V; I89L; N90S; N90Q; N90L; N90Y; N91S; N91Q; N91L; N91M; N91I; N91A; N91V; N91G; G92A; G92S; N93S; N93A; N93T; I94L; T95V; A96R; A96D; A96V; A96N; A96S; A96T; P97S; P98S; F99S; G100S; L101F; N102; N102S; N102T; S103A; S103Q; S103N; S103G; S103T; V104I; I105Y; I105L;

[0300] I105A; I105G; I105Q; I105N; I105S; I105T; T106F; T106I; T106V; T106S; N108P; N108S; D90Q / I105A; D90S / G92S; L88T / D90S; I87Q / D90S; I89Y / D90S; L88N / I89F; L88N / I89Y; D90S / G92A; D90S / I94N; D90S / V104I; L88D / I105K; L88N / Q126R; L88N, D90Q / D91R;L88N, D90Q / D91S; L88N, D90Q / I105V; D90Q, D93S / I105A; N91Y; N90Y / N91G; N90G / 91Y; N90G / N91G; 105G; N90R; N91R; N90R / N91R; N90K; N91; N90K / N91K; N90Q / 91G;

[0301] N90G / 91Q; N90Q / 91Q; R118N; N91C; N90C; N90W; N91W; N90K; N91K; N90R; N91R; N90S / N91S; N90Y / I105A; N90G / I105A; N90Q / I105A; N90S / I105A; L88A / I105A;

[0302] L88S / I105S; L88N / I105N; N90G / 93G; N90G; N93G; N90G / 91A; I105K; I105R; I105V; I105P; I105W; L88R; L88A; L88G; L88N; N90R / I105A; N90S / I105A; L88A / I105A;

[0303] L88S / I105S; L88N / I105N; L88C; S103C; I105C; and D134R.

[0304] Preferred chimeric pore monomers

[0305] The pore monomer or the modified pore monomer may be from or derived from a chimeric pore monomer comprising two or more regions, wherein at least two of the two or more regions are from at least two different actinobacterial pores. The at least two different actinobacterial pores may be selected from PorARc, PorARr, PorBRr, MspA, MspB, MspC and MspD. The chimeric pore monomer is capable of forming a pore. This can be measured using routine methods including any of those discussed above. The chimeric pore monomer may be any of those described in WO 2024 / 089270 (incorporated by reference herein in its entirety).

[0306] The chimeric pore monomer comprises two or more regions. The chimeric pore monomer may comprise any number of regions, such as three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more or ten or more regions. The chimeric pore monomer may comprise three, four, five, six, seven, eight, nine or ten regions. The chimeric pore monomer preferably comprises two regions. The chimeric pore monomer preferably comprises three regions. The chimeric pore monomer preferably comprises five regions.

[0307] Specific regions, such as the cap region or constriction region, in a pore may be identified using standard methods. This is discussed above.

[0308] The at least two regions preferably comprise a cap region and a constriction region.

[0309] The cap region (or scaffold) in the chimeric pore monomer may be longer, shorter or the same length as the cap region (or scaffold) in the pore which the constriction region in the chimeric pore monomer is from or derived from. The constriction region in the chimeric pore monomer may be longer, shorter or the same length as the constriction region in the pore which the cap region (or scaffold) in the chimeric pore monomer is from or derived from. The constriction region in the chimeric pore monomer is preferably shorter than the constriction region in the pore which the cap region (or scaffold) in the chimeric pore monomer is from or derived from. Length may be measured in terms of amino acid number and / or length along the sagittal (or longitudinal) plane of the pore.The at least two regions preferably comprise a cap region, a constriction region, and a transmembrane region. The transmembrane region may be a transmembrane beta barrel region or a transmembrane alpha helical region. The transmembrane region is preferably a transmembrane beta barrel region. CsgG pores typically comprise these three regions and they are discussed in more detail below in relation to CsgG pores. In these embodiments with a cap region, a constriction region, and a transmembrane region, the cap region and transmembrane region together is also known as a scaffold.

[0310] The cap region may further comprise two subregions, namely a landing platform region and a carboxy-terminal (C-terminal) region. The at least two regions preferably comprise a cap region, a landing platform region, a C-terminal region, a constriction region, and a transmembrane region. The transmembrane region may be a transmembrane beta barrel region or a transmembrane alpha helical region. The transmembrane region is preferably a transmembrane beta barrel region. CsgG pores typically comprise these five regions and they are discussed in more detail below in relation to CsgG pores. In these embodiments, the cap region, the landing platform region, the C-terminal region and transmembrane region together is also known as a scaffold.

[0311] The cap region in the chimeric pore monomer may be longer, shorter or the same length as the cap region in the pore(s) which the constriction region and / or the transmembrane region in the chimeric pore monomer is / are from or derived from. The constriction region in the chimeric pore monomer may be longer, shorter or the same length as the constriction region in the pore(s) which the cap region and / or the transmembrane region in the chimeric pore monomer is / are from or derived from. The transmembrane region in the chimeric pore monomer may be longer, shorter or the same length as the transmembrane region in the pore(s) which the cap region and / or the constriction region in the chimeric pore monomer is / are from or derived from. The constriction region in the chimeric pore monomer may be shorter than the constriction region in the pore(s) which the cap region and / or the transmembrane region in the chimeric pore monomer is / are from or derived from. Length may be measured in terms of amino acid number and / or length along the sagittal (or longitudinal) plane of the pore.

[0312] The chimeric pore monomer preferably comprises two regions. The two regions are preferably a cap region (or scaffold) and a constriction region. In such instances, the cap region (or scaffold) and the constriction region are typically from different pores.

[0313] The chimeric pore monomer preferably comprises three regions. The three regions are preferably a cap region, a constriction region, and a transmembrane region. As explained above, the cap region and transmembrane region together may also be known as a scaffold. In such instances, the constriction region may be from one pore and the cap region and thetransmembrane region (together also known as the scaffold) may from a different pore, i.e., the chimeric pore monomer is formed from / derived from two different pores. Alternatively, the cap region, the constriction region, and the transmembrane region may each be from a different pore, i.e., the chimeric pore monomer is formed from / derived from three different pores. The transmembrane region may be a transmembrane beta barrel region or a transmembrane alpha helical region. The transmembrane region is preferably a transmembrane beta barrel region.

[0314] The at least two different pores preferably comprise at least two different PorARc pores. The two different pores are preferably two different PorARc pores. The at least two or two different PorARc pores may be selected from pores comprising any one of SEQ ID NOs: 1-8 and the pores in Table 1. The at least two or two different PorARc pores may be selected from pores formed from any of SEQ ID NOs: 1-8. The at least two or two different PorARc pores may be selected from the pores in Table 1. One of the at least two different pores is preferably PorARc_Rco (pore 1 in Table 1) or PorARc_Mph (pore 2 in Table 1). One of the two different pores is preferably PorARc_Rco (pore 1 in Table 1) or PorARc_Mph (pore 2 in Table 1). The at least two different pores preferably comprise or the two different pores preferably are (a) PorARc_Rco or PorARc_Mph and (b) one of the pores in Table 1. In (b), the one of the pores in Table 1 is preferably pore 3, 8, 17, 19, 20, 25, 27 or 32. The at least two different pores preferably comprise or the two different pores preferably are PorArc_Rco and PorARc_Rco_Mph. Any reference in this paragraph to a particular pore in Table 1 (e.g., pore 1) includes any of the pores discussed above in relation to that pore Table 1 including the wild-type sequence, the sequence lacking the signal peptide and including a M at the N terminus {i.e., at position 1) and either of such pores comprising the one or more substitutions discussed above. PorARc_Rco pore preferably comprises or consists of SEQ ID NO: 1. PorARc_Mph preferably comprises or

[0315] The at least two different pores preferably comprise or the two different pores preferably are any combination shown in a row in Table 2. In relation to any row in Table 2, the cap region (or scaffold) is preferably from or derived from the pore in column A and the constriction region is preferably from or derived from the pore in column B. In relation to any row in Table 2, the constriction region is preferably from or derived from the pore in column A and the cap region (or scaffold) is preferably from or derived from the pore in column B. A reference in Table 2 to a particular pore in Table 1 {e.g., pore 3) includes any of the pores discussed above in relation to that pore in Table 1 including the wild-type sequence, the sequence lacking the signal peptide and including a M at the N terminus {i.e., at position 1) and either of such pores comprising the one or more substitutions discussed above. Where preferred pores have a sequence identifier number, these are also shown in Table 2. The pore preferably comprises or consists of the sequence identifier number.Table 2 - Preferred combinations of PorARc pores for use in the invention A B

[0316] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 1) NO: 2)

[0317] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 1) NO: 3)

[0318] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Msp (pore 4 in Table 1)

[0319] NO: 1)

[0320] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Msp (pore 5 in Table 1)

[0321] NO: 1)

[0322] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Msp (pore 6 in Table 1)

[0323] NO: 1)

[0324] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Msp (pore 7 in Table 1)

[0325] NO: 1)

[0326] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 1) NO: 4)

[0327] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mpho (pore 9 in Table 1) NO: 1)

[0328] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mmu (pore 10 in Table 1) NO: 1)

[0329] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mch (pore 11 in Table 1) NO: 1)

[0330] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mho (pore 12 in Table 1) NO: 1)

[0331] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mba (pore 13 in Table 1) NO: 1)

[0332] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mch (pore 14 in Table 1) NO: 1)

[0333] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mac (pore 15 in Table 1) NO: 1)

[0334] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mtu (pore 16 in Table 1) NO: 1)

[0335] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 1) NO: 5)

[0336] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mfl (pore 18 in Table 1)

[0337] NO: 1)

[0338] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 1) NO: 6)

[0339]

[0340] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Msm (pore 20 in Table 1) NO: 1)

[0341] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Rsp (pore 21 in Table 1) NO: 1)

[0342] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Rsp (pore 22 in Table 1) NO: 1)

[0343] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Rsp (pore 23 in Table 1) NO: 1)

[0344] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Rsp (pore 24 in Table 1) NO: 1)

[0345] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 1) NO: 7)

[0346] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Rsp (pore 26 in Table 1) NO: 1)

[0347] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 1) NO: 8)

[0348] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Rsp (pore 28 in Table 1) NO: 1)

[0349] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Rtr (pore 29 in Table 1) NO: 1)

[0350] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Nma (pore 30 in Table 1) NO: 1)

[0351] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Nsp (pore 31 in Table 1) NO: 1)

[0352] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Aku (pore 32 in Table 1) NO: 1)

[0353] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Ncy (pore 33 in Table 1) NO: 1)

[0354] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Nte (pore 34 in Table 1) NO: 1)

[0355] PorARc_Gcr (pore 35 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0356] PorARc_Rpy (pore 36 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0357] PorARc_Rpy (pore 37 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0358] PorARc_Aku (pore 38 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0359]

[0360] PorARc_Nha (pore 39 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0361] PorARc_Nfl (pore 40 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0362] PorARc_Rsp14 (pore 41 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0363] PorARc_Rsp_BP_241 (pore 26 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0364] PorARc_Mfl (pore 42 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0365] PorARc_Mba (pore 43 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0366] PorARc_RruB (pore 44 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0367] PorARc_Mwo (pore 45 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0368] PorARc_Mma (pore 46 in Table 1) PorARc_Mph (pore 2 in Table 1 or SEQ ID NO: 2)

[0369] PorARc_Rco (pore 1 in Table 1 or SEQ ID PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 1) NO: 3)

[0370] PorARc_Gcr (pore 35 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0371] PorARc_Rpy (pore 36 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0372] PorARc_Rpy (pore 37 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0373] PorARc_Aku (pore 38 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0374] PorARc_Nha (pore 39 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0375] PorARc_Nfl (pore 40 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0376] PorARc_Rsp14 (pore 41 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0377] PorARc_Rsp_BP_241 (pore 26 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0378] PorARc_Mfl (pore 42 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0379]

[0380] PorARc_Mba (pore 43 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0381] PorAR.c_R.ruB (pore 44 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0382] PorARc_Mwo (pore 45 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0383] PorARc_Mma (pore 46 in Table 1) PorARc_Msp (pore 3 in Table 1 or SEQ ID NO: 3)

[0384] PorARc_Gcr (pore 35 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0385] PorARc_Rpy (pore 36 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0386] PorARc_Rpy (pore 37 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0387] PorARc_Aku (pore 38 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0388] PorARc_Nha (pore 39 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0389] PorARc_Nfl (pore 40 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0390] PorARc_Rsp14 (pore 41 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0391] PorARc_Rsp_BP_241 (pore 26 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0392] PorARc_Mfl (pore 42 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0393] PorARc_Mba (pore 43 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0394] PorARc_RruB (pore 44 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0395] PorARc_Mwo (pore 45 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0396] PorARc_Mma (pore 46 in Table 1) PorARc_Mrh (pore 8 in Table 1 or SEQ ID NO: 4)

[0397] PorARc_Gcr (pore 35 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 4)

[0398] PorARc_Rpy (pore 36 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 4)

[0399]

[0400] PorARc_Rpy (pore 37 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0401] PorARc_Aku (pore 38 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0402] PorARc_Nha (pore 39 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0403] PorARc_Nfl (pore 40 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0404] PorARc_Rsp14 (pore 41 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0405] PorARc_Rsp_BP_241 (pore 26 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0406] PorARc_Mfl (pore 42 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0407] PorARc_Mba (pore 43 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0408] PorARc_RruB (pore 44 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0409] PorARc_Mwo (pore 45 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0410] PorARc_Mma (pore 46 in Table 1) PorARc_Mel (pore 17 in Table 1 or SEQ ID NO: 5)

[0411] PorARc_Gcr (pore 35 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0412] PorARc_Rpy (pore 36 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0413] PorARc_Rpy (pore 37 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0414] PorARc_Aku (pore 38 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0415] PorARc_Nha (pore 39 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0416] PorARc_Nfl (pore 40 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0417] PorARc_Rsp14 (pore 41 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0418] PorARc_Rsp_BP_241 (pore 26 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0419]

[0420] PorARc_Mfl (pore 42 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0421] PorARc_Mba (pore 43 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0422] PorARc_RruB (pore 44 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0423] PorARc_Mwo (pore 45 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0424] PorARc_Mma (pore 46 in Table 1) PorARc_Mco (pore 19 in Table 1 or SEQ ID NO: 6)

[0425] PorARc_Gcr (pore 35 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Rpy (pore 36 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Rpy (pore 37 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Aku (pore 38 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Nha (pore 39 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Nfl (pore 40 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Rsp14 (pore 41 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Rsp_BP_241 (pore 26 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Mfl (pore 42 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Mba (pore 43 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_RruB (pore 44 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Mwo (pore 45 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Mma (pore 46 in Table 1) PorARc_Msm (pore 20 in Table 1) PorARc_Gcr (pore 35 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0426] PorARc_Rpy (pore 36 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0427] PorARc_Rpy (pore 37 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0428] PorARc_Aku (pore 38 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0429] PorARc_Nha (pore 39 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0430] PorARc_Nfl (pore 40 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0431] PorARc_Rsp14 (pore 41 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0432]

[0433] PorARc_Rsp_BP_241 (pore 26 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0434] PorARc_Mfl (pore 42 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0435] PorARc_Mba (pore 43 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0436] PorARc_RruB (pore 44 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0437] PorARc_Mwo (pore 45 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0438] PorARc_Mma (pore 46 in Table 1) PorARc_Rsp (pore 25 in Table 1 or SEQ ID NO: 7)

[0439] PorARc_Gcr (pore 35 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0440] PorARc_Rpy (pore 36 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0441] PorARc_Rpy (pore 37 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0442] PorARc_Aku (pore 38 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0443] PorARc_Nha (pore 39 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0444] PorARc_Nfl (pore 40 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0445] PorARc_Rsp14 (pore 41 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0446] PorARc_Rsp_BP_241 (pore 26 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0447] PorARc_Mfl (pore 42 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0448] PorARc_Mba (pore 43 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0449] PorARc_RruB (pore 44 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0450] PorARc_Mwo (pore 45 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0451] PorARc_Mma (pore 46 in Table 1) PorARc_Rsp (pore 27 in Table 1 or SEQ ID NO: 8)

[0452]

[0453] The at least two different pores may be MspA and a PorARc pore. The MspA may be any of those described above, including with reference to SEQ ID NO: 11 and 12. The PorARc pore may be any of those described above, including with reference to SEQ ID NOs: 1-8 and Table 1.

[0454] Preferred chimeric pore monomers for use in the invention are shown in SEQ ID NOs: 3-49 in WO 2024 / 089270 (incorporated by reference herein in its entirety).

[0455] Stabilisation and other mutations

[0456] The pore monomer or the modified pore monomer preferably comprises one or more modifications which stabilise a pore formed from the monomer and / or improve the ability of a pore formed from the monomer to characterise a target analyte. The cap region (or scaffold) and / or the constriction region preferably comprise one or more modifications which stabilise a pore formed from the monomer and / or improve the ability of a pore formed from the monomer to characterise a target analyte. One or more of, such as all of, the cap region, the constriction region, and the transmembrane region preferably comprise one or more modifications which stabilise a pore formed from the monomer and / or improve the ability of a pore formed from the monomer to characterise a target analyte.

[0457] The one or more modifications are preferably (a) one or more deletions, (b) one or more substitutions, (c) one or more additions or (d) any combination of (a) to (c), including (a) and (b), (a) and (c), (b) or (d) or (a), (b) and (c). The one or more modifications are preferably one or more substitutions. The one or more modifications are preferably one or more of the substitutions to PorARc discussed above, especially in relation to Table 1, or MspA discussed above. Suitable one or more modifications are known in the art, including from the various patent applications cited above.

[0458] Increasing the distance between the surface region and the constriction region

[0459] The pore monomer is modified to increase the distance between the surface region and the constriction region. The pore monomer may be elongated to increase the distance between the surface region and the constriction region. The length (such as the vertical length) of the pore monomer may be increased to increase the distance between the surface region and the constriction region. The skilled person is capable of determining whether or not a pore monomer has been modified or elongated in accordance with the invention. For instance, the skilled person can compare the modified pore monomer with the natural or wild-type sequence, such as SEQ ID NO: 1.In all embodiments, the modified pore monomer is modified or elongated compared with the unmodified pore monomer. The modified pore monomer is modified or elongated compared with the pore monomer without of any of the modifications or elongations of the invention discussed below. The modified pore monomer is modified or elongated compared with the pore monomer lacking any of the insertions or additions of amino acid(s) discussed in more detail below. Similarly, in all embodiments, the distance between the surface region and the constriction region is increased compared with the unmodified pore monomer. The distance between the surface region and the constriction region is increased compared with the pore monomer without of any of the modifications or elongations of the invention discussed below. The distance between the surface region and the constriction region is increased compared with the pore monomer lacking any of the insertions or additions of amino acid(s) discussed in more detail below. Unmodified pore monomers that may be used in the invention and the distances between their surface regions and constriction regions are discussed in detail above. The unmodified pore monomer or the pore monomer without or lacking any of the modifications, elongations, insertions or additions of the invention may be a wild-type or native pore monomer or may be a pore monomer comprising other modifications or mutations not directly related to the invention. Such modifications are discussed above and below.

[0460] The distance between the surface region and the constriction region may be increased by any amount. For instance, the distance may be increased by at least about 2%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 43.5%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 69.7%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97.1%, at least about 100%, at least about 110%, at least about 113%, at least about 120%, at least about 130%, at least about 140%, at least about 149%, or at least about 150%. The at least one property is preferably increased by at least about 1.1 fold, at least about 1.49 fold, at least about 1.5 fold, at least about 2-fold, such as by at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, or at least about 10-fold.

[0461] The distance between the surface region and the constriction region is preferably increased by about 4 Å or more. The distance is preferably increased by from about 4 Å to about 100 Å, such as from about 4.5 Å to about 95 Å, from about 5 Å to about 92 Å, from about 5.5 Å to about 90 Å, from about 6 Å to about 80 Å, from about 7 Å to about 70 Å, from about 8 Å to about 60 Å, from about 9 Å to about 50 Å or from about 10 Å to about 40 Å.The distance may be increased by about 4 Å, about 4.5 Å, about 5 Å, about 5.5 Å, about 6 Å, about 6.5 Å, about 7 Å, about 7.5 Å, about 8 Å, about 8.5 Å, about 9 Å, about 9.5 Å, about 10 Å, about 10.5 Å, about 10.8 Å, about 11 Å, about 11.5 Å, about 12 Å, about 12.5 Å, about 13 Å, about 13.5 Å, about 14 Å, about 14.5 Å, about 15 Å, about 15.5 Å, about 16 Å, about 16.5 Å, about 17 Å, about 17.3 Å, about 17.5 Å, about 18 Å, about 18.5 Å, about 19 Å, about 19.5 Å, about 20 Å, about 20.5 Å, about 21 Å, about 21.5 Å, about 22 Å, about 22.5 Å, about 23 Å, about 23.5 Å, about 24 Å, about 24.1 Å, about 24.5 Å, about 25 Å, about 25.5 Å, about 26 Å, about 26.5 Å, about 27 Å, about 27.5 Å, about 28 Å, about 28.1 Å, about 28.5 Å, about 29 Å, about 29.5 Å, about 30 Å, about 30.5 Å, about 31 Å, about 31.5 Å, about 32 Å, about 32.5 Å, about 33 Å, about 33.5 Å, about 34 Å, about 34.5 Å, about 35 Å, about 35.5 Å, about 36 Å, about 36.5 Å, about 37 Å, about 37.1 Å, about 37.5 Å, about 38 Å, about 38.5 Å, about 39 Å, about 39.5 Å, about 40 Å, about 40.5 Å, about 41 Å, about 41.5 Å, about 42 Å, about 42.5 Å, about 43 Å, about 43.5 Å, about 44 Å, about 44.5 Å, about 45 Å, about 45.5 Å, about 46 Å, about 46.5 Å, about 47 Å, about 47.5 Å, about 48 Å, about 48.5 Å, about 49 Å, about 49.5 Å, about 50 Å, about 50.5 Å, about 51 Å, about 51.5 Å, about 52 Å, about 52.5 Å, about 53 Å, about 53.5 Å, about 54 Å, about 54.5 Å, about 55 Å, about 55.5 Å, about 56 Å, about 56.5 Å, about 57 Å, about 57.5 Å, about 58 Å, about 58.5 Å, about 59 Å, about 59.5 Å, about 60 Å, about 60.5 Å, about 61 Å, about 61.5 Å, about 62 Å, about 62.5 Å, about 63 Å, about 63.5 Å, about 64 Å, about 64.5 Å, about 65 Å, about 65.5 Å, about 66 Å, about 66.5 Å, about 67 Å, about 67.5 Å, about 68 Å, about 68.5 Å, about 69 Å, about 69.5 Å, about 70 Å, about 70.5 Å, about 71 Å, about 71.5 Å, about 72 Å, about 72.5 Å, about 73 Å, about 73.5 Å, about 74 Å, about 74.5 Å, about 75 Å, about 75.5 Å, about 76 Å, about 76.5 Å, about 77 Å, about 77.5 Å, about 78 Å, about 78.5 Å, about 79 Å, about 79.5 Å, about 80 Å, about 80.5 Å, about 81 Å, about 81.5 Å, about 82 Å, about 82.5 Å, about 83 Å, about 83.5 Å, about 84 Å, about 84.5 Å, about 85 Å, about 85.5 Å, about 86 Å, about 86.5 Å, about 87 Å, about 87.5 Å, about 88 Å, about 88.5 Å, about 89 Å, about 89.5 Å, about 90 Å, about 90.5 Å, about 91 Å, about 91.5 Å, about 92 Å, about 92.5 Å, about 93 Å, about 93.5 Å, about 94 Å, about 94.5 Å, about 95 Å, about 95.5 Å, about 96 Å, about 96.5 Å, about 97 Å, about 97.5 Å, about 98 Å, about 98.5 Å, about 99 Å, about 99.5 Å, or about 100 Å. The distance may be increased by about 41.7 Å or by about 48.3 Å. These distances are compared with pore monomer that has not been modified in accordance with the invention.

[0462] The distance between the surface region and the constriction region is preferably increased by the length equivalent to about 1 or more nucleotides. The distance is preferably increased by the length equivalent to from about 1 nucleotide to about 50 nucleotides, such as from about 2 nucleotides to about 45 nucleotides, from about 3 nucleotides to about 40 nucleotides, from about 4 nucleotides to about 35 nucleotides or from about 5 nucleotides to about 30 nucleotides. The distance may be increased by the length equivalent to about 1nucleotide, about 2 nucleotides, about 3 nucleotides, about 4 nucleotides, about 5 nucleotides, about 6 nucleotides, about 7 nucleotides, about 8 nucleotides, about 9 nucleotides, about 10 nucleotides, about 11 nucleotides, about 12 nucleotides, about 13 nucleotides, about 14 nucleotides, about 15 nucleotides, about 16 nucleotides, about 17 nucleotides, about 18 nucleotides, about 19 nucleotides, about 20 nucleotides, about 21 nucleotides, about 22 nucleotides, about 23 nucleotides, about 24 nucleotides, about 25 nucleotides, about 26 nucleotides, about 27 nucleotides, about 28 nucleotides, about 29 nucleotides, about 30 nucleotides, about 31 nucleotides, about 32 nucleotides, about 33 nucleotides, about 34 nucleotides, about 35 nucleotides, about 36 nucleotides, about 37 nucleotides, about 38 nucleotides, about 39 nucleotides, about 40 nucleotides, about 41 nucleotides, about 42 nucleotides, about 43 nucleotides, about 44 nucleotides, about 45 nucleotides, about 46 nucleotides, about 47 nucleotides, about 48 nucleotides, about 49 nucleotides, or about 50 nucleotides. These distances relate to the number of nucleotides equivalent to the length between and including the surface region and the constriction region when a polynucleotide analyte passes through a pore or pore complex formed from the modified pore monomer. These distances are compared with pore monomer that has not been modified in accordance with the invention.

[0463] The distance between the surface region and the constriction region is preferably increased by the length equivalent to about 1 or more nucleotides. The distance is preferably increased by the length equivalent to from about 1 nucleotide to about 50 amino acids, such as from about 2 amino acids to about 45 amino acids, from about 3 amino acids to about 40 amino acids, from about 4 amino acids to about 35 amino acids or from about 5 amino acids to about 30 amino acids. The distance may be increased by the length equivalent to about 1 nucleotide, about 2 amino acids, about 3 amino acids, about 4 amino acids, about 5 amino acids, about 6 amino acids, about 7 amino acids, about 8 amino acids, about 9 amino acids, about 10 amino acids, about 11 amino acids, about 12 amino acids, about 13 amino acids, about 14 amino acids, about 15 amino acids, about 16 amino acids, about 17 amino acids, about 18 amino acids, about 19 amino acids, about 20 amino acids, about 21 amino acids, about 22 amino acids, about 23 amino acids, about 24 amino acids, about 25 amino acids, about 26 amino acids, about 27 amino acids, about 28 amino acids, about 29 amino acids, about 30 amino acids, about 31 amino acids, about 32 amino acids, about 33 amino acids, about 34 amino acids, about 35 amino acids, about 36 amino acids, about 37 amino acids, about 38 amino acids, about 39 amino acids, about 40 amino acids, about 41 amino acids, about 42 amino acids, about 43 amino acids, about 44 amino acids, about 45 amino acids, about 46 amino acids, about 47 amino acids, about 48 amino acids, about 49 amino acids, or about 50 amino acids. These distances relate to the number of amino acids equivalent to the length between and including the surface region and the constriction region when a polypeptide analyte passes through a pore or pore complex formed from themodified pore monomer. These distances are compared with pore monomer that has not been modified in accordance with the invention.

[0464] In modified PorARc pore monomers or modified pore MspA pore monomers, the distance between the surface region and the constriction region is preferably increased to at least 79 Å. The distance is preferably increased to from about 79 Å to about 200 Å, such as from about 80 Å to about 180 Å, from about 90 Å to about 160 Å, or from about 100 Å to about 200 Å. The distance may be increased to about or at least about 79 Å, about or at least about 80 Å, about or at least about 81 Å, about or at least about 82 Å, about or at least about 83 Å, about or at least about 84 Å, about or at least about 85 Å, about or at least about 86 Å, about or at least about 87 Å, about or at least about 88 Å, about or at least about 89 Å, about or at least about 90 Å, about or at least about 91 Å, about or at least about 92 Å, about or at least about 93 Å, about or at least about 94 Å, about or at least about 95 Å, about or at least about 96 Å, about or at least about 97 Å, about or at least about 98 Å, about or at least about 99 Å, about or at least about 100 Å, about or at least about 101 Å, about or at least about 102 Å, about or at least about 103 Å, about or at least about 104 Å, about or at least about 105 Å, about or at least about 106 Å, about or at least about 107 Å, about or at least about 108 Å, about or at least about 109 Å, about or at least about 110 Å, about or at least about 111 Å, about or at least about 112 Å, about or at least about 113 Å, about or at least about 114 Å, about or at least about 115 Å, about or at least about 116 Å, about or at least about 117 Å, about or at least about 118 Å, about or at least about 119 Å, about or at least about 120 Å, about or at least about 121 Å, about or at least about 122 Å, about or at least about 123 Å, about or at least about 124 Å, about or at least about 125 Å, about or at least about 126 Å, about or at least about 127 Å, about or at least about 128 Å, about or at least about 129 Å, about or at least about 130 Å, about or at least about 131 Å, about or at least about 132 Å, about or at least about 133 Å, about or at least about 134 Å, about or at least about 135 Å, about or at least about 136 Å, about or at least about 137 Å, about or at least about 138 Å, about or at least about 139 Å, about or at least about 140 Å, about or at least about 141 Å, about or at least about 142 Å, about or at least about 143 Å, about or at least about 144 Å, about or at least about 145 Å, about or at least about 146 Å, about or at least about 147 Å, about or at least about 148 Å, about or at least about 149 Å, about or at least about 150 Å, about or at least about 151 Å, about or at least about 152 Å, about or at least about 153 Å, about or at least about 154 Å, about or at least about 155 Å, about or at least about 156 Å, about or at least about 157 Å, about or at least about 158 Å, about or at least about 159 Å, about or at least about 160 Å, about or at least about 161 Å, about or at least about 162 Å, about or at least about 163 Å, about or at least about 164 Å, about or at least about 165 Å, about or at least about 166 Å, about or at least about 167 Å, about or at least about 168 Å, about or at least about 169 Å, about or at least about 170 Å, about or at least about 171 Å, about or at least about 172 Å, about or at leastabout 173 Å, about or at least about 174 Å, about or at least about 175 Å, about or at least about 176 Å, about or at least about 177 Å, about or at least about 178 Å, about or at least about 179 Å, about or at least about 180 Å, about or at least about 181 Å, about or at least about 182 Å, about or at least about 183 Å, about or at least about 184 Å, about or at least about 185 Å, about or at least about 186 Å, about or at least about 187 Å, about or at least about 188 Å, about or at least about 189 Å, about or at least about 190 Å, about or at least about 191 Å, about or at least about 192 Å, about or at least about 193 Å, about or at least about 194 Å, about or at least about 195 Å, about or at least about 196 Å, about or at least about 197 Å, about or at least about 198 Å, about or at least about 199 Å or about or at least about 200 Å.

[0465] The channel forming region and / or the transmembrane region in the pore monomer or the modified pore monomer may comprise or consist of a beta-sheet and / or one or more alpha helices. The channel in the pore formed from the pore monomer or the modified pore monomer may comprise or consist of a beta-barrel and / or an alpha-helix barrel. The channel may comprise or consist of a beta-barrel. The channel may comprise or consist of an alpha-helix barrel. The channel may comprise or consist of a beta-barrel and an alphahelix barrel.

[0466] The pore monomer or the modified pore monomer may comprise a beta-sheet and / or one or more alpha helices. The pore monomer or the modified pore monomer may comprise or consist of a beta-sheet. The pore monomer or the modified pore monomer may comprise or consist of one or more alpha helices. The pore monomer or the modified pore monomer may comprise or consist of a beta-sheet and one or more alpha helices.

[0467] The pore monomer or modified pore monomer typically comprises one or more betastrands. The transmembrane region typically comprises one or more beta-strands. The beta-sheet typically comprises one or more beta-strands. The pore monomer, modified pore monomer, transmembrane region or beta-sheet may comprise any number of one or more beta-strands, such one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more or ten or more beta-strands. The pore monomer, modified pore monomer, transmembrane region or beta-sheet may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 beta-strands. The pore monomer, modified pore monomer, transmembrane region or beta-sheet may comprise 4 beta-strands or 2 beta-strands. The pore monomer, modified pore monomer, transmembrane region or beta-sheet may comprise 2 beta-strands.

[0468] The one or more alpha helices may comprise any number of one or more alpha helices, such one or more, two or more, three or more, four or more, five or more, six or more, seven ormore, eight or more, nine or more or ten or more alpha helices. The pore monomer or modified pore monomer may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alpha helices.

[0469] In some embodiments, one or more beta-strands and / or one or more alpha helices are modified to increase the distance between the surface region and the constriction region. One or more beta-strands may be modified to increase the distance between the surface region and the constriction region. One or more alpha helices may be modified to increase the distance between the surface region and the constriction region. One or more betastrands and one or more alpha helices may be modified to increase the distance between the surface region and the constriction region. The one or more beta-strands and / or one or more alpha helices are in the pore monomer or modified pore monomer.

[0470] Any number of one or more beta-strands and / or one or more alpha helices may be modified, such one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more or ten or more. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 may be modified. These numbers relate to the number of one or more beta-strands and / or one or more alpha helices modified in each modified pore monomer. Preferably, all of the one or more beta-strands and / or one or more alpha helices.

[0471] Any number of one or more beta-strands may be modified, such one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more or ten or more beta-strands. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 beta-strands may be modified. Preferably, all of the one or more beta-strands are modified. All 4 beta-strands may be modified. Both of the 2 beta-strands may be modified.

[0472] The one or more beta-strands and / or one or more alpha helices are typically elongated. In any of the paragraphs above discussing modifying one or more beta-strands and / or one or more alpha helices, the term "modified" may be replaced with "elongated". Specific methods of doing this are discussed in more detail below.

[0473] When two or more beta-strands and / or one or more alpha helices are modified or elongated, one or more of the beta-strands and / or one or more alpha helices may be modified or elongated. All of the two or more beta-strands and / or two or more alpha helices may modified or elongated. The two or more beta-strands and / or two or more alpha helices may be modified or elongated to different degrees. The two or more beta-strands and / or two or more alpha helices are preferably modified or elongated to the same degree. In this context, the term "degree" typically relates to the number of amino acids added to the two or more beta-strands and / or two or more alpha helices. For instance, one amino acid may be added to one beta-strand and / or alpha helix and two amino acids may be added to a different beta-strand and / or alpha helix. Alternatively, two amino acids can be added to both beta-strands and / or alpha helices.Even if two or more beta-strands and / or one or more alpha helices are modified or elongated to the same degree, they may be modified in different ways. For instance, two amino acids may be added to one beta-strand and / or alpha helix and two different amino acids may be added to a different beta-strand and / or alpha helix. Alternatively, four consecutive amino acids may be added to one beta-strand and / or alpha helix and two separate stretches of two amino acids may be added to a different beta-strand and / or alpha helix. The identities of the four consecutive amino acids and the two separate stretches of two amino acids may be the same or different. The skilled person is capable of designing suitable modifications and elongations.

[0474] Since different beta-strands and / or alpha helices in the pore monomer may be different lengths and they may be modified or elongated to the same degree or different degrees, the two or more beta-strands and / or two or more alpha helices may be the same length or different lengths once they are modified or elongated. As discussed above, the skilled person is capable of recognising when one or more beta-strands and / or one or more alpha helices are modified or elongated, for instance with reference to the natural or wild-type sequence.

[0475] The one or more beta-strands and / or one or more alpha helices typically comprise one or more preferred positions for modifications or elongations. Each beta-strand and / or alpha helix may comprise at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9 or at least about 10 preferred positions for modifications or elongations. In any of these embodiments, preferred positions typically represent positions between amino acids in the sequence of the one or more beta-strands and / or one or more alpha helices at which one or more amino acids may be inserted. For two or more beta-sheets, two or more preferred positions typically line up approximately horizontally in the beta-sheet.

[0476] The one or more beta-strands and / or one or more alpha helices may be elongated by any amount. The one or more beta-strands and / or one or more alpha helices are preferably elongated by from about 1 to about 24 amino acids. The one or more beta-strands and / or one or more alpha helices are preferably elongated by about 1 amino acid, about 2 amino acids, about 3 amino acids, about 4 amino acids, about 5 amino acids, about 6 amino acids, about 7 amino acids, about 8 amino acids, about 9 amino acids, about 10 amino acids, about 11 amino acids, about 12 amino acids, about 13 amino acids, about 14 amino acids, about 15 amino acids, about 16 amino acids, about 17 amino acids, about 18 amino acids, about 19 amino acids, about 20 amino acids, about 21 amino acids, about 22 amino acids, about 23 amino acids or about 24 amino acids. The one or more beta-strands and / or one or more alpha helices are preferably elongated by about 2 amino acids, about 4 amino acids, about 6 amino acids, about 8 amino acids, about 10 amino acids, or about 12 amino acids.The one or more beta-strands and / or one or more alpha helices may be elongated by inserting these numbers of amino acids.

[0477] Any of the amino acid elongations / insertions disclosed herein may be consecutive or may by separated by existing sequences. For instance, if 4 amino acids are used to elongate one or more beta-strands and / or one or more alpha helices (i.e., 4 amino acids are inserted), they may be used / inserted as 4 consecutive amino acids or they may be used / inserted as 2 instances of 2 consecutive amino acids separated by existing sequences in the pore monomer.

[0478] The one or more beta-strands and / or one or more alpha helices may be elongated using any amino acid(s). The amino acid(s) may form beta-strands and / or alpha helices. The one or more beta-strands may be elongated using amino acid(s) which form one or more alpha helices. The one or more alpha helices may be elongated using amino acid(s) which form one or more beta-strands. The one or more beta-strands are preferably elongated using amino acid(s) which form one or more beta-strands. The one or more alpha helices are preferably elongated using amino acid(s) which form one or more alpha helices.

[0479] Modification or elongation of the one or more beta strands and / or one or more alpha helices typically maintains the beta strand and / or alpha helical structure of the one or more beta strands and / or one or more alpha helices. Modification or elongation of the one or more beta strands typically maintains the beta strand structure of the one or more beta strands. Modification or elongation of the one or more alpha helices typically maintains the alpha helical structure of the one or more alpha helices.

[0480] The transmembrane region or the transmembrane beta barrel region is preferably modified or elongated in accordance with the invention. The amino acid(s) are preferably located / inserted into the transmembrane region or the transmembrane beta barrel region.

[0481] The amino acid(s) may be located / inserted within of the membrane-spanning region of the transmembrane region. The one or more beta-strands are preferably elongated using amino acid(s) which form one or more beta-strands. In such embodiments, the outward facing amino acid(s) are typically hydrophobic and are located / inserted within the membranespanning region of the transmembrane region. In other embodiments, the outward facing amino acid(s) are typically polar and / or charged and are located / inserted within the membrane-spanning region of the transmembrane region.

[0482] The amino acid(s) may be located / inserted outside of the membrane-spanning region of the transmembrane region. In these embodiments, the identity / ies of the outward facing amino acid(s) is less constrained and the skilled person is capable of designing appropriate elongations / insertions.The amino(s) may be natural and / or non-natural amino acids. The amino acid(s) is / are preferably selected from tyrosine (Y), arginine (R), lysine (K), phenylalanine (F), tryptophan (W), threonine (T), valine (V), alanine (A), isoleucine (I), leucine (L), glycine (G), glutamic acid (E), aspartic acid (D), histidine (H), methionine (M), cysteine (C), proline (P), serine (S), asparagine (N,) glutamine (Q) and non-natural amino acids.

[0483] The non-natural amino acids may be referred to as chemically synthesised amino acids or unnatural amino acids. Non-natural amino acids can be synthetically prepared from their native analogs via modifications such as amine alkylation, side chain substitution, structural bond extension cyclization, and isosteric replacements within the amino acid backbone. The non-natural amino acids include, but are not limited to, hydroxyproline (Hyp), beta-alanine, citrulline (Cit), ornithine (Orn), norleucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr).

[0484] The amino acid(s) is / are preferably selected from tyrosine (Y), threonine (T), valine (V), isoleucine (I), leucine (L), glycine (G), serine (S), glutamic acid (E), lysine (K), arginine (R), asparagine (N), glutamine (Q), aspartic acid (D) and alanine (A).

[0485] For 2 amino acid elongations or insertions, the amino acids may be selected from any amino acids. The amino acids may be natural and / or non-natural amino acids. The 2 amino acid elongations or insertions are preferably selected from tyrosine (Y), arginine (R), lysine (K), phenylalanine (F), tryptophan (W), threonine (T), valine (V), alanine (A), isoleucine (I), leucine (L), glycine (G), glutamic acid (E), aspartic acid (D), histidine (H), methionine (M), cysteine (C), proline (P), serine (S), asparagine (N) and glutamine (Q). For 2 amino acid elongations or insertions, the amino acids are preferably selected from G, V, R, K, S, T, I, E, Q, D, N, H, L, A, Y, F, M, C, W and P.

[0486] For 4 amino acid elongations or insertions, the amino acids may be selected from any amino acids. The amino acids may be natural and / or non-natural amino acids. The 4 amino acid elongations or insertions are preferably selected from tyrosine (Y), arginine (R), lysine (K), phenylalanine (F), tryptophan (W), threonine (T), valine (V), alanine (A), isoleucine (I), leucine (L), glycine (G), glutamic acid (E), aspartic acid (D), histidine (H), methionine (M), cysteine (C), proline (P), serine (S), asparagine (N) and glutamine (Q). For 4 amino acid elongations or insertions, the amino acids are preferably selected from T, V, K, L, R, Y, S, I, Q, E, N, H, A, D, F, M, W, G, C, and P.

[0487] For 6 amino acid elongations or insertions, the amino acids may be selected from any amino acids. The amino acids may be natural and / or non-natural amino acids. The 6 amino acid elongations or insertions are preferably selected from tyrosine (Y), arginine (R), lysine (K), phenylalanine (F), tryptophan (W), threonine (T), valine (V), alanine (A), isoleucine (I), leucine (L), glycine (G), glutamic acid (E), aspartic acid (D), histidine (H), methionine (M),cysteine (C), proline (P), serine (S), asparagine (N) and glutamine (Q). For 6 amino acid elongations or insertions, the amino acids are preferably selected from T, V, K, S, N, I, R, L, Y, E, Q, A, D, H, F, G, M, W, P, and C.

[0488] For 8 amino acid elongations or insertions, the amino acids may be selected from any amino acids. The amino acids may be natural and / or non-natural amino acids. The 8 amino acid elongations or insertions are preferably selected from tyrosine (Y), arginine (R), lysine (K), phenylalanine (F), tryptophan (W), threonine (T), valine (V), alanine (A), isoleucine (I), leucine (L), glycine (G), glutamic acid (E), aspartic acid (D), histidine (H), methionine (M), cysteine (C), proline (P), serine (S), asparagine (N) and glutamine (Q). For 8 amino acid elongations or insertions, the amino acids are preferably selected from T, V, S, N, K, E, I, Y, R, L, Q, D, A, H, F, M, G, W, P, and C.

[0489] For 10 amino acid elongations or insertions, the amino acids may be selected from any amino acids. The amino acids may be natural and / or non-natural amino acids. The 10 amino acid elongations or insertions are preferably selected from tyrosine (Y), arginine (R), lysine (K), phenylalanine (F), tryptophan (W), threonine (T), valine (V), alanine (A), isoleucine (I), leucine (L), glycine (G), glutamic acid (E), aspartic acid (D), histidine (H), methionine (M), cysteine (C), proline (P), serine (S), asparagine (N) and glutamine (Q). For 10 amino acid elongations or insertions, the amino acids are preferably selected from T, V, S, N, E, K, I, Y, Q, R, L, D, A, H, F, G, M, W, P, and C.

[0490] For 12 amino acid elongations or insertions, the amino acids may be selected from any amino acids. The amino acids may be natural and / or non-natural amino acids. The 12 amino acid elongations or insertions are preferably selected from tyrosine (Y), arginine (R), lysine (K), phenylalanine (F), tryptophan (W), threonine (T), valine (V), alanine (A), isoleucine (I), leucine (L), glycine (G), glutamic acid (E), aspartic acid (D), histidine (H), methionine (M), cysteine (C), proline (P), serine (S), asparagine (N) and glutamine (Q). For 12 amino acid elongations or insertions, the amino acids are preferably selected from T, V, S, N, E, K, I, Y, Q, R, L, D, A, H, F, G, M, W, P, and C.

[0491] For 14 amino acid elongations or insertions, the amino acids may be selected from any amino acids. The amino acids may be natural and / or non-natural amino acids. The 14 amino acid elongations or insertions are preferably selected from tyrosine (Y), arginine (R), lysine (K), phenylalanine (F), tryptophan (W), threonine (T), valine (V), alanine (A), isoleucine (I), leucine (L), glycine (G), glutamic acid (E), aspartic acid (D), histidine (H), methionine (M), cysteine (C), proline (P), serine (S), asparagine (N) and glutamine (Q). For 14 amino acid elongations or insertions, the amino acids are preferably selected from T, V, S, N, E, K, I, Y, Q, R, L, D, A, H, F, G, M, W, P, and C.The one or more beta strands and / or one or more alpha helices may be elongated by one or more of EE, LS, FS, LE, NT, SH, CV, FD, SQ, QA, AN, AK, QD, VH, DV, LD, KL, QQ, QV, AL, RQ, Al, TI, II, HS, RI, ED, KI, YD, MV, KN, SD, HQ, KY, IG, GV, VG, GI, NL, GL, LG, NA, GA, GF, PG, GG, SA, GT, DL, NI, GY, TL, NG, NF, AG, PV, SF, GW, DF, IP, TH, QP, NR, IL, LV, RY, AE, DG, RE, RK, GH, QI, YE, NY, CS, VR, RY, LC, LI, LP, EL, IK, MW, VC, AW, PR, EP, HI, FW, HE, ML, HM, WE, MI, QG, VY, GR, YA, GM, CP, TH, LR, YL, MT, YW, MG, EM, IQ, FE, WY, PC, TS, GC, CM, MN, SM, NE, IF, TA, VP, WA, NH, ES, SL, DN, KR, MK, VV, ER, HW, LP, RS, FC, DQ, TG, NS, GD, TV, QL, KT, KM, CG, VQ, VT and LM. The one or more beta strands and / or one or more alpha helices may comprise one or more of the listed insertions.

[0492] Different combination of these two amino acid elongations / insertions may be used in the same beta strand or alpha helix or different combinations of two or more beta strands and / or two or more alpha helices. The two amino acid elongations / insertions may be used in the one or more beta strands and / or one or more alpha helices consecutively or separated by existing sequences.

[0493] The one or more beta strands and / or one or more alpha helices may be elongated by one or more of TVTV, VTVT, VKVD, NTYK, TYTN, DTKK, VLNI, VTYI, VTPG, VTVP, GVQA, NLSI, INYR, VLQL, LEYK, SQTV, LQVK, VVQV, VTYT, QQLV, VQVK, VEYE, LERI, STLL, EYKK, KVNL, LSYT, SYLL, LNYK, VMNL, LTYK, TQTF, LTQR, VYTY, LTYT, TYTL, YTVK, KTIL, LEYN, LIFK, VVTV, ITYK, TQTV, VTTK, ILTL, LTYR, TRLV, LLER, VYTL, LTFK, LTKK, VIAV, SYLA, VSQK, VWNV, LTFT, SYTF, VSRR, VVQL, LTYV, GYQY, LQNK, VETT, HTLT, SSTT, TTVK, IEKD, QDTV, SQTT, DKVR, VVRV, VDYT, SRRV, VFHK, TMNV, VINH, RRNI, HLER, EQTD, ITRK, SQTL, DTVR, VTTV, QTYT, VSVK, VVEV, HYSV, VEKK, VLRL, TKTL, LTVK, VVKI, LEYQ, SYTA, IKYK, QHNH, LNYR, TSTY, HTVR, VVRI, AEYT, SMNV, IRQK, VERT, VAKK, VENE, ITRT, SVLR, EATR, VLSI, FTYN, SYSA, FSYK, VVTL, TYTV, LTVR, VQTV, LSTT, TQTY, VTIR, LNYT, SYNF, VSYA, TYQL, LTTK, ERRV, LTDR, SREA, VRTR, ITYS, TTTV, LQQR, VERE, LTRT, TQEV, ERRR, VVNI, SMLC, IFFK, KVQV, LEYV, KTTV, VQKK, VLKV, TYTF, VSYK, VLTV, FTYT, SYNV, VATK, IVSV, TYTT, VTVK, QVSI, IEYK, ISKK, VISI, SYTL, ISYR, VITI, KQTV, ITKR, VHQH, RMEL, HQVK, LTYN, TYLA, ITTR, VEQR, QTRV, QQTQ, RQQR, VVDI, VTYS, KRTV, ISER, YEYK, SQLL, LAKK, LNRT, KMTI, YTHK, IVRV, TKTV, VRVR, VSVH, TQTL, HTRK, VTRT, LERR, TTLT, TLVR, LTYS, SYGA, VQRK, VVSV, LTRV, STTV, VSRK, VTQD, NTRV, DQVK, IYNL, TDYT, NQTV, INKK, VLTL, LTQK, AQYS, SFTL, VQRR, VLNL, LQYT, SKTV, LSYR, VLSL, LEYR, LKFK, HTYT, SYQF, VHNH, SQLQ, HSQK, IINV, YTRK, TQEL, VNTR, VQNV, YTMT, SQLV, VLRK, VTLK, IVRL, TEYT, SYTV, LRLR, VLTY, AEYS, YTEK, LEYS, VRQK, IVQV, VQVR, IALL, HYLL, LATR, VINL, FTYQ, SYNL, QLRV, LDYI, SKTT, VRER, VENI, YTRT, DDLI, IYSR, VVRL, KKLL, LLEK, VTRE, IETK, DKTV, ETVK, VYVH, SMTV, HVYR, STTA, VYML, HTFI, TKQI, LMIR, VIQN, VQYN, SLTY, NQTK, LEYT, SKTL, IKEK, IISN, TYEL, NSTK, IEQV, IQRQ, TETI, VQQK, NVNI, LNYS, AQTL, INIK, TWHL, QEYR, EYLL, LHTK, VVSL, SYQL, LRYR, TLRV, SYTI, LYEK, VIEV, VLYR, VIRV, FEYQ, VAVK, VIQI, NYLV, IQYK, VVNF, SMTL, FSQK, VYSY, NQTL, YSYR, VNIH,LIRN, HIER, VSTD, TQTR, DVQR, LTEK, LVQI, LTYQ, KQQV, IQQR, VLTI, QTYK, ITQR, VLKL, AEYV, LKKK, RLSV, SYSL, VYRD, AYNR, DQKR, VLSY, SKNV, YSVR, VESE, VDRT, KSTS, ESTR, AEYK, KEQI, LEKT, KYTL, IEVK, FEYK, AMTL, LSVK, VEYK, VTYK, VTYR, VLNV, ITVR, ILNL, TQNV, LLTK, VEAT, HTRT, SDSS, TATK, VDKH, TERK, HKIK, VNED, VKTV, SQTK, DAQK, RIQI, ILTF, FTYK, IISV, LTYI, QTTL, VTKR, SISL, VEYN, SQSV, LSSK, TINV, FLYN, SQLI, VFKK, IIKV, SYLG, VKYR, IEQE, YTYK, EQNK, IQNK, NVNV, VNVR, TVSV, FQYV, SYLM, INRK, LEYI, TQKV, VMVR, VIQV, LKYT, SYQV, VLQK, VYSL, INYT, SYLV, LYRK, VLNN, TRTV, NTTR, KVNV, VIVV, VKYT, SRTI, VYEK, LTSR, TRTF, VTER, VTQI, AEYI, IQTR, VSTH, ITTK, SRLT, HTKR, VSEI, IDTT, IQKK, TQLY, ILQR, SLNL, SKNL, LALR, IERE, HERK, TELK, ELKK, VENL, QRYT, SKST, LNEK, LDYT, IRRK, VINV, YEYR, EQTV, VNRR, IYRN, SKLL, NHRK, TIQL, IERI, STTL, LSNK, VLEV, LRYV, TYEA, VIVK, VQNN, HTYK, TRTY, NQQK, VSYR, RVWV, IEYV, VYSK, INNN, LNKV, VQAQ, TNYT, TSTQ, QTKK, YAKK, IIEV, TYTA, TIYI, STNV, IYTK, TVTH, SQTY, ITDK, VINI, ITWR, IQNI, QTYI, IMRK, IEQA, VTRK, SYQS, AQNR, LNYV, VTVR, AYEV, VIEI, ATYR, VQKH, LTQT, TTEA, HKEK, IKKD, NEYK, SKTN, VHNV, LDRK, VNEK, INRH, LEMT, SQTQ, HRSK, QINV, SYNA, VKQR, IEYT, YYNV, ILVK, SKLV, VSKK, VVSI, ISYK, SMTF, ISVK, TTLV, KVSV, ITYT, SLTV, VESV, YNLT, RRTI, VTQK, THTV, LTKR, IVRI, WEYK, KITI, NMTI, ITRR, SYDL, TDIR, TRSL, RIER, IVSI, FNYK, INKT, VIRY, YARR, ILSI, ISVR, TMTV, VTSR, INKR, VAIT, VEYR, ERTV, TIEK, INAL, LERK, STTI, LAER, HTQR, YQYT, SRLV, LQVR, VSRS, SRSK, VEYT, SQMV, LRIR, QTRI, TNTD, TTSK, DNLR, SMSQ, KVKV, SKTI, VTEK, TVYV, SYEL, VYWK, EYNA, VKSV, VEYV, STSV, VSEK, VANV, YEYT, TNLF, VQQR, VLYL, RQAV, VVNV, VNQR, VKTH, LTTA, SRTA, HSSR, VLTT, TTYV, TKTT, TTEK, VTNL, LTYE, SQSA, LSQR, VTYV, VSTR, VVIV, VLQR, KALL, SQLA, QDIT, RQLN, ELKE, VNQH, RNYT, SRLD, HQER, VVQI, SMLL, LSYS, SYTG, ISWK, ISSI, YERT, HQLL, ISSR, IVNV, QKYI, VFQR, VISV, LTYC, VYTF, FEYT, SYSF, FTKK, INVK, VQTH, LDYR, HTSR, VTRH, IERV, HAVR, TLSL, TVDI, IDYT, VHSH, VTEI, EVTI, ITVK, SLTA, VRQE, LTET, SRLF, TYNL, HNYT, SKSI, LNQR, IVNI, QKNV, VSVR, VYIL, TYGL, LIYR, KICI, YNYI, SQNY, MNYT, ILKK, IENH, LSRT, SYTY, HSKK, VIHK, KHRK, TVRV, VRTK, VIHI, ISYI, IHWK, IYRY, LDTQ, YRQK, VLVL, LTLK, KVLE, IEYI, EQKR, VFKI, TQEN, ITQK, TITI, VNYN, TVNL, NTYV, TQTN, LSEK, VTNV, YNYS, TREA, VRRR, VVTI, ISQD, TQTQ, DQKA, ITTL, ATYN, VLRD, SMLV, DKYK, VYNY, RSYV, KQTD, YTLR, VYNV, STLV, VNKK, TLTL, RVSV, VSQR, FSYT, SMTI, ISTR, LTWV, TGSV, VTRR, VLSV, RKTV, NLTI, LNYN, LTKT, NQNF, ESKR, IQKQ, SKNE, QQVK, VQSQ, NTYT, SQQT, QSTR, VEEV, LTRR, SQAT, VLVR, TVNI, NYLL, SQLH, LNKK, IYYR, LRER, VQEV, YTRI, KQLI, VAQS, ITYQ, TINT, HTYL, TQNY, TYTK, SEYR, SQSG, VQEK, VIRL, KQSV, IENI, LTRK, IKVK, TQTT, TTNK, VKRL, TKEL, LRQR, NTYN, SYTT, IYVK, TQNL, VYQY, DTQV, YQVR, VQSN, AKTV, NSQR, QVNI, INQR, VETE, ETKK, VLML, RQQI, LMTK, TVSI, VNTN, NTVK, TYTY, VETI, ITEK, KKTV, ITES, QWNL, LSYK, TLLV, AYTL, VAYK, VLWR, SYVA, VHKK, KYRI, VDSV, VFNT, YTYQ, SANY, TTKK, VLYK, KVQI, SGTV, ITKK, VAYR, TYQA, YRYR, HNYK, TSTA, VNVK, IDED, IKRI, RLVL, KTTA, LEEK, IIQV, SQQY, YSKK, TASL, YTYR, LERT, SQTA, VTSK, LETR, YAYR, VQYR, LVHV, LEYG, VYYK, VVNT, QMTV, TTIK, LNVR, TWKE, EKTK, LTVT, NTRT,NQTN, VLRY, YSYK, SNMF, ISIR, VISH, HSVK, VRKA, VYSH, FTYV, HSYK, TMTA, NQLK, TMTT, VGTL, WTQK, VAMA, SEYV, RKLN, AEER, IENL, FTRK, SQTF, VITV, FTYR, VSEE, TRSV, LNKR, VTRI, VWKH, QNYT, AQTF, VNAR, SQTI, ISTK, VSTL, EKTV, LTER, KLSL, LSTR, HLSL, QDYT, SYSN, VISK, KTVK, IFHL, LHVK, VEVR, YTYT, VFTI, STYT, ISQK, QVNV, SQSI, VKRK, IQRT, LETK, IHME, VERK, EMTR, VVSH, RESH, QDRT, VVEL, LLYT, SFTV, LSQK, VEKH, EYLF, HKYK, TVNV, TIQV, QTVT, ITTV, QNHT, KSTT, VTDK, VLDI, TYNM, ITYR, QRSQ, TKTY, IVKV, SMLF, VKKK, AQYK, SRTT, IHEK, KVAL, YEYL, SQNV, LYFK, IEHE, SLTE, EHVR, TVHV, ARNV, VHVR, TVAL, LAYK, ITSK, LSKK, TLEN, QQYT, NTKK, FQQK, VNYK, VLER, VDTN, QTNK, SQTD, NSVK, ISIK, QVDI, AQTV, ISKR, INQH, LEGV, KQTF, HVQK, IVSL, QQYR, LKYR, ILNI, ITYV, AQTI, IIVK, VRRE, TRNL, ERER, VVHL, SRSA, LAQK, KVLI, QVYV, RQLL, VYRK, IEKQ, HQNV, ELKL, LKYK, ATRT, VLQS, SHNL, SQVK, IETT, KQTY, VTKK, VTSN, ATYT, SSTQ, NSTR, VAIL, KKTL, LSER, AQNV, VEER, FTRR, SYLY, REER, LETT, IFYK, IEEE, VTRQ, SKTY, EEQK, LKQK, LTTR, SRLI, VMDR, IATL, LPQK, VDRN, TERT, VNTT, TSYK, TLNV, ISFK, LDYK, VVTY, YTKR, ILRL, YTYI, LSVR, HYTL, VQTN, SQTS, INTV, KQTL, VTHK, RLSA, ANYN, SYDV, ASFR, IYSV, TQTI, IESE, HTTT, SQLN, ESKK, AYTV, YRVR, KVNI, LDYV, EIRV, SREV, KQVE, TRTT, ETVR, TVHF, FYKK, VNYQ, KQNV, ILTR, SYSV, TYQF, YTTK, KEYT, TNLD, NYKK, VSAH, VYER, VLRI, FDYR, TFLF, ILTK, VEET, QERK, QQKT, TQTK, VQED, SYTK, DAKK, LSSR, VLEN, YYTF, NSKK, KENH, DERV, SQTH, HNEK, LTYF, TYTI, VQIK, VTTS, TQEF, SSVR, SYSY, VTFR, TRTL, VYKW, TREV, WKTK, RLRL, ISYT, TQLQ, LLKK, VQVV, QERT, SQLT, VFSR, EYLV, VQYK, TTYT, VSIK, TFEY, EYTL, EMTI, IQRR, VNNH, SYTE, HSLK, HTYV, SYTS, ISRK, VETR, RTVR, TLQL, IITV, DRLV, VSWR, IVNH, HTTI, VRSD, NETS, DSRK, IVDV, YTYA, VFAL, LAVK, LNRE, HSEK, VEAK, INRT, NRSE, KAVK, QISV, VYTK, SYSG, VNQK, VNRK, RIHF, IDYV, FNKR, VERR, KRLD, RREK, VVTQ, TTYK, TRTD, QTER, IIRI, ILKR, GYVV, VRYK, TITY, TQLF, VLSF, FSFK, EIDI, SQAV, IAQR, TINI, INRR, VIRI, IRYK, WSYR, LQYV, IQYR, VVQT, DTYT, TQVR, DYSV, LLYR, LLNL, FSYS, RQTL, ISEK, ILQH, HLTR, VHAH, HSHR, WMV, NMML, VMSK, SRTF, TNYR, SQQV, VTSD, SQST, DSKK, SMTA, LQQK, TVTI, ITRD, VDKR, AQRV, DYQK, FEYR, SMTY, YSEK, VEQI, YNFT, SMLM, TVTF, VGVG, GVNF, IEVE, VEVG, ISVG, IGVG, VTVG, VSVG, ITVT, VEVE, VNVG, LGVG, VDVD, GVNI, IEVG, GVNG, LEVE, VSVS, LSVG, LEVG, VTVV, VSVT, VDVV, IDVD, VGIG, ESVS, VEVV, KEVG, VQVG, ITVG, VSVN, VAVV, VEIG, LRVG, LQVG, ISVS, GVGF, VAVA, IGIG, GKNF, LEVT, LDVD, VQVQ, VTVI, IAVA, VNVD, INVN, IIVI, RSVG, GYNI, IQVQ, VNVN, LSVS, GLNF, ISVT, IVVV, VTVE, LTVG, VNVT, IEVN, AEVG, GEVG, VVVV, VSVE, ISVL, AGVG, INEN, GVTF, VHVH, IEVV, VSIG, LGFG, VVQY, VNSN, KVRI, HEYA, IEMR, PVTV, HVQV, LEMV, VNNY, NTKI, VLLL, TSTE, LTDT, VVLN, HERR, VFTF, TLYL, FQYT, VNYS, VVDV, IIYT, VLRF, VVVH, TISV, EVNV, KVRV, LEYD, EVSV, IVTI, KETV, IISL, IRYT, RENV, LNYI, VLEL, IETH, LDRR, EEHV, QYQY, INYN, VITH, VQQN, IINY, ITRV, VVAI, RIAS, VVYT, EVKV, IHDV, TITV, TEYK, VVVL, VVHV, TVQI, TYVV, LTQN, SIYI, DARN, VMQH, LERS, VWEV, EYTY, YTYN, VFTL, LLRY, FEYD, VFQH, VISL, VQYV, HEYR, VVTT, ATYV, VKKI, QTYQ, VIDV, VDQH, IERK, EVTV, ILRT, FEYV, VYDH, VNTS, ITNR, RINI, IQSD, YERR, MIQI, ITTT, IYTY, ILVL, LVYT, EITI, KTNA, LTRE, RVNV, LTYL, VFSI, VSEV,YTET, VVKV, TNTN, NTNT, YTGG, ATGT, RTRT, ERTR, GTGS, TGTL, EGTF, GTLK, TLTG, LTGT, TDEI, TLKG, TDVI, TNTL, NGLT, GTYT, TGTQ, RENS, TSGE, EGKN, HTES, TRTN, LTGE, YEVG, VTGS, LNQK, VTYN, STST, TSTS, VTYQ, GVDV, DYGL, VDYK, VDYN and RVDV. The one or more beta strands and / or one or more alpha helices may comprise one or more of the listed insertions. Different combination of these four amino acid elongations / insertions may be used in the same beta strand or alpha helix or in different combinations of two or more beta strands and / or two or more alpha helices. The four amino acid elongations / insertions may be used in the one or more beta strands and / or one or more alpha helices consecutively or separated by existing sequences.

[0494] The one or more beta strands and / or one or more alpha helices may be elongated by one or more of TVTVTV, VTVTVT, VDVYTV, YTYNEK, TKTYNV, YTVTVK, ILTLNV, STLTVK, SVTVTV, LTLTVK, VSTIRV, VEITVT, SVTITV, ITITVK, VLNVRL, SEVTVR, TVTVTL, VTYTRR, VVEVKV, VEVKQT, TKTVKV, VVVTQR, RDIHTV, HTRDVV, TTIIDV, HKSTVK, VDTDTV, YTSTRT, TNTDTV, WTKTTK, VVTITV, STITVK, TTTITV, ITITRK, ISSNTI, NSNTVV, TTTNTI, NTVSVK, KYNYKV, YDYELT, TEKYEV, YIYEQR, VLTVRV, VTVTVK, VHTQNV, STQQQT, TQTQQV, QTQTTR, VDTNTV, QTSTTK, SKTNTV, QTTTIK, VYEVRI, LTARRT, STEVRI, VTLTQK, EVNYQV, SGENLT, YTETVK, ITNYKA, YTNTVV, KTTYTA, YTVSRR, VLSLKL, SDANTT, TRTLNL, LSLTVK, IDTVSV, VEKKTV, TVTVKV, VTKKTK, VTKSSV, STNTIQ, TTTSTV, SVLTQK, VYTYTV, VTYTQS, TTTYTV, YTYTSK, VRTNTV, TENNTV, TKTTNV, NTVTEK, VGTYTV, GTYTQT, KNTYTV, YTYTTK, VENSLV, YEKTRT, TEKITV, KTKTRK, ELTLSV, STLTTR, TTTLTV, LTLTVR, TVTTTI, NTTTLT, IIIIII, TTVSVK, TLTYDV, YNYTQN, NRTVTV, YTYSRK, VVNVSV, LNVNTT, TKTVNV, VTVSVK, TLNVRV, VTYNQK, STTVNV, VQWSVK, VVTVRL, AEVRVQ, TTTVRL, VTVLQR, TVNVNL, SQVLTT, TVQVLL, VNVTVK, IYTLSV, STITIK, SVSVNV, LSLSVK, KVNINL, SSIEEV, SKRIEL, ITVEVK, TVTTTL, STVVIT, TVTTVL, TTVTRK, IVQHTV, HTHTTQ, TRTHTV, HQHTER, VLSVIV, ISVTVT, VIVRVR, KNTVTV, ITENRV, SQTVNV, VTTSVR, STINLK, TTTINV, ITITLR, VNVKVV, TVTVEV, VKVTRR, VVTVYV, YRHNRV, RNTVNV, VTVTVR, VNTYTV, HVYNEK, TQTYNV, YTYTRK, TKRVTV, VNVTRT, TKTVTV, VTYTSR, RLTISV, STITVV, SVKITV, TLTITV, LTINVV, TKTINV, ITISTK, VLNYTV, VNVNVT, SVTYNV, VVTVTV, LTIRVA, INQIQV, SRTEQT, TNRIEV, IQYTSR, VVNVNV, TVTVSV, VVVTVR, IDVKTV, STSTQN, TTTKTV, KTQTVK, VTTLTV, YTDTDI, LTTTVK, IDIITV, STLART, SDTIAV, ITRSVK, VTYNDT, SKTINI, ITVSEK, IDETTI, STSKTT, TKKTDI, VSNTVR, VTTYSL, FTNQQV, TNTYQL, YTNTIK, ITENKV, STNSVR, TTSNSV, NKNTRR, IVNVTV, VNVTVT, SFTVTV, VTLSVK, VVTYTV, TTYTTT, SKTYTI, YTYSEK, IRKDTV, SSHTTV, SRTDTV, QVNSTK, IVEVQV, VRVEIR, SVEVEV, VKVAVK, VNTETR, TVSTQQ, TRTETR, STETQK, VDNVNV, SNHTRT, SDTVTI, STRSDK, VNTVTV, VNTNNT, SKTVNV, VTTSTK, VVTGTV, STGTLV, TVTGTV, GTGTVK, LTYTVS, TKTITV, ITTTVK, TVTITL, VTVTLT, SLTLTL, ITVSVK, TTNLDL, SSLKVV, SNTLKL, LTISVK, ISTYKL, YESKVT, DVEVKL, YTLKVK, VRKERK, SEEVVQ, SRTEVK, ETESRK, IVTVTV, VTIDQR, SVTVDV, ITVTVK, VTKITV, STTNTT, TRTINV, VTVKLR, EVTVKV, VTVTER, VTTVQA, VTSTVT, TTTVTA, VTNTVR, MVNISV, SNITIT, SVEITV,ITIKTR, VIEVTV, SKITIT, ISTYQV, YTYTTT, TRTVTV, YTYTTR, TVTITV, STLNVQ, TVTINV, VLNVRV, SSYTLT, VTYTTK, TVTISV, STIVTN, TKTIVI, ITITTR, VGTYSV, STTKQV, TETYKV, YTVETK, VYKVTV, VTVNLT, TVKVNV, VTINRK, VVTLRL, STVTVV, TVTLTL, LTVIVK, STVTVT, SKTVTV, VTITVK, TQVSTD, STKTER, TKTETD, KTKTEK, TLLVNV, SNLNVT, SVTVNV, LILSVK, ITTVTV, STITRT, TNTVTV, ITVKVR, VTNVTV, SNVTVT, TTTVTV, IVTVNV, VTVTTT, VTVTSK, VVNLTA, LNYTQK, SQTLTA, LTYSVK, TINVNV, VNVTVN, VNVEVK, VRLYTV, STYTEK, TRTYTV, YTLTVK, KVEKSR, SNENQV, TKTTNR, KKETVK, ILNVEA, LKGKRI, SKEVKA, VSFYTR, VETHTV, STSTRK, HTKQER, VNTLTV, VTKTLK, ARTLTV, LALSVK, KQTYTV, STATTT, TNTYTV, YTATTK, VYTFTV, VTTTLT, SSTFMV, IIISTR, STVTIS, GVTVTV, VTISVR, VREYKV, YTSTQT, TDDYTV, YKYTVR, EVTVTV, VTVNTK, TVTVNV, VNENTV, KTNTVT, TEKNTV, NTVTTK, TLNVTL, VTYNLV, TNTLNL, VTYTVR, VYTLDV, VTSTYI, QRTLTV, LTYTVR, INNATV, SSVTVK, TRTATV, ATVTVK, VNTETV, SKATDV, TNTETV, ETVQVK, LTVSTK, AKTVSV, VRTYTA, AEYVRK, TTTLVA, YTYSER, INNITV, YSNNTT, TKKINV, ITSEVK, INDRTV, SSSKTV, TNTRKV, SESTTK, VVTYKA, ATLTVT, TVSYTA, VLNTNV, SNDRRT, TDSTRV, TNVTVR, TVNITV, STINTT, TVSINV, IQITVR, IRTYTV, VTYTLT, KKTYTV, YTLTEK, TVVVTV, VNVQVV, EVKVQV, LTVAVK, VRQYSV, FTYTYT, YTYTVR, QVSVTV, VTVSRS, VTITER, LTATIT, ITVTVS, VTTVTL, STLSVN, SVTVTL, VTTVKV, VTITTT, KKTVTV, VTISVK, STVTET, TKKVTL, ILNVNV, SNINAR, SVDVNV, VNVSTK, VSTYAV, FTYTQT, SNTYTV, YTYSQK, IKNINV, ISNNIN, KKTINV, ITNTRS, VLNLNV, LNYTRT, SDTLNV, LTYTVK, VVTIDV, SRVTVV, QVTITV, ITVSVR, VTVTTA, LTLETI, TVTTEA, LTLRRR, VLTYTV, YTYTVT, TKLITV, YAHQVK, STETVV, NRTYTV, YTVSVK, VATVKI, VTSTKT, TTTVTI, VTYTIS, VQVWTV, STLVLV, TVTWVV, WTLTTK, VRNYTA, YNWNVK, NQTYNA, YTLSVK, IRTVTV, VTSVTT, SQKVVV, VTVQVR, INNYTV, STRVET, SNTYVV, YSRSTK, KDTVTI, VTNTTI, TKTVTI, VTNTVK, ITSNKK, HDHNDI, TRSDNK, HAVTVK, VTTNTV, NTNTTT, NETRTV, NTNTVK, VQEIRV, LEYKQT, TKEIKV, YTYRQK, VLNVSI, LSYNTK, AKTVNI, VTYSQK, STNTQT, RQTVTV, NTNTIK, VPTVTA, TDTVTG, YTNTQK, SNTVTG, YTNQTK, VTTVNA, VTTTVT, TVTVTA, VITTVR, STNTTI, ITITTK, VNTSIV, STSTVK, TNTSTI, SISTVK, VLTYTL, YSNNRT, TNTYNL, YTNTTR, VDNYMV, YTNNQV, STTSNA, YTSTRR, VVTVKV, IEHSRT, TDTVSV, VVTVTI, TTVVVV, TITVVI, LTSTVV, VLEVIV, SSVKIT, SVTVKV, ITYTQK, EEKNIV, NENTTI, TKENTV, NINRTR, VVSVSL, ANINVK, QVSVNL, VSITVR, QLVVKV, SQVETR, SVTVEV, VKVTVR, VNINTR, LTNNVQ, TTTNNR, NTNTRK, VVNLTV, SSLTIT, TSTLTV, LNLTSR, VTATTL, SSVTTK, ERTTTL, VTTTVK, ELTVKV, TTTVSV, VTMTNR, ERKYTV, YTYVEN, TKTYVV, YTLKER, LTNNTT, RTTVNV, LTNTVK, KAKYTV, VNVEVT, KRKYEV, YSLEVR, VGTITL, SQITIT, TVKITL, ISITVK, IDTNNV, NTNNVT, TVTNNV, NTNTVR, KDTTTV, SSNTRT, TQKTNV, TTSEVK, VDTITV, STKTTT, SETITV, DTRSVK, INTYTV, STITTK, TVTYTV, YTITVK, LTSVVK, SVTVVV, VISSVK, TVNVTV, VIVSTK, VKNNTT, SKNTNT, TKNNTT, NANTVK, VRTYNL, VTVTRK, TTTSTL, VVRVTV, STAVLT, TTTVVV, VEVTVK, ILTVNL, VTSNVK, TLTVNL, VTLTVK, VSNITV, IESNVI, KRTINV, LTLQVI, QKTVQA, VTLTVR, VNLKLT, VTYTAK, IATVQV, CRRDIV, SEDVVT, TRTDVV, DIVTIR, KVTVRV, VETNLK, SKKVNV, VTVEVK, QINITV, VSVNTR, TLSINV, VVTTTV, STVTVK, TVTTTV, TTVTVK, IYTGTL, GTGKLI, STSAKL, GTGSQR, VVTITI, TNTITI, ITISVK, LTVVVN, TVTVVL, VGTIDV, STISRR, SKKISV, ITIEVK,EITYTV, STYNNI, TTTYNV, INNYNV, YNYQKT, SRTYQV, YQYSIK, VDTSTV, TTVTRT, STSSVK, YEYKQT, TKENKV, YTVKEK, IVNVRV, VSVTVK, TVKVTV, VRNINV, VNSNDT, RKNINV, INDTTR, VDTYQT, NTRTRT, TDTYTT, YTRTVK, VLSYTV, STYTLR, YIYTTK, VKEYTV, STDKEI, TEEYKV, YTDRNK, VENIAV, VTTTTV, RKKITV, IAYQSR, VSTKTV, STETTN, TETKTV, KTETVK, IRTYKV, IEVTVR, TKTYTV, YTVTEK, IVSVSV, TLTVNV, VNVYKV, YTQETK, TKTHEV, YSQTQK, VVNVEV, SNVTRT, KTNVTV, VNVTVQ, IVTVRV, ATVTRT, SVTVTI, VTVSEK, VRTDKK, DTKTST, TKRDTK, DSVTER, VDNVTL, SNYTQT, SNTVTL, YTQSVK, STITVI, TVKITV, ITIEVR, VNTNNV, STNTVT, TRTNTV, NTNTTR, VDTHNV, HTNNRT, NKTHNV, HTSSLR, VDNYNV, VSNNQV, TRTYNV, YNHTDR, KETKTD, STKTVT, TVKITD, KTIEVR, VTIVQD, SEVKIT, TTEVKD, VIVKVK, VFEIEV, STIKTT, RKEIKV, IEISIK, ITEVTV, ISIKVT, SVEVKV, VRTYTV, YTYTQT, AETYTV, YTYKTR, VNTYTL, YTHNEK, TKTDNL, YTKTEK, VTVETK, TSIFTV, ITFVRE, KNTFDV, FVFTEK, VLNLSD, STQLKT, TRTELD, RNQTER, THTRTV, STRTEV, TRTDTV, RTVTTK, VVKVTA, LEVEVK, TTKVEA, VVVSVK, VNTHTV, HTHTTT, TNEHTV, HTERVK, TVQVQL, TTVKQT, STSVNL, VQVSEK, ILTITV, LTITVT, VLVLSV, LTWTVT, SRTLTV, LEVSVK, IDTSTV, STSTKI, SQTSTV, NTIVQV, STVNQT, TTTVNV, VIVQQR, VNTYRV, SETNTV, RRTYNV, YTTTVR, VLTLTL, AELTVR, SVTLTL, LTLSVR, VVNVSL, SNVNVV, TVSVNL, VQTATK, STQTQT, TVNATK, YTDQVK, RNTHTV, HTNTIV, TQTHTV, HTHSLK, STSTAV, TNTTTV, TTITVR, IYTVTL, VTVEKR, TTRVEL, VTVELR, VEEEIV, NQSTVK, TKSETV, ELRSQK, VATKTV, HTYTQR, SETKTV, KTYTVK, VDESTL, STSTKT, TETSTL, STKTVK, VYELKL, VTLNVT, SVTLNL, LKVSTR, VHNFTV, FSNTTT, RNNFTV, FNFDTR, VTKMTV, VTEEEI, SKTMEV, VKETTK, VSAQSQ, HNDTRT, SKRNDQ, KTDTSR, VLNVTV, SNVTTR, SVNVTV, VVTLRV, STLTIA, TSTLLV, LTVKVK, ITVRKL, IEINRI, ETTINL, ITVRVK, QRTVTV, VTNNLV, VTVTEK, VVNISV, VNAKYT, TTNIKV, INYTRK, TLTYRV, STLTLT, KTTISV, VTNSKT, TQTISV, ITNTVK, VATFSV, STYTLT, TQTVTV, FTLTVR, VVTTTL, STFNVV, TETTNL, VTTVTA, VTQQQT, TNTVQA, VTQTQK, VKNYTL, ASNNEI, TKKYNL, VTVEEK, ILTLTV, STVTTT, TLTKTV, LTVTTR, VVRTTV, SVTTTV, VTTQTV, SSQTRV, SDTQSV, QTQSVK, VYTLTV, STLTIT, TVTLTV, LTFTVK, VTKTSV, STNNRT, TGTTNV, NLNQTK, VVTVRV, VTVQVA, SVTVQV, TTVAVK, ILNINL, SEIKVV, SVTIKL, TETRRV, SEHTRV, TETRTV, HTRTVR, TRTVTL, VTVTLV, KRTVTV, ISVFTV, STGIVI, TVAFIV, FLYIVR, VINVTV, SRNVSV, VEVSVK, VKINTV, NTNNRS, KQTNNV, NLLSSR, IERRAE, SEDVTT, DNRRVE, RSVQVK, VTNIKV, SETKTT, TQEIKV, ILTTTK, VYNVKA, FTYKET, TRTLKA, FQYTVR, VRTYSV, FTYTET, YTYTER, ITEVSV, SELRVT, TVEVRV, VGVTVS, ISVSKV, SESKTV, TKTSKV, SIVTQK, VTTYNV, FTSNRT, SEKYNV, YSIVRR, VKVNTL, NSNRES, TKENRL, NTVTVK, VVRVSV, STVEQK, STTVEV, IVNITV, ITVTVR, TVTVQV, LTLTVV, TTVTIK, VDTYSV, SELKET, TKEQKV, YLRREK, VYNVTV, TTVNVT, TVNVNV, VTVKVK, TVVITI, VTSKLV, SSTIKI, IVSYDV, SSYTVT, VHTITV, STITVT, ITITVA, VESSSV, VSNKTM, SKSSKV, SSVEVK, IVNVNV, VTVKLT, TATVKV, VTVTDK, VTVTVN, TTTVTL, VNRYTL, ATYNEK, SKTYNL, YTKHER, IAEYKV, FKYNVS, SVEYYV, YLYKVK, IREYNL, GTYKRS, TSEYNL, YTYMVR, STGTVT, TNQVTV, VENYTV, YAYTQV, ITTLSV, TTSTTS, SKTLTV, LTSSTK, IEMYKV, QEYTKK, TETYTV, YTYRRK, VDTVTA, VSVTVV, RKTVSA, VTVTTR, QVRLTV, STLNVT, TNELNV, STNTTR, SVLSTV, STRQVR, VVEITV, STVKVT, SVEIKV, ISVSIR, HAKVRV, VEVGVT, TVGVGV, VDVTTK, IYTVKV,VVVKVV, VTVTTK, TTSFTV, STEVTV, VSVSVK, KVVISV, STITTV, SRTITV, ITISQK, VWTHTA, HTHTVT, TVTHTA, HTVSVK, IDKNTV, SNIETT, TTKNEV, TTQTVK, VSSYKV, YNNNIS, TNTYNV, YTNRVK, YTNTET, SDTYNV, YTNATK, TLNQRV, SESNLT, TQTQSV, QLSKVK, YTHTRT, SETYTV, YTQTTR, KYSYNV, GNINVT, SITYNV, YSLSVK, VTQYSA, STTTVT, SKQYTA, YTLRVR, VLTVSV, ATVNTT, IYTFTV, STFTLT, SKTFTV, FTVSIK, IVTTTV, SSSTTT, TKTTTV, TVTTVK, VDTYTV, DRSKRV, TDTYVV, LTAKVT, SKTIKV, ITLSER, VLEIDV, STLVIK, EVTIVV, LKLTVK, VEVEVI, TNKINV, VIVQIK, LTKVTV, SATTVK, VTTYSV, SSSTRV, LTLSVK, VATYTV, STYKIV, TRTYKV, VLLIKV, LEGVVV, SVKLVV, ILAQVR, LTFERR, TRSYEV, FTLTER, VKNENV, STKNVK, TDTENV, ELITVK, IQKNTN, STSEEK, EKKNEN, SSNKVK, LTTTMT, IIIIIV, IIIIIR, STTYTV, YTYSTK, VRNDDV, SEDVET, SKQDVV, DSDAVR, VVTVSV, SVLLTV, VTISVT, VGNLKT, SQLNVV, SVSLNT, VGSATA, LTGTVT, TVTATA, ATGTTR, VTTVTV, VTYRRV, TDTVRV, VTYSTK, VVSIRV, SEIIVK, SVTVIV, ITISIK, VKIYTV, KVTYTV, YTVTVR, VTYNRT, VTYSVK, VVKIIV, SEVTYV, VVTLTV, TRTLTV, LTITVK, TVTVTT, TTVQTV, TKTVQT, VYTYTL, YTYTVA, YTITER, LTTTNR, FTTTVR, VRTVTV, FNVTQT, VGSYRV, SEYTSV, SKTYTV, YTYSRR, KVTVTL, VTVEVV, GVTVEL, VTVSAK, VFTYAV, VLNLTL, TVKLTL, LTVTQR, IDTKSV, KKETTI, RRTKTV, KTFDQR, TLTIRV, STLNIT, ITATVK, VLTISV, STITLK, VEGKKV, ESKTTS, KTTRKK, STYNRT, NVTYNV, YTYSVK, VKTLKV, STNTET, TKTLTV, LTNTTR, FTVTLI, SVSVTV, VTVSVS, TTVVTL, ATVEVK, SKTVEL, VKVSVR, VLTITV, ITYTVK, IFSVEL, AKATIR, ASATVR, RNTIEV, STIDVN, TTKIDV, IENENK, VGIYRV, SEVKLT, NLEYKV, YIYAEK, STYTTT, ITESVK, QITSTV, STSAVI, TNTSAV, STVMVR, ATFTVK, TKTYIV, YAFTER, KGTYKV, SEYTLN, YTYSIK, STGTTI, RKTYTV, YTYSQR, ILTLDV, ATLTAR, LTLTQR, TQTYTV, YTYTQN, YIYSSK, VTTLTL, LTLNRE, TTTVNL, LTLKVK, VSTVTL, STVTVN, VERYRA, YEYERT, NTLYEA, YRYQTK, IVRVEV, VKVETI, RKSVEV, LKVTVR, VINYRV, YNYERT, TNNYNV, YNVTVK, FSYTVT, TKKITV, YTLEVR, IDTFTV, SITITV, VTRSVR, VETKTV, KTKTVR, SETTTV, KTVSVK, VLENKQ, STNKVT, TQENKQ, NKITTR, GTVTVK, SSTVTV, VQTYTL, TNTYTL, VAEVTV, STTVTV, VTVSVA, VEKRED, STREVT, SDKRED, RERKVR, VRTETV, STKTQT, TDTETV, ETTTVK, RVTYKV, STYEYR, KLAYEV, YKYELQ, IVKVSV, VNTTLT, EKTVTV, VTYTEK, KLNLTL, STSNVV, TTTLNL, LTITVR, IRTTKV, SEDTEV, YTDTVR, QVNITV, SETKRV, STEIKV, VTVSTK, VVNTTV, YTLVVK, SVRTVV, YEVDVR, VYTVTV, ITVTLT, TYTVTV, VVNVKV, IDVTVK, TVSVTV, VLEVTL, VSINVR, VTITQR, VVTIKA, LEVNVT, TVTINA, ITVTHK, VSFTQT, ITVSTK, VSTINV, SNDTVT, TNTITV, KVDTVK, ALTISV, SNLTVT, SVVITV, ITLKTR, KVTVTV, VTVSIK, VTEITA, LTNTQK, TNTITA, ITNTQR, VFTVKL, STVTVS, VNTQNT, NTQTYK, TKTQTT, QTETVT, VENYTA, FSYTRI, SEKYTA, YTVERK, SEVKTT, ERTVKV, VTVKTK, ATVTVT, INTVND, TTNTVT, TNTVTD, VTNQIK, VTKTII, VTVKVT, VVNINV, SKAIQK, TQNIIV, INVTVK, KISYTV, FNSVLS, SSTYVV, LTQSQK, KVTAEL, ATAVVV, TKTVVL, VVNATV, VSLTVK, TVTATV, QVQVSL, SSLTVT, TQQVTL, IQVSTR, EVTLTV, STLTIK, LTISSR, TVNVTQ, SEKTVV, TVRVTQ, EVVVTL, TEVKVI, TVEVKL, VDTVTV, STHTVE, YTHTER, STVTIR, YTVITK, NTLYSL, YSYNQT, TDEVNL, YTLRVK, TLQIYV, SAKTLN, VLISVR, VTRYQH, SEYSLT, TKTYSH, YIVTQK, IVRVRV, LDTNVT, TLVVNV, LRYLVR, VTNRTN, ESETRI, TQTRTN, ETETDR, INTITV, VTHTNS, YTMKSK, IYNFNA, LNSTLR, TRSFTA, FQYVSR, VETNTS, STTTTK, TRTNTS, SVTLNV, LTVSVR, VRNYRV,VNKTEV, YTKTER, VDTQTV, SEQTRV, TTTQTV, QTRTVR, VVKIKL, SEGTRQ, TETITL, ITVTRK, VYVHTD, HTHEKT, TNKHED, HVHTEK, ITNINI, FEHKTT, TQVIKI, INVEVK, INTKSV, STSNTT, KTTKNV, VTVVSV, STSTRV, NETVTV, ITTVSV, LTTNLT, KVTVNV, VTQTVR, VTNKTV, KNTTEI, TKTRTV, KTTTVK, VDNVSV, SNKTTT, RDTVTV, KNKTTR, VQTNTV, TTNTNA, SITTTV, NTNNEK, ITNLTV, SNITQT, STTLTV, LTISEK, TTLVKV, VESNVT, ITTSVK, ISVTVK, VNTNTV, VTNNRV, SETNNV, NTNSVK, TLTITE, STITTE, TKTITE, ITLTQR, VLTVTL, VRRITV, VTINTT, ITITIR, TVTTKV, NTSTVT, TRTTTV, NTVTRR, STVNVV, TKTTNL, TTVNVR, ATTTTV, SNNTVV, IETHSV, HTVTEK, EVTITA, LTTTTI, TRTITA, IKTTVK, KLIIIL, LKYTRI, TLTITL, IIYTEK, VDHVRV, TEVNRT, KTTVNV, VRVEVR, VVIVQI, STVQIT, TVEVQI, VIVKER, VDNINN, SNQTTT, TKTITN, QTQTQK, STRQQV, SETVQL, VTQSQR, VKVYTY, YKYELR, TRTEEI, VNTTTV, STSTQT, TQTTTV, STTAVR, FTTTRT, STTVTL, VTVNVV, TVKVNL, VVTIAV, ATVTIK, ILKLSV, LSLTLT, TVKLTV, LTLTTK, VSKRRK, SEEETK, TKTREK, RKETTK, VVNVND, SRVIVT, SVTVID, VAVATV, STTTVI, AVFTVR, STHTLL, VVRVRV, VTSETV, SRKVEV, VEVQTR, VVTVSG, VTVTIT, TVTVTG, ITYTEK, TNTFTV, VNTLSL, STYNVK, YTYTQS, VVNVRV, VDVKVT, TVSVKV, VSVVER, ERTETE, SKSTRV, TETETE, ETVTVK, IVQITV, VSVTVT, STSTVV, STTTVK, KTTARL, SELAVT, EVTAAL, ATVLVR, IITYDV, SSYNEQ, TRAYNV, YTYTQR, VEDYND, YNYNRT, TEDYTA, YKLTVK, KNVSTV, SQTSKV, SVSTVK, SEVTVT, VETITV, STLTRT, TTTIVV, VTVTNT, YTQQTR, ILVINV, VRVNLI, VNNVNV, TANNTK, NREVNV, SKSSRK, VDSKQV, TKTKTV, KTKTIK, VVNITN, SELKVT, SLTIKI, IILSSK, IDTQTV, SAQTRS, KQTQTV, QTRTIK, IKTYEV, YRYREI, TKEYRV, YEYLVK, INTTAV, ATYNQV, TKKTNV, TTIEQK, VTIVSV, SILTKT, VTTTVR, VDRYTL, TKTHTL, YTWTEK, IYSYEV, FRSNQR, SNTYNV, YRVFER, TTTVTD, SVTVTD, IDNNTI, STQTTK, STTNTI, QTTSTK, LTVTTK, TITILV, APVAVT, TVTIAV, ILVTTK, STVETR, TNTVEV, VVTVTL, GTLTTT, VFNVTV, STVKTT, VTVTVH, VYRVNV, VTLEVT, TKKVEV, VNVTVR, VSTVRV, VEITRR, TETVTV, VKNVKV, VSVRER, VIEINV, VNITVT, IKITVK, RVTISS, STNSVT, TRTISS, TITVTV, SAVSTT, SVTVSV, VITVTV, VVTLQL, SVKLLL, LEISEK, IIKIEV, STNNTK, TKEINV, ITIKES, STINVT, ITISVR, VGNVTV, VTDTIV, SQTVTV, VIDNVK, VDLQLS, SENKVV, TNTQKS, QLNTRR, IVQVTV, VTVLVR, TVQVLV, VTVIER, VTVTQK, VVNITV, VTVNTT, VTVEGD, TGEVTV, FVFVFT, TLVFVF, VEVKVT, TLEVEV, WWW, TVTVVV, ITITVT, TVKITI, ITITIT, TVEITI, TLEVVV, RTRTRE, KKTETR, ATATVT, TATATA, DTDTDI, TRRDRD, DTDTDT, TDTDTD, ITITVE, TAKITI, ITITFK, VTVTVV, ATATIT, TLTATA, LTLTVT, TLTLTL, FVFVVV, KVEFVF, TVTITI, LSLSVL, TISISL, ATATAT, TTTRQV, TNTQTQ, VIVIVI, TVSVIV, LTLTLT, LTLTYT, AVAVAV, TVQLVA, SVSVSV, FVFVLV, RLEFVF, VTVTIV, EVETEV, KKTKVK, TTTTVT, TVTKTL, IKIEIV, TIKIKI, LRLELT, TNNTGQ, TLRLRL, TITYTY, ITITIV, TIKLTI, KTKTKV, TETETK, LTLTIV, IITITL, NTNTTE, TTKNTN, TLTVTV, ATATVV, LTLTLV, TATLTL, VTVTVL, QTQTQT, RDTDTQ, TTVTVT, YTYTYT, TLTLTY, VVTLTL, VTLTIV, TYTITI, TLVVVV, DTDTIT, TVTDTD, TVKVEV, ETETRT, TNKETE, LTLTLE, TATITI, DVDVDV, TKVNVD, TLTITI, KVQDTK, TLRVTV, LVLVIT, TVTLVL, IQITII, TVVIQI, LELKVV, KIELEL, RVTVTV, TATVTV, FTFTVT, TVTFTF, ITITVI, DTDTVV, TKTATD, IVIVIV, TVEIVI, VVVVVT, TVEVVV, RVTITI, KVTLTL, IQITIT, NVQVQI, TSKIVI, ITNTNT, TKTITI, HTHTHV, TKTLTH, IVIKLT, KLEIVI, IKIKIT, KVEIKI, KIKKKV, KDEKIK, ITITVV, VKVKVE,TLKVKV, VSVPVP, ITITID, TVRIRI, STSTVT, TTTNTN, FTFTLT, TATFTF, VTVTVI, IVIVVV, TMVIVI, RTRTRT, TEKRTR, ETETVV, TRTKTK, HTHTHT, TATYTH, KVEITI, LVLTAV, IQIQIT, TVQIQI, VEVEVV, TVKVKV, ITITLT, KLTITI, YTYTYV, VISYTY, TVELTL, LILSLT, TVSLIL, TVTYTY, QTQSTT, TTSQTQ, TKTKTD, GTVTVT, TFTGTG, LILILT, TIKLIL, ITVTVT, TTTHTH, TLTLVL, TITITV, TTTTLV, TGLTTT, VTVTMV, TGTVTV, LTLTLI, LVTLTL, ITIEIT, TASATA, TLKATA, RVEVTV, TAVLTL, KGTITI, KVTITI, ITITIN, VVKLTL, NVTVTI, LTLTIT, IEIKVI, KIEIEI, TSVIVI, SIKTKT, ATATLT, LTLTLQ, RTRTET, TKTRTR, EVEVVV, TKVEVE, DTDTDV, TTTDTD, EEETEV, IREEEE, RLTTTT, GTGTVT, TLTITG, TVTATG, TTTLTL, ITVTVV, YTLTLT, TTTYTY, KITLTL, TLVLVV, TVKVVV, TVVVVV, TVTYTL, DTDTVT, TTRDTD, ITFTFT, RLEVTV, TKTNTD, TAVTTV, IIIIII, TVTATT, NTNTNV, TSTNTN, KVKVKV, TKSKVK, AKAEIT, VVKAKA, TITLTL, TAKATA, RTRTRV, TETRTR, DIDILI, TRVRVD, VVETET, VIVIVE, TIKVIV, TVEVKV, TVTNTN, VQVQVV, KATLTL, LALSIV, TVSVAL, TVTLTA, AVAVVV, TVTAVA, TAIIII, TKTQTD, V

[0495]

[0496] EVKVV, VSVSVV, TLTVSV, ITISIT, VKVEVE, KLKLKV, LVLVLV, TGVLVL, EEETRT, TEEEEE, TITITI, KLTVTV, VTVTSV, TLTYTY, TKTKTK, TLMLTL, VTLTLT, TVQVTV, VDVDVK, TKDVDV, ATATAV, TLSATA, SVSQST, TKQEVE, NTNTKT, TNTNTN, TLQLTL, NTNTNT, TKTNTN, ETETEV, MTMTVV, TVTMTM, TLVITI, HEHRTR, VKEKEK, TNTTTT, KTKTKT, TEQKTK, HTHTKV, TQTTTT, LVLVIV, AVATAT, TSTAVA, DEDEID, TTNTEN, TDEDED, NTNTVV, TRTNTN, TVVIVI, TATLTV, VTVTRV, TDTRTV, LELRVV, TLELEL, IVIVAT, VITIVI, HTHTVV, TRTHTH, AVATLV, TATAVA, YEYKIT, KSEIEY, TIKVTI, VVVTVT, TVQVVV, TTTIVI, FTLTIT, LKKETE, TATVTI, TIKVTV, TKTLTL, KTKTIT, LKTNTK, VRVEKT, EERVRV, TVTWTA, YVYVYT, TATYVY, KVELTL, VQVQVT, TVKVQV, VVVVVR, STSTSR, TVTSTS, TIEIEI, LTLTFT, TGTLTL, ITITIK, LTLSLC, TLSVTL, VTVQTV, VTVTVD, QGQVTV, VIVTVV, LELELE, RLKLKL, TKTNTH, TLQYTP, QRTRTR, TVVAVA, HIHTLV, TVTHTH, GTGTLT, TLTGTG, LELTLT, TLTAEL, TLTLVI, NTNTAV, TVTVTN, EVETKT, KQTVVE, RLELEL, TVRVTI, TLTLIL, LELTVI, TVKLEL, YTYTIT, ETETEI, QIQEQE, TVTVTI, KITITI, TLTVTL, LVRVVT, TEVRVR, IRIEVD, GLRIRI, TVSFVI, VTVTAT, TVVLVI, ETETET, TQTRTR, TLQLQL, QVQTTV, TVTQTQ, AATLTL, YSYSIV, TASYSY, VTVTIE, AVAVIV, IVIVVT, YTYTAT, TLKVTY, AIATAT, TITATA, KKKKKT, SGGVGA, TKEKKK, HIHIHT, TTIHIH, NTNTVT, KATVTV, VSVSVT, TCSVSV, VKVTVT, LTVTVT, TVELTV, IVTITI, TTATAT, IQIQIV, KVEATA, IVITIT, QVTIVI, KIKIKI, TKIKIK, KYEITI, TTNTER, TVLITI, AEAKVV, TVTAEA, TVLVTV, NKNTNT, KTKTKI, TETKTK, VKVELT, TMVVVV, VEVEVL, TLKVEV, TLTAVA, TVEVEV, KTKTQV, TDTDTK, ATATVR, TVEVTA, ITITIE, TLTLTI, LTLTAV, ITISIV, TVKIKI, VKVEVV, KLKTKV, TGTATA, AIAIAK, TVSAIA, TTTSTS, LVLVVV, TLELVL, IEITTT, TVKIEI, TLVIVV, VEVTVV, TETKAV, TLKTET, TGKVTV, TTTTKV, KVTTTT, LILILI, KIEIIL, VEVKIV, TTTKTK, AQAQAT, TLQLQA, FTFTIT, EQEQVT, TKQKQK, LILIVV, TIILIL, VTVEVT, AKAEAT, TIKIKA, IEIEII, LVLVVR, TVELVL, NVNVNV, TTTRVN, ETETRR, TLLETE, VVTVTA, VEVTVR, TVLVEV, TAQLTL, DLDQRV, ETRDLD, FKFKIE, TVKIKF, KVTVTA, VTVSVV, VEVTII, KVEVEV, NVNVTV, TKVNVN, TAMLTL, LVLVLT, KLTLVL, LILTIT, TQTQVN, SYEVQA, AEAEVI, VVEAEA, LKLELD, EMKLKL, TKTDTD, KTTVTV, LTATAT, QKQTQE, TKKKKQ, TGKLTL, IEITIT, EVKIEI, MSMSML, TISASM, YQYTYN, QSQYQY, TTTTVV, TVTLTT, KVKVVI, IKTETE, VLTITI, STSQSV, TTTETS, TIRITI,VKVTMT, EIKVKV, YTYTLT, TKEYTY, RMRTIL, TRMRMR, QTQTTV, THTQTQ, VKVEIE, TGKVKV, GTGTVV, TLTLTV, VIVIII, TIIVIV, VQVTVT, TLAVQV, TAEVVV, QTQTTT, TKTETQ, KIKIKT, KKEKIK, TATLTI, KLTLTV, IEIEIE, VVKIKI, VEVKVI, KVTVTI, YTYTID, TKKYTY, IQIQVL, YTYTYE, TLKYTY, WTWTVV, TATWTW, LILELT, NTLTLT, AIAIVI, TIIAIA, TVVLVL, IEIEVR, VGYTSN, TVSIEI, TLTATI, VQVQIT, TVQVQV, DRDEKT, RDERRD, TITVVV, TTTITI, LQLQLV, TAQLQL, TTTTET, TKTTTT, AIATVV, TKTATA, KKEKTY, RVRTRV, TETRVR, IKIEIT, TVTIKI, TLKITI, DTDTET, TGTRTR, TVSIVI, HVHTQT, KKTQVQ, TVTVLV, TLKIKI, TLKVTV, HQHTIT, EKTHQH, TNTNNG, VTVTLQ, TLSVTV, TATTTT, IEIKIV, RVEIEI, ATLTLV, TTTATA, LTLSLT, ATATAR, ILELTA, TVQVNV, TVKIVI, ITITFT, ATATIV, TQTITI, NVTLTL, STSTTT, TKTHTS, YTYTIV, KTKTHT, TDTKTK, ITITVN, AEAKAV, TIEAEA, TVQSTS, TVQLTL, TKTQTN, YTYTYQ, VKTYTY, KVKVKT, TKQKVK, RKTKTK, TIKLTL, VVVVFV, TNQITI, TSKVTV, TETDTD, TSTLTA, DITLTL, LVKLTL, VVVTVE, TNINVN, TVEVEL, ITLTVT, RTRTYN, RTRTDT, AVATVV, RREDTD, VTVTVE, FTFTVV, RVEFTF, EERETE, VVVTLT, TATVVV, TLQVQV, KIKITI, KALRTR, ITITLV, TDTVTV, KEEETK, VTVQVV, ATATLV, TATVTA, TVSLTL, IVIVLV, VSTIVI, AVATVI, LTITVV, ITITAI, TVTITT, TIKITI, KVKTTV, TKTDVD, TLELVA, SNGTGL, TVTETI, SVSVVT, TVTVVT, HVHTHV, TTVHVH, IQITVT, TVTVQI, AMAMIV, TVMAMA, LVLVII, KIKVKV, AKAELT, TVKLKL, KRELTY, LQLQLT, TLTLQL, VTATAT, VGTVTV, VVVVVI, TLVLVL, TLQLTA, VTLTIT, KTKTTV, KKEKTK, VDVKVK, TITVDV, KIEVEV, ITITIQ, KKELTL, QTQTRV, TDTQTQ, TVRLTL, AVATLL, TVYAYA, TLQVTV, TVTAVS, VIVIIK, EVEVIV, AIAIAV, NVIAIA, EKTKTK, LILTLT, KTTNTN, VVVSVV, KVEVVV, TAIVIV, VTVTLL, LTLTAT, RVTVTI, TVKVKI, TITTTT, TTTTIV, TITITT, ATATYT, LILILV, TAILIL, YIYKVK, VKEYEY, TVTTTT, NININI, TKININ, KQKQKL, TKQDQK, TKTFTI, VLTVTV, TNNTGT, IIIIIV, TVTVII, RSEATI, LQLQVT, TVQLQL, KTTDTD, ALTLTL, LTLTVI, TVTVTR, TVTITA, TVILIA, LTLELI, ELRLRL, VTVTRT, TLNVNV, AKAKVT, TVKLKA, TVTLQL, GVGVLV, TAELEL, RIDIDV, TRIRIR, ITIKIT, TETYTY, RIDLTL, TVQLTI, TLVIVI, AVITIT, ATAEAD, TVRARA, ITITAV, KTEITI, ITITHT, TLKLTL, KTKTKK, TILKTK, DTDTTV, FIFSIV, KVVYVY, LTLTTT, TVTATL, VIVTLT, AIKIIV, NVNVVV, VTVNIN, SDSQTC, IVAVAV, IVIEVE, TIRLRI, KTKTLV, TFIKTK, TVKNTN, TATATY, AVAIAI, TAKYVA, VNTTTL, ITIQIV, TAQITI, TVSISI, DTDTRT, TQTTTD, LELEQT, TTELEL, ATATAI, RVEATA, TATFVF, ITITMT, VTVTST, LLTLTL, VVVVVK, ELEVVV, SNETQL, VTVKVE, TKTYTY, IKITLV, TLTIKI, HEHKVV, TREHEH, ATATAE, LVLTLV, TLRLVL, TEKKEK, TKTETE, TATITL, KLTLTL, VTVTIK, LTLSLV, AEAKVT, TAEAEA, TVIVTV, TISVTV, TRVYTY, TVELTF, QIKLEL, YQYQYT, TYQYQY, VTLTVT, NTNTYT, VTVTKV, STTSVK, LKLEIV, TVTLKL, STSTVA, GTATIT, TITGTG, SISISV, TAISIS, KVKIKI, VVVKVT, TAEITI, VLQLQL, INITIK, TITIEI, EVKVKV, LVGGKP, ANANGV, GGPSGG, SRSSGG, LVNGVP, ADASGN, SPATGG, LVNGKP, AGASGK, GGGYQS, TVDGLP, SAASGF, ATPEEG, GPGGGG, LVDGRP, GGGPFT, RVNGEP, VVGGKP, ANEKGI, GGPNGG, LVNGEP, IVGGKP, HDGLGG, GGAGGG, TVDGRV, VVDGTL, ADATLR, GVSAGG, STLKVQ, VNSGTA, GSGTGK, SGSGGG, SVNGVR, VNNNTT, VNNGTV, TNNTTT, SGSFNG, NTNNTK, IVNRVN, SDELIR, SVKRGN, SVDEVQ, VINGTA, GNGTGK, SGSGSG, LTNDVR, VTTGTA, TTTGGG, SIIIII, VTNGVA, SNTTNL, SDETLS, SGTQGG, SVESVK, VNNNVV, NNDLLK, TGGNGV,NVRGES, INNNNN, SNNTGS, GNTNNG, NTDNVA, VTSGTN, SNNELK, TGTTNN, SGTTVN, VVNGNV, ADATGK, TGTGTG, ATAGVQ, VTENKV, VNVKLT, GGEVGA, VKVGEK, VSSGVV, SNDNLT, SGSGNG, SVDGVK, VVNGTV, ANVTVK, TVTGTG, ATVTVQ, VTSGTA, SNNTLV, SGSVNG, STDGVK, VTNGTA, ATGNGV, GGTSNG, ATGQVQ, TVNGTT, GNGNGK, GGTGNG, GSGGVK, LSGTGK, TGTGGG, IVNNTA, TSSTLK, and SVSVSA, VTVGVQ, GTGVQA, YQSRVK, VNQYQV and YTYKQT. The one or more beta strands and / or one or more alpha helices may comprise one or more of the listed insertions. Different combination of these six amino acid elongations / insertions may be used in the same beta strand or alpha helix or in different combinations of two or more beta strands and / or two or more alpha helices. The six amino acid elongations / insertions may be used in the one or more beta strands and / or one or more alpha helices consecutively or separated by existing sequences.

[0497] The one or more beta strands and / or one or more alpha helices may be elongated by one or more of TVTVTVTV, VTVTVTVT, VRIHVYTD, STSEFTER, TQTHEYND, FIYVETVK, INTSSYTV, SSSTYTLK, SETSTYTV, STYSYSSK, VVTSTYSV, HTSTITQT, SRTSTYTV, HTYTVTEK, VTTSKVKA, SESEVEVT, STKSEVVA, NTSKVSVK, RTTSTYAV, STSKVDVQ, TVTTKYTV, STYTVTVR, VSTVSINI, SAVQINVT, SKTVQINI, VTVSNSVK, IRTITYNV, VSSEITER, SKSVKIEV, ITYTETIK, VATTTVNH, STQTVTTT, SKTWTVNH, VTVTLTVR, VVKFNVEA, VSVEVTVK, SVKVEVEA, FKVNVEVK, VNINTFRV, SENSFTLS, TEKNSFNV, NIFTFTER, TLISTYTV, SKSKVNVK, TKESKVNV, YIYTLSIR, VATATVNV, VSSTVTTI, SYTATVNV, VTATTQYK, TVTSSVRV, SDSTVTVS, SRTSTVNV, VTVSVTER, VDTSTKTV, STSTRTVT, SETITVNI, STKTVSTK, ELNININT, SNNLINVK, KTTILINT, ININYTVK, VSSYEYRI, STYKYKLT, SKSYKYNI, YKYEFSSK, MQTSTVTL, SSSDVTVT, TVKSSVNL, STMTVIVK, VVTVTVNL, STVTVTVN, SVTVTVNL, VTVTISRK, KTTVTVTD, SGLTVTVT, SKTVTVND, VTVTVSVR, IESVLATV, LTLQAKTT, NREVQATV, VSALSRTR, ITTVTITI, STSTVEQK, TKTVTVNI, VTITQTQK, IYTSTVNV, ISSQVTET, TTTSQVTV, ITVTVQEK, TLTVTVTV, STVTVVVD, TTTVTVTV, VTVTVTEK, VVEVTYTI, STVKVTVK, STTVKYNI, VEVTVSQK, VVTVTLAA, LNLTVTTT, TVTTTLNA, VTLTVTVR, VINVNVSV, VGVEVKLT, TSSLEVNV, VNVNVTLR, ISSSEYNL, SNSKLLTR, TVSSKYNL, SSYELTER, KVTVTVSA, STVTVVVN, SVTITVNA, VTVTVSVK, VETSNITI, SNSSLTRT, TEEYSITI, STINQRVK, VLSITVYY, VKSLVTLK, TRTILVNY, VSITVSEK, IRSIFYSV, SSISYTQK, SLTISYNV, ISYLDSVK, TRTVNKTL, SNSSKTET, HRSVSKEL, VTVNEDVK, VVEKKITI, STNVVTET, SKTKVINI, NEIKVTTR, VNTNTIKV, STNTKTIT, TNTNTIEV, NTITSTVK, SIKTEVDN, SKKEVIVN, TVKTEVDN, VKVEVRVK, VLSIKISV, SSIEITYT, SVKIEINV, VSIKYLVK, VDTNTLNL, STSTLTTK, TRTNTLNL, ITITKTVK, TNTVTVSI, STSTVTVQ, TVTRTVNI, VNTSTYQH, STSTVTET, RKTSTYTH, STYTVSEK, VYTSTYTV, STSTVTTR, SKTSTYTV, VTYTVTVR, TDVSLITI, STSQKQTV, TESSQINI, SLHLQTER, VGTVTISI, VNVTVNVV, TVTVTITI, VSITVTEQ, VVTSVVKH, SESQVEKR, TVKTQVTH, STVVVESR, IEEDTKTL, STEKKTRK, SETEKKNL, DEKTKSVK, TTTLTYRL, SELTVTRT, TTTVTYNL, LTLTVTQK, VLTIRASL, SQSVATTT, SKEIVANL, ITIRTLTR, VDANNNTH, SNSNNTVT, SKKNNNNH, SNNNNEQK, VRTLSYRI, VGYTYTET, TTNKTYNI, VTYSLSEK,VVTSTVTV, STSEVTVV, TVKSEVNV, STVTVTVK, VLVVVVNV, VTVEVRQQ, TRTVEVNV, VVVVLQQK, VDISEYTV, NSSRRTLT, TRTDRYNT, NIYERTVK, VLTVSQNV, STITQTQT, SRKDTQKV, VTISQDVK, VNTSIYTV, STSTYTET, SRKSTYNV, STYIKSQK, VATTTYSV, STSKYTLR, SVETKYTV, FTYTLLQK, VATYTYEV, SKSTYTET, TRTYTYQV, YTYTLQER, VTVYTYRD, STYQHQIT, TTTYQYSD, YHYTHTVK, KVSVTVTV, STVTVTVT, SETVTVNV, VSVTYSQR, VNIYTYTV, YTYTYKQT, SKTYTYNV, YIYTYEQK, VLTSTVTV, STSTVKQQ, TQTITVTV, STVTVEQK, VDASTYRV, NNESITRV, TTTTSYNV, NAYTRIVK, KTTNTYTL, NTNTREET, STTNTYTL, NTNTESLK, VRTVTYKV, SEVVYTET, STTVVYNV, VTYTESVK, TNTSTVRV, STSTVTTV, TNTSTVTV, SIIIIIЕР, VDMINVSV, SSVTVRLI, STTITVSV, IMVNRSIK, VATVTYKV, SEVEVTRR, SVTAEYMV, VTYTVSER, VLTITVTV, STVTVTVS, SVTITVNV, VTVTYRSK, VNIHQTTY, SSHTTKKE, STTHTTNY, HITQKKEK, VGTQTDTR, STRTVTTT, TRTETDNR, QTVTNKVK, NDVTSINI, VSNDNNLR, TNKNDINI, VLKSNEVR, INTLTNTL, STLLSKTV, NKETLNNL, LTNTSNVR, VDTFTYTL, STFRYTTT, TKTFRYNL, FTYTNTTK, QLTVTVTV, VTVTVTVV, VTVTVTVK, TDNSTYSY, SSSVNTTT, TNTSVYNY, STYTRSIK, VDTNTNNV, NNNTNTTT, SKTNTNNV, NTNTTTTK, VSIVSVTL, ANVSVTTK, TRTVSVNL, VIVSTTEK, TKSYTYRV, SEYQYTQT, TKTYQYNV, YSYTYTSK, EVTVKVHV, ISSTVTRK, TDTVTVLV, ITVKVTVK, VDVTTNTN, STNTNTLT, NKTQTNNN, NVNTNSVK, VFTSTVNI, STVINQKQ, KETSIVNI, STVTKEQK, VIKETKTN, STEEKTRT, TEKEEKTN, EKKTIQRK, RESTRTNT, SNSDQERV, SEVTDQTT, SSTRVEVK, VDTSKNTV, STSSNTTK, TKTSSNNV, STNKRTSK, VDTVTVRV, LNVTVVNS, SKTVTINV, LTVTVKVA, VTTLTLTL, STLTLTRR, STTVTLTL, LTLTRSRR, VYSYTYNA, ITSTITVK, TVTYTINA, YSYTVTVK, VYVVRYTV, STSEYERV, NNTVEYNV, VVYRYSVK, VVTSTISL, VNVQITQQ, TTTSQITL, VTVTITQK, VSTSINRA, SEATQTRV, TSTTTNRA, STNILTVK, VLTTTLTL, STSTLTTE, KKTTTLNL, LTLTLLER, VNEQLVSV, SNQEVKET, SRSDEVSV, QEVLNSVK, KYSYNYNV, SNSEHTES, KDKYEYNV, YSYNNETR, VNMSTHRI, STSTHKTV, TVTLTHNI, SMHEKTLR, VKSYSERR, YEYEEKEV, TRSYEENR, YSESETVK, VITDTVNV, SESSVTTT, TRTDSVNV, STVTVTTR, VDVSTYSV, SVSTNTQV, RNKSTYNV, SVYTTEVR, KDSLLYNV, SSSNYTTT, TNSHNYNV, ITYLSTTR, IQESNRSE, STSRRITT, SKASRRNE, STSNTATK, TEAYRYSV, SESEYTRT, TETVEYNV, YTYRYTRR, VLKLTLKI, STSSLTRR, SETKSLTI, LKLTRAEK, TLTSTIYV, SSSEITEK, TNTVEINV, STITLTVK, VRSFTYRV, STYSINLK, SRTFSYNV, FSYTIAEK, VVTVTVNV, STVTVTVV, SVTLTVNV, VTVTVSMK, VSTLTYNV, SNLTYTLT, SRTLTYNV, LTYTYSEK, VDNHTYTV, HTHSNTTT, STTNSYNV, HTHTSAVK, INIHTQSN, STSEVKER, TIEREQSN, EIYTNRTR, VKTYEYTV, STSKVTQV, TKTYKYNV, YNYEVTEK, IDTKTKSE, STSTDTTQ, KRTKTDTE, SSKTTSVR, VDKFTYRF, SESEYLEN, KRTIEYNF, FKYTYTER, VVTTSYRD, STHTYTVT, GVKTTYND, ITYSYESK, VVTVTLNL, STLTITTT, SNTVTINL, YTLTSSTR, IDIIEVNV, SSVKVEVK, SETIKVNV, VLVEHSVK, VATYEYSL, YTYGYVDR, ESKYGYNL, YTYEVEMK, VNTFKYQH, SNHEYQET, SKKFEYNH, FTYKDETR, VDISRLNV, DESRLVTR, TRESRLNV, DIYRSRTR, VQVSTYRV, STSTYELK, TQTSTYTV, SVYTETRK, VVAVTLAL, STSELTLI, TVRLELNL, VALTLRVR, VLEVKVSV, SEVTTVVK, SVIVTTSV, VEVKLELK, KVVITINV, SSSAITTN, TKTIAINV, VTITITEK, TATVTVRV, VEVLVTKT,TTTVLVNV, VQNYSYTL, SNYTVTQT, NTNYTYNL, YNYSLSTK, KVTVEYKL, STVKVEVT, NVTVKYTL, VTVEVSVK, VNTSEYTV, FTSTYQQE, SKKSTYTV, FTYEQSEK, VLISKVSV, SDSEVKLT, SQESEVNV, SIVKLSTK, IARSEVNL, STSTVTVT, SRTSTVNL, YRVEVAEK, VTTFTYSL, SNFTYTVT, TRENTYIL, FTYTVTEK, VATSKYTV, STSEYEVK, TRTTEYTV, STYKYTEK, VVVVKVTR, SESEVTLK, SVLVEVTR, VVVKVEQK, VNKNTVNI, NSNSVTVS, TVTKSVNI, NKVTDTIK, VWTVKVSI, SNSTVTVT, RVEVTVEI, VTVKYQSK, IFTNEYSV, TTATMTVT, RRKLTYMV, VTYEAEVR, NQTISVTV, SSITITNN, TKTVTINV, ITVSNTVK, VETVKYSL, FSYENTKR, TERQEYNL, VTYKELVR, VVTITINI, STSTIKVV, ERTVTINI, ITITIKVK, VKSSTESL, STSTETTV, SRSNTENL, SSETESVK, VESYTYSV, STSVITKT, SDNYVINV, YSYTLYTK, VETYTYNL, SNYENKNI, TKTYEYNL, YTYTNTIR, RQLTMKKK, DTKVNDES, KQTNVQTK, TLKMNSVK, VVTVTVTL, VTSTVTVT, SETLTVNL, VTVTVSCK, QTSIKITL, SESSIKVT, TVEISITL, ISIKVKVK, VDQSSHRV, STTTHRRT, TDSSTHNV, SQHSRTVK, RISVTYNV, STVSYTDT, SRTISYNV, VTYTYASK, VNTYTYTV, YTYTYTTT, DTTYTLTV, YTYTTSVK, TISITVNV, STVSVTVT, TVTISVNV, ISVTVRMR, VSTSTYTL, YTYTYKLR, TEESTYNL, YTYTYKRK, NQNYTWSL, SSSTLWRT, TTTYTLTL, VNWTETVK, VSTSLTSS, STSETNNS, SNSSETNS, TTSLNSER, IYEKKKKT, SEKEFKVN, SKEKEKVT, KEKKFKEK, VLEHRVTL, HTHSNVVV, SITNSSTL, HEVRVSVK, IEASEYNV, STSKYVVT, TKEIKYNV, SAYEHTTK, VIVVTISA, VTITVTVT, TKKVTISA, VVITVLEK, LNMSRYSA, SSSELTTR, TDESEYNA, SMYRQKRR, VDITTTSH, SSTTTTVQ, STSVTTTH, TTTTNSRR, VTTTTHTA, LAHTVVVT, TVTTTVTA, LTHTVTVR, VQSTTYTL, STSTNNST, RNDTTYNL, NSYKNSNR, ILTIVISL, STIVIRVT, SVTVVINL, ITIVISVK, IEKYEYSL, STYIEEKK, SETYEYNL, YKYVESRK, QVTLDYSV, SNLVVVLT, TVDLVYNV, LTYDVRTK, VVTVRVSV, SEVTVTVT, TVEVTVNV, VTVRVTVR, VYTTVYNV, STSTYTYR, TRTTTYTV, NTYVYTVR, VNKVTINV, PKSEITVT, SVKVEINV, VKITIERR, RNTIEYRK, STIRYEMV, TKRERYTK, ISYEMEQR, VNTETKTT, NTTTLTKR, QETETKTT, NTKTLTVR, VVEVKTNV, VSTVNVEK, KKTVVTSV, VETKHEVR, TTSVSYAI, TNITVTVN, TKAVTYNI, VSYSVTVK, KDTTTVTV, STSEVTTL, TRKTEVNV, TTVTVEQK, VYTFEFSV, STFTYERT, TTKVTFNV, FTFEYQLK, VNERTENV, SSISETEK, TKTRSENV, YEQTESVK, VERSEYSV, STSVNTRK, TETDVYTV, SRYENTRK, IQESKYRV, SESTQKQI, TNESTYTV, SEYKQKEK, VNTYEYRQ, SNYKYEEK, TKKRKYTQ, YTYEETTK, VVTVTYKL, STVTITVK, TVTRTYNL, VTYTVTVK, VDSSKKNN, SESEKTVR, SKKSIKNN, STKKKQER, YTYTAEQQ, RTTYTYTV, YTYTQTQK, VLTVTVTV, TTVTVTLT, TKRLTVNV, VTVTHIEK, VNQSVNQL, SQSQNLQK, SKTSQNDL, SQTVSQEK, LETSTYRT, SESTVEMR, TKRETYNT, STYTVQHR, VRENKTTV, SSNKEKRS, SLEKKTTV, NENKEAEK, ISTDTRTH, DTDKVEVV, TKKIKRTH, DTDTELKK, VDTTTVTL, STITVTRT, RVTTTVTL, VTVTVARR, VETITYTV, SNYTYTRT, TETITYTV, ITYTYTQK, IVMSTTQI, STSTVNLK, TSTATTVI, SMTTVTKK, VDTSTVSV, STSDVTVE, RTSSDVTV, STVTVTRR, IVRVEVTV, VTVTVEQR, TTKVTVTV, VTVERSVS, VVTVTVNA, VTVVVTIK, SKTVVVNA, VTVTYSSK, IANYTYTV, YTYEYTTV, TQAYEYNV, YTYTLLVK, VDTSTYTL, SNSTYTRT, SVTSTYTL, STYTRNER, VNVNTNSV, NTNTNNVK, TTTNTNNV, NVNTNTVK, VVTVSIKD, STIEITVK, SVTVEIED, VTISVSVK, VTVSTVTL, SSVTVTVT, SKTSTVTL, VVVTVQSK, VLVSTYRV, SESSYKET,TRTLSYTV, YVYTYTQK, VNTSINKH, STSGNTIV, TQTTGNNH, DTSINTVR, VNTSTYTD, STSTNNVQ, STTSTNND, STYTNSVK, VKTLAKKN, SQSVKTTT, TKEVVKTN, ETQADKTV, VVTVTVTV, SRTATVNV, VTVTVTQR, VRTYTYSV, STYTYTET, RVTYTYNV, YTYTESVK, IEKIEYTI, STSEVEVI, TKKIEYVI, IKYELEIR, VVTYNYHL, ATVTITVS, TVNKTYTL, VTYNISVR, VFTVIVSV, STVQVTVT, RTTVQTNV, VTVIVTIR, VERSEYTV, SNSEYELV, TERSEYNV, SRYEVRER, QEINENSQ, NRSRRQLT, TRRERNEQ, NIEERLEK, TLLIEISV, SKIKIVRK, TVEIKINV, ILIEARVR, VVTVSVNV, SNVTVTVQ, SVTVTVNV, VTVSVSVR, VTTITVTI, SNVTVTTK, TVSVTVNI, ITVTTTTK, VVNITINA, FNISITVT, TVTVSINA, INITISIK, VTTVTVSI, SNVEVTTR, STTVEVNI, VTVTVSLK, VKTSQESL, FESTETVT, TKKSTETL, FTEQLEEK, VYTSSYKA, SESTYTRI, TTSITYTA, STSSYTIK, VKIIRYTV, FESSYTMI, TNEISYNV, FIYRSRVK, VLNSTINI, STSTITNT, SRTITINI, SNITISQK, VVSTNYNL, SNSTVVTT, SVSVTYSL, TTYNVQTK, RIRSTTTV, STSETLEE, KKRSETNV, SRSTHVER, VLTTEVRV, STSTVTLK, ILLILVNV, VTVELTVK, IYTSVYKD, SESTYTIT, TKEVTYND, STYVIIQK, VQTSTANA, VTVTATTT, SETKTANA, VTATQHTT, VVTVNVNL, SNVTVKVT, SVNVTVNL, VTVNVDVK, VYVSKLRL, SESTLTVV, TVTSTLNL, VVLKRTVK, TTVSTHNL, SAHMVTTT, TTTSMHTL, VTHTVTTR, VETSTYTV, STYKYTRT, TETEKYTV, STYTVTTK, VVKSKTPI, SESTTITK, TNTSTTNI, SKSKVTKK, VNQSTNTL, STSNNKNV, NRTNNNTL, SQITNSEK, VNNLVYTA, LTVTYAKT, TNTLTYTA, LNIVQTTK, VATVTISV, STSTITTT, AKTVTINV, ITITYYAR, VGTFTYNA, LTSTFTLT, SKKFTYLA, FTYTLRVK, KKEVNYTI, SNVKYSTT, TKEVKYNI, VEYNIRQK, VESSISTT, HTHQHQRE, TETSQSTT, HSVIRRSK, VVTVTIKV, STVTIKVT, NVTVTINV, VTITISTK, VETVIYSV, SNSTYVRT, SETVTYNV, VTYIATVK, IVTVGVSL, SPVIVTVT, SVTVIVNL, VTVGVTVK, VQVTKTKV, SESKSIVR, STTTKTTV, SVTKNSVK, VDTKTYHV, AEKTLNVN, TVTKTYNV, KTYTHTVR, VVKSTLKD, STSKLTTK, TKTEKLTD, SELTITTK, QNTNIDTR, SSNVDVVT, TKTNVDSR, NTNIVTEK, VNTTTVTE, STSTVTTT, SNTTTVTE, LTVTTSTK, ITSIKVSV, IKVEVTTK, SREIEVDV, ISVKTKVK, VNTNTNTN, STNENTVN, EVRTEITN, NTNTNLVK, IRSSSINV, SNSKRTLT, SETSKINV, SSISESTK, VALSTYQH, SSSTYTVT, TVTSTYSH, SLYTLTVK, VLKIENSV, SNIINERR, ENTVINNV, IKTERRVR, VDKSSYNI, SQSEYTIV, GKKTEYNI, YKYSISEK, VITVVVQV, VTVVVSQK, KKTTTASV, STITQQQQ, TQTTTANV, VTVTQSTK, VKTSTLTL, SSVEVKVI, SVESELTL, VTVTVLVK, VVKVTVNL, SSTTVTQQ, TKTVTVNL, VKVTVTQK, RVEVSVDV, VNVSVVET, TRSVSVSV, VLVSVTER, VLEIKIDL, SSSEVKVQ, TVTVEVNL, VEIKVTVK, VVTISISV, LTSSIIVK, QVTISINV, ITISISVK, IKTSTVTL, SSVTVTTR, TRTSTVSL, KEVFEYKV, TDKFTYNV, FVYEYQVK, IFEKKYTV, STNIIKLT, TTEKIYNV, NEYKIRLR, VNKYSYNL, SNVSYKVT, TEEYSYNL, YKYSTTSR, VVTTTTSA, LTSTTVRR, SVTTTTSA, TTTTNRRK, TLKIEINV, VSVSITRT, TVEISINV, VKIELTRK, VATTTVTL, STTTVTTT, TTTKTVNL, TLVTYTSR, VDIFTYRN, STFTYKRT, SRTVTYNL, FIYTQSEK, VVTVTYSH, SNSQYKVE, TNTVQYTH, VTYTVTEK, VVTVKVTH, SNVTVTVT, KVPVTVTH, VTVKVEVK, VLLLTYNL, STTTYQYT, TKTLTYNL, LLYTYQQR, VNQSTSTV, STSSQGVK, SKNSSQNV, QQSTGSTR, LLTSTITA, VTITITTK, TEESTITA, ITITVRTQ, RREIVRDL, SRIERTED, TVREERNL, IEIVDTVR, IEVIKEKV, SNSEEKRT, RDEREEDV, IVEKRITR, VVSSTVTI, SSSTVVVT, TVGSTVNI, VSTTVITK, ITTSEYRQ, SKNIQIQK, TTKSIYEQ, STYEQEIR, TVTTTISV, SNSTITTK,STTATINV, TTITVSVK, VVTITVSV, STVTVTKT, TVRITVTV, VTITVTVK, VRVTNDDL, SNSTDTVK, TETTTDNL, DVDNDTKK, VQTSTYTV, STSTVTLV, TKTETYNV, STYTVQTK, VSSSTYQV, SNSTYTVQ, TQTSTYNV, SSYTYTQR, KNTSDNKL, SRSNNNTV, SQTSNNQL, SASDNSTK, VLEVNNTV, VSSNNKSV, TRSVNNNV, VENNLTEK, VVVSRYTF, STSTYTLT, SVTSTYNF, SVYRYTVR, ISTVDVSV, SDLEVVDQ, TRTVEVTV, VTVDVLEK, VLSVRVNV, LSSTLTIT, TVSVTLNV, VSVRLEVR, RVEITVTV, ITITLTLV, TTTLTVNV, IEVTISLK, SKVKVTER, SREVKVDV, VIVTYYSR, VVEIKIRV, SSSEITVK, STKIEISV, IEIKVEVK, VQTYTYSV, STYTLTTT, TRQYTLRV, YTYTTQTK, INTVNYQV, VSKEDTQV, SQKVEYTV, VTYNNQVR, VNSVTISV, STVDNREN, SRQVDNNV, VSVTNRNR, VDTITIST, SKSTITRV, TTTITINT, ITITKRVK, VTTSTVRL, SSSTVTKT, TTSSTVNL, KTSTVTEK, INVYTYTV, YTYEVTTT, TTKYEVNV, YVYTTTTR, VELLQYNV, STSTYRET, TRTVTYNV, VLYQRTEK, VVTVTARV, ITLTATTT, SNTVTANV, VTLTVQTK, VREYKYTV, YTYTIKQQ, TTQYTINV, YTYKELQR, VSSSTHIV, HSSTHTLK, STTSTHNV, HSHTHSEK, TVTVTVNV, INTITYSV, STSTQTTT, KNSITQTV, ETYTRTER, VRTSTVNV, STSVNNQT, TNTSVVNV, STVTNTTK, VQRYIYTD, STYEYEKI, TRRYEYND, YRYILEEK, VNVSTITE, STSSITVT, TKTSSINE, NVNTVTVK, VVTSVRND, STSTVTLT, TLQLTRND, STSVTIVK, VKINKHRS, STNLHEMR, TNKTLHTS, NIHKLEVK, VDTSIYKV, SDSTYVRK, SLTSTYNV, YTYVATQR, VTVIELKV, STSEYTET, SRKIELNV, IVLEYTVK, TVNVTLKV, SQLTVART, TTSVTLEV, VRLTVQTK, VFESKYTV, STSEYKVT, NESSEYTV, STYKYDRK, KRENIQRV, SESSQKEI, TRENSQRV, SEIIETEK, VVTVEYTA, VTVVVTVV, SVTVVYSA, VTVEVSVR, VVASEVNV, STSIVTVK, TVTSIVNV, NEVEVTVK, IDENKNDA, NTNKNKKT,

[0498] TNENKNNA, NENKRKEK, TDEIKYSV, SSISYKKT, SNTISYNV, IPYKRQVR, VVTVEVNL, STIIIKVK, ITVEITVK, VEEYTYRV, STYSYKRV, RETRSYNV, YEYTHARK, IVGVTTTV, KATVTTEV, VKVTVSVK, RIESSNTV, STSTNTTK, EKNTTNTV, SEVSVTTK, IKNVTVKV, SSVEVKVT, TKEVEVTV, VTVTVKTK, KMQYTYTV, STNTYNQN, SNTKTYNV, YTYTYSEK, KVTSTYTY, SSSTYNRT, TETLTYNY, ITNTNTTK, VNIITYNV, SKSEKRTT, SVKNEYNV, IIYTNSTK, KVKLEVNI, VSSVVETT, KRKVVVNI, LKVEITTQ, VSNLSYEV, SEYDYTRK, TKTLDYDV, YNYSYTVK, TDIYEYKV, YEYKYTRT, TQSLKYTV, YTYEYTSK, TVTSSVTV, SSSTVTVV, TVNSTVNV, STSSVTVR, KLQFKYSN, STSVYDVD, TKTFVYNN, FQYKYTEK, VKTISYNL, SSSTYTEQ, TKTITYNL, ITYSYTEK, VNTVHVNL, VTVTVKTT, SRDVTVNL, VTVHVSEK, VLTVTLNV, STLTLNQV, SQTVTLNV, VTLTQTQR, VVISTYNL, SDSEIEVT, TVKSEYNL, SIYTIQVK, VITATVSV, SNVTVTLT, TVTVTLNV, ATVTSTTR, INKVTVQV, VTSTVTNN, TSTYTVNV, VKVTNKEK, VDKVTYSL, STSEVERT, SDKVEYTL, LKYTQLRR, VNESNDRD, SESTKTTT, KQRSTDND, SESNKTTS, VDTITVSV, STVTITVT, SVTITINV, ITVTISRR, VTAQQYRH, SSSNQQQT, STKQTYNH, QTYQQQQK, VGTFVYTV, STYEVTVT, TVGFEYSV, FTYVVVTR, VVTVSYRV, SESVVKLT, TVDVVYNV, VTVSVRRR, IVSISITL, STISITVS, SVTISISL, ISISYSQK, KVTVSISV, VTVTIEVT, SVKVTINV, VTVSFLSK, VTTYEYYN, SNSKYTQV, TTKYKYQN, STYEDERK, IDKSVHRI, STSEITYK, SDENEIDI, SKHVHKER, VDKKTKTV, STSVTERT, TVKEVTTV, SKKTEIEK, IVVSTSTV, STSTVKVV, TVESTSNV, SVSTVKVK, VGTTQYNV, STYSYQRI, SETTSYNV, VTYQQSRK, TITSRISL, SDSEITLT, TLQSEINL, ITIRIKIR, VDTSRYSL, VNSTVERR,TTKNTYTL, VTYREERR, ENLSNVKA, LQVKVKNI, KNTSKVNA, VTVNNTQQ, VQENRKRV, SDNRKLNK, TTTNRKNV, NENRDTRK, VTSSNYRL, LESTYTTT, SKSSTYVL, YSYNTDVK,

[0499] VNTTTTTL, STSTTTET, TKTETTTL, TTYTSTEK, QRTVTVRV, VTVTVELR, SVKVTVNV, VTVTYEVR, IRTYTYNV, STYTYTQD, TRRVTYNV, YTYTYTER, TNEVKVRV, SEVTSEKT, KETTTSTV, VTVKRSVK, VVTITYQY, SNITITVV, SVTVTYNY, ITITITVK, TVVVTVSL, STVVVTVT, TVTTVVNL, VVVTVTLK, VLTSTLNV, ITSTLTTT, TQTSTLNV, LTLTTTTK, KITVSVTA, STVTVNHI, TEEVTVNA, VTVSYLDR, VHTVEYNA, LTVTITVT, TKRVTYNA, VTYEITER, VVTSTVTH, TVKSTVTH, VDTNSKRH, SQTNTKTH, STKSTSIK, TVTITISI, SSSSITVR, STRISINI, ITITIEVR, VLVNQYTD, STSQYTRR, TTTVQYAD, NVYQETVR, ILTSQVSV, VSVTVNLT, TTNSTVNV, VTVQNTTR, VLRINITV, STSEIEVV, STKVEITV, IRINLSVR, VNTNQYTN, NQNMNNQT, SQTNMNNN, NTNQNSQK, INISTNND, SKSVNNEK, TNTRVNND, SIITNTEK, VKEIRIEV, SRIEYKLI, SKEIEINV, IEIRYSKK, VLTITIDV, STITITKR, TVTITINV, ITITIKVR, VETSVKTL, SNWKKTTT, KETSKKTL, KTKVKSVK, VLSVNYSL, ADVHYENT, TRSVHYNL, VSYNFTEK, VNTVTIKV, SQSVITNK, SDTVVINV, ITVTSSVR, VINVIIKD, SEIIINRT, TVEVIIND, INIIYKRK, QLRANVNV, VSYDKLES, STTQDVTV, VTVLSQSR, VTSSQKTV, SSSTKQQQ, NETVTKNV, STKQQSQK, TLTVEYTV, STSKVTVT, TLTVKYEV, VTYEVTSS, VDITTVTI, VSTTVNNK, TKTTTVTI, VIKTNSTR, TLKVEVRV, SNVVLEVT, KTTLVVNV, VSVSLTVK, VAVSKYSF, YSYELNVV, TLTSEYNF, YVYKLTVK, IVNIIVNV, VTINVNTQ, SVTINVNV, INVIVKQR, VQTYSYSV, SSSRYTQK, TTNTQYNV, YAYSETVK, LLTWRYTI, SSVEFQQK, SLTWEYNI, LTYTFAQK, KLTTTVTL, SVTTTVTL, VSVTYSVK, IETNSNQN, SSNTNTLV, SRSNTNDN, NSNSTKEN, VKESEARL, VSSLDVET, SKEELANL, VEVKDQTR, VVTSTYTH, STSTNTTR, STTSTNNH, STYTNSRR, VEQSTISN, TRQSTVTN, SQSTVTVK, VTTSNVSL, SSSSVEQR, TTTSSVNL, STSNVTRK, VTTITITS, STIVNKQT, TKTIVINS, ITIKQTTK, VHTSTYSL, STSKNTVN, SKTDKNTL, STYTNSTK, VLESTIRL, SESTIRRV, SETSTISL, SEITVSVR, VRTSRVTI, NTSEDTET, TRTSEVNI, NIVRDTVR, VVSVTYTL, SQVVVTLR, SVTVVYTL, VSYTFEDR, VTSYTYNA, TLYTVTTT, STTYTYNA, YTYTVTVK, KRLYTYTL, SNYTYEET, SRKYTYNL, YLYTYEEK, VTTIEITV, SSVSIKVT, KVAISINV, ITIEIEVK, TTTVTGNV, STSTGTTN, SKTVTGNV, VTGTNSEK, VVEVRINH, SSILISTT, KKEVLINH, IEIRVAVK, IMESRYSV, STSMKKET, STEEMYSV, KEYRKKER, VVNVTVNV, ITSEVKTE, TKTVEVNV, LTVTVTTR, RRINMNTV, STNSEEVT, TKSNSNTV, MINMEVTK, VEENKYKV, SENEYKRT, TQLNEYKV, NEYKVLEK, IQTNTNSL, SSNTNKQN, TNEYTNNL, NTNTNKNK, VDEVEERV, SEVREERI, SKRVRENV, VEVERFER, VYTYNYDV, YSYTYTQT, TRTETYNV, YTYNYTTR, IVSVSVTL, VNVQVTLT, TVSVQVNL, VSVSVLVK, KASSETRR, SNSHVEVV, SVKSHVNR, SSTEVEVK, VITVTINV, STVTITVQ, STQVTINV, VTITITLK, VVTVTNTL, STSTNNNV, AKTVTNNL, VTITNSVR, VVTVAISV, TTVIITVT, SVKVIISV, VTVAVHEK, VRTIKVSL, ISITVTEK, TRTVTVTL, VNSVSYTR, SSVNVQTT, TNQVNYTR, VSYSVISK, TVTITIKV, FNITITQT, TTKITINI, ITVTITTK, VLTTTVTV, SSSTVTVK, SVTTTVNV, VKTSTVNV, SNSTVTVK, TRTTTVNV, STITYTEK, VLAVDYQI, SNLEVKVT, TTKVEYNI, VAYIVQRK, VVTVTVRA, VTVEVTVQ, SVAVEVTA, VTVTVTQK, INTSTNRA, STSTNTVV, SNTSTNTA, NTSTNSVR, VMTDTKKN, NESTKITT, SKTETKTN, NTNTTSVR, VTTVTHTA,SENELKQT, TTTQEHNA, VTHTLTQK, VLTVTVNV, STSMVTLT, RRNVMVNV, VTVTTSVK, RIVVRVKV, VEVEVEIV, TVRVEVNV, VVVRVEVR, VLNITISV, STSQIVES, TRKIQINV, VNITVEEK, VNTSSENV, SASRERVV, TRESRETV, STSSEEVK, VQVSTYSV, STSVVTEK, TRTSVYNV, SVYTETTK, VETSTLTL, SSSTLTRV, TTTSTLNL, STLTRTRR, VSTFKYNL, FTSEWKQE, TTTFEYNL, FNYKETVR, TNSSSVND, SSSTVTRK, SVSVTVND, STVSVTRK, TVKVTVSA, STSEATIT, TRTVELNA, VKVTVTVK, IVDVTVNV, SSIKVTKT, TTTNKVNV, IDVTITEK, IDTSTYNL, STDTYNTT, KKTSTYNL, STYTHTTK, VETKIFSL, SNSKDTKT, SNEKKDNL, TTKITKEK, VNTSNYSI, SSSNNTVR, STTSNYNI, ETYSVSRR, KSEVTVSV, SKVTFTLT, RTKVTVNV, VKVTFELK, IDTSTYTE, STSTNTRR, TDTSTNNE, STYTNQRR, LSVTVTTT, TVTVTVTL, VTVTTSSK, TVQSRYTV, YTSEYVTR, TKTSEYTV, YQYRYTDK, TVMTTVSV, KTTTTVAV, IMVTVTIT, VTILKLNL, SSLELKVS, SVELELNL, LILKYQEK, TVQLQLQL, TTLQYNTT, TKTLQAGL, LQLQYSTK, VTISTVNV, TVTTTVNV, SITTVTRS, VDTVTVSL, VNVTVTTT, TNTVTVNL, VTVTVTSK, VLTVALTL, VSLELKVQ, TVNVELNL, LTVAFTQR, KVSSSYSV, SSSAVEKI, NKKSAYNV, SSYSKDIR, RYRYEYKH, SESVYTET, SQKYVYTH, YRYEEETK, VVENKNNS, STNTNTVK, TVTNTNTS, NENKETSR, VNEITANV, STSTATLR, TQTITANV, VEATLLRK, VTTNTITV, STNANTTQ, TNTNAINV, NTNTNQVK, RVMVMVSL, SNVVVELT, TLTVVVNL, VMVMVTTK, TVTVTISV, STSTIEVT, TRKVTINV, ITITVEER, VVTVKVQV, VTVKVSVK, VVKSSVTV, SNSNVEVK, AVTTRVNV, FSVEVNTR, VTTVTVTV, ITSTITRV, SETVTIEV, VTVTIHET, VNNSEYQL, SESKYINK, TNTSKYNL, SNYENQTK, VVQVKVNV, LTVEVKTT, RKEVEVNV, LQVKVSTK, VNVSTSTL, SSSQSQNN, SETSQSNL, TISTSSNK, TSTIGIKV, LESELEIK, SKKVELTV, ITIGVSEK, RVVITVTL, STITVSVN, SVSITVTL, IVITVSVK, VETKTYQA, STKTETEK, TTTKTYDA, KTKTETVK, VEIVQVTL, LTVQVKRT, TLTVQVNL, LIVQKSVK, TAVVTYNI, SSVTVTVV, TITTTYNI, VVTTRYQT, STSEVVDT, SRVEEYNT, STYRVAIK, VVTSTVSV, STSTVTQR, TTTSTVNV, TTVTVSQR, RVQIDYTL, SSVRVTVV, HVEIRYSL, IQYMVSVK, VLQLTLTL, STSTLLVR, TTTLTLNL, LQLTLTVR, VFKVENRI, SLKENERV, TRRVENNI, VKIERTEK, VVTVTVSL, QTSEVTVT, SVSVEVTL, TTVTVSVK, TMTVKINL, SVITVTKV, SVTNTINL, VTIKVHVK, KTTTTITV, VNISITLT, KLTTSITV, VTITVEQK, VVTSSISV, VTVTITTR, SRSSTINV, VTISTSVK, VSTSNYNV, YNSTLTIN, TTTSTQTV, YTFNITVR, VDESSISV, STIKITRR, TLTSKITV, IEISRSVK, KETTEYTV, TTGKYTQI, TTDTKYTV, TTYEQTSK, VAVININI, SNIVNNKT, NENIVINI, IVINNTTK, ITTTTTNNN, STSTNTLT, STTTTNNN, STITIHVK, IRSVTVSL, AQLTVTTV, SQTVTVNL, VSVTISVR, VLTTNVQV, VNTTVTRK, TTVNKTRK, VFTSTNSL, SESVNTTK, TTTSVNTL, STNTSTER, VVRVSVNS, SRVMVVVT, TVEVMVDS, VRVSIKVS, TRSYTLTL, STYSLTEQ, RRTHSLDL, YSYTETQR, ILKVTVSV, VSTEVREV, VKVTVKVK, SINVSVSI, TRVNVSVN, TVSVNVTI, VNVSVTNK, TLTVTVQL, VTVTLSVK, VDSVTVKA, ISVTVVTQ, SNSVTVNA, VSVTVHQK, LQQHTYEV, HNHEITTT, TVQHEYTV, HQYTIQTR, VDTINYNV, SNIQITNN, TQTIQYNV, ITYNNNRK, VVISTITV, STSIINLT, TLTSIINV, SIITITLK, ILEVSLKN, SSATLKLT, SQEVTLNN, VEVSLKVK, VTTNTNRN, SNSTNTLS, KQTNTNNN, NTNTNTMK, VLKVEVSL, STSEVTYE, SVKVEVNL, VKVEYQER, VTTVTVNI, VTSTVTRA, TETVTVNI, VTVTVTVR, SKTVTVNV, VTVSVSRK, VDTKTESA, ATKTETKT, TETKTETA, VTKTRTTK,VESHSLKN, HESQLKKV, SDTHQLTN, HSLSKQQR, KDVYSYNV, YSYNYERV, RVKVEYNV, YVYRREEK, RTTTNYNV, STSKYDKV, SENTKYTV, YTYNYSEK, VKTLTYNV, SSSTYQVT, KESLTYTV, LTYNYEDK, VVVVTVTV, STSTVVVR, VVVTVSRR, VFTYSYRL, SEYTYTKQ, TETVTYNL, YTYTYTRK, VNTVRINI, SDTKITIK, TKTVKITI, TTVRITEK, VTTVKLTL, ATLEAKTT, TKTVKLNL, ATLETTER, IEINTYTV, STSIYTTR, TKTNIYNV, YIYTTTTR, VYTVNVNL, VNVTIEVR, TTTVTVNL, VTVNITVK, ITPITVRV, LSIEVTVT, TVQAEVNV, ITVTVTSK, TNVTKSTL, STFEEKEV, SRSTESTL, VVSKESVK, VETVTVSA, GKVTVTRN, TETTTVEA, VTVTESQK, VNESVYSV, STSKVKTT, TTESKYNV, SQHVTTTK, VDESKSRV, SDSVVKVV, TKTSVSNV, SESKVTQK, VDTFTYQV, SEFTYTKV, SDTFTYQV, FTYTRSVK, IVTVTVNL, STATVTVV, SVKVTVNL, VTVTISVK, VEKSKVKV, SESEVTRT, RNESEVNV, VKKKTTVK, VTVGVQTV, NIGTGVQA, MTMSITIT, RTEVSITM, KSKTKTKT, TDTKTKNK, ITISITIT, TITISITI, ITITITIT, DAKITITI, TVTISITI, VEVSVEVK, VNEVSVEV, LITITITI, ITISITIK, VITISITI, VTVSVTVT, TINVSVTV, ATATATAT, VATATATA, VTVSVTVK, DVTVSVSV, VVVSVTLT, ILTVSVTV, KVTVTVTV, TVEVSVEV, ENKSKTKE, KETESKTE, ITYSYTVT, TLSISYTY, TVTVSVTV, TVKVSVTV, IEITAEAT, KITITIRI, VITVSVTV, IVIEIVVK, ITEIEIVI, KLTISITI, IEINIEIT, NLNINILI, ISISISVT, TLKISISI, VTVKVEVE, NIKVKVEV, VTITITVT, VTVAVTAT, VVTVAVTV, AQYSYQYT, VNSISYQY, ITISITVT, TISISITI, VVVSVVVV, TLKVSVVV, VTFSFTFT, NATVSFDV, VTVAVTVK, RVEVAVTV, DTDSSTTE, ERKDSRTY, YSYSYSYT, VQQSSYQY, KITITITI, VTVSVTIV, VISVSVTV, YTYVYSEK, VTVKVTVE, RVTVKVTV, NTISITIK, INENSIIN, TITVSVTV, VEVRVEVV, ERKVRVEV, IVISLVLT, LLVISIVI, VTVKVTVT, TVKVKVTV, KEKSVEVT, ISKKSKEK, FVFSFVFT, TLTFSFVF, ATATITIT, DSKVTVTA, VLMVSVTV, RVTVSVTV, VTVAVTIT, TVTVAVTV, NTNKNTNK, EKTNKNTN, ITYSYTYT, KKDYSYKI, TITITITI, NLNSNLNL, TSSNSNLV, VVVSVVVT, VLQVSVVV, VLVSILIK, ELSVSVLV, VTVSATAT, TLVVSVTV, VTVSVQVT, DVTVSVTV, VKEKSITI, DVTVSITV, IVISNVNT, INSISIVI, DIDSYTYT, TVTDSITD, ATASATIT, TITISATA, VTASVSIT, KLTVSVSV, VTVSVTVV, VVTVSVTV, VTTITITI, QTQTQKQT, VKTQTQEQ, TTVSVTVT, VGRASVEV, KSKSKSVT, REEKSKSK, DKDEDKDT, VDEEEDKK, VKVSVKIT, VVTVSVEV, EKRSRTRT, VDTTVRTQ, ITISITIE, VTVTVTLT, TITVTVTV, YVYKYEVE, IIKYKYEY, VDVTLILV, VLTVTVIV, QVSVSKNV, DTDSRTRT, TDTDSRTD, ITITITIK, EITITITI, VERYVYSL, VEVKVEVE, VVKVKVEV, EKKESVTI, RTRTRTRT, RDTRTRTR, NQNSNQIK, VTENSNQN, VRVEVRVI, RKEVEVEV, VTVTVTIT, ITITKTKT, IDTITITI, VSVSVSVT, TVSVSVSV, GTGRGTVE, RAVGRGTG, TILITITI, VEVSVSVR, TAEVSVTV, EKKKTVTV, VTVTVSVT, ELTVTVTA, KVKSKKKI, VKEKSKEK, QTKTVTVT, KETKTKTK, NTNSVTVT, TKTNSNTN, YTYTITIT, TITYTYTY, VLVSVLIT, TVTVSVLV, FTFSFTFT, TATFSFTF, DTDADTDE, RETDADTD, NRNTNRIT, VNTSTNEN, YQYSYQYI, IRKYSYQY, NTNSKTKT, TSTNSNTN, VVVTVVVQ, NLVVTVVV, TATATVLV, RVEVSVLV, VITVTVTV, KKTVSVKV, EITISITI, IKVTVEVT, NVKVTVKV, VATISITI, IVLSLVLT, TLTISIVI, ATAAVTVR, RVEAAATA, IEISIEIT, LVKISIKI, VAVAVAVV, VAAVAVAV, ITISLTLK, IIEISITI, VKLKVTVD, VGRVKVEV, VTITITIT, NVTVTITS, VTVSITIT, QTQTKTKT, NKEQTYTQ, ATASATVT, RATASATA, HIHSHQIT, IEQHSHTH, ITVSVTVT, VLSVSVTV, YEYAYEYE, VEKEAYKI, VIVSVTVT, VLTVSVTV, MTMSMTVK, VVEMSMLM, KVTVTVSV, LITISITI, IEIKIEII, ILKIKIII, VTVSVTVQ,KKTVSVTV, LTLSLTLT, TLTLSLTL, VVVTVVVV, VLTVTVVV, ITITITDT, NSAITISI, VTVDVTVR, RVDVDVTV, NITITITS, YTYTYVIK, EYTYTYMY, TINISINI, VTVTITIT, KITVTVKV, QTQSQTQI, QNTQSQTQ, LTLTLTLT, TLTLTLTL, VTLSLTLS, NATVSLTV, VVVTVVVT, VVTVTVVV, NVNTIVIT, VTTNTNAN, ITISFTFT, YLTISITI, DSDSVSVV, LTSRSHSD, KTKTKTIT, VKKETETE, ETEAKTKT, TNIEAKTE, LTLTLTVT, TVTLTLTL, STKSKTKT, QTTSSKTS, FTFTFTIT, TTTFTFTF, NITVSVTV, ITISITIV, TITISIKI, STDTDTDV, EDTDTDTD, KTKAVTVT, LKVKAVTK, KVTLTVVV, YTVSVTVT, TLTYSVTY, VTVSVSVT, KVTKSVDV, VTVAVTVT, LVTVAVHV, LLLQLNLV, LVQVQLNL, VTVSLNLR, ELEVSLNV, ITIKFTFE, TAKIKITI, TLTVSVTV, IKITIKIT, IITITIEI, VVVSVVVE, VLKVSVVV, ILTLSLKL, RITITITK, IAISIAIT, DLTISIAI, VIVSVNVT, LLTVSVIV, ITISLTLE, TLRISITI, VAQVSVTV, DTDTITIT, TTTDTDTD, KLTKTVTV, YTYTYTVV, SRRIEYNL, TMTYTYTY, ENESENIT, NKVESKNE, ITITITIV, ITISIKIE, SVSTSVST, NSTSTSVS, EQTVSVVV, ITITITLT, TVTITITI, ATASLTLT, TLTASATA, ASASISIT, VISASVSA, LTVSVTVT, HTHAHTTT, RATHAHTH, FTFTFTFT, VVTFTFSF, VTIEITIT, KMEVEIKV, VVSVSVTV, HLHTHLVQ, VETHTHLH, VTVTVTVA, DDTRTITI, HTHTHTHT, TITNTHTD, NLTISITI, VTTATVVV, ITISITII, VIVTVIVT, TLTVTVIV, GVGSGVVV, KTTGSGVG, EGTVTVEV, ATASATAV, VAVASAEA, DRDEDRDR, VDEDEDRD, NVTVSVTV, EITITITA, QTQSQTQR, RQDQSQTQ, VLVSVQVT, TVKVSVQV, VTVQVTVT, KATVQVVV, NLTVTVTV, ISIKISIT, RIEIKIEI, VSVDVSVR, EVDVDVSV, ETTVTVEV, KTKVKTKT, DDEDVDTK, ITIAITIR, DIEIAITI, TLEISIRI, KVTVKVTV, GVGSIKIV, KVEGSGIG, ETETYTYT, TKTETETE, VTTITIVI, ITITITVT, KITITISI, VNVSVNKT, NVTVSVNV, VVVSVVVK, VVEVSVVV, AQASANAT, TLTASANA, NLTITIQI, VVVSYVYT, TLTVSVKV, TVTVTVVV, EVKTKTKV, IDTETKTK, VTVQVTVE, VLRVQVTV, VTVSVSVK, TTTVSVTV, VTLTLTLT, TVTVTLTC, TATVTVTV, VLVSVLVT, VISVSVLV, ATASITIT, TVEVSATA, VVTVKVEV, TTTTTTTIT, TVTKTKTK, TNTVSVTV, VTVAITIE, RLAVAVTV, LTLTFTVT, VATLTLTL, ITITLTLT, LVTITITI, NTNTNTNT, VVVEVVVK, LIEVEVVV, KKTKTNTN, YVYSYVYT, VEQYSYVY, ITVTVTVT, VVTVTVLI, HVHTHVVV, EKRHRHVH, VTVSLTLT, RVSVSVAV, ATARVTVE, VTRARATA, HTHSHTHT, TSEHSHTH, VVKVAVNV, YTYEYEVT, VITSSVTN, VKVSLKLK, EVEVSVKV, IVTVTVTV, VKVSVKVE, KVTVTVEV, VKVSVKVT, VVKVSVKV, VTVEVKVK, VVEVEVKV, KITISITI, VVVTLVLV, VLTVTRVV, VTTTTVTH, YTYTYTVT, DKTETYTY, ELESELET, LKEISELN, VSISISIE, RVTVSVSV, VTVRVEVE, HTRVRVEV, TTTTTTTT, TATTTTTTT, YTYDVTVR, DVDVDVTY, VEVTFKFT, TLTVTVNV, TFTVSVTV, YTYSRNRV, RRSESEVY, VVVSIVIV, VVSVSVVV, TLLVSVIV, ITIQVTVT, TLQIQITI, VIKISITI, DVDTDVTV, KDTRTRER, KVKTRVET, VKSKTKVK, DVVVSVVV, LKLQIKIK, ELLLQLEL, ITIKITIT, ELKIKITI, TKTKSVTV, VEVSVEVT, QVSVSVRV, VTLQVTVE, KVTVQVTV, VTVSKEKT, DVKVSVKV, NVNTNVNK, TNTNTNVN, NTVTVTVT, VNTNTVEN, TTTISIEI, RGTVSVTV, VTIQITIT, TLQVQITV, VTVSVLVT, LLLVSVEV, VTVSVTVE, TVRASVTV, QMGQGKVT, VTQQQGVQ, KTKSKTKT, NKTKSKTK, VTVTVTNT, TSKATVTV, ATATVTVT, VGTATVLV, IEITIEIT, DKTETIRI, NTYSYKYE, KNTNSYEY, ITISINKT, EATVSITV, DIVVSVFV, KITVSVEV, VVVEVVVT, TVEVEVVV, TLTATATA, ATASATVK, VSEASATA, VRVSVVVK, VNVQIQIT, TVQVQVQV, KQKSIQIK, VKEKSEQK, TTTTTTNT, TSTTTTTT, VTLTVTIT, TITIAVTV, ISISISIT, NISISILI, VSVSVSVK, ELSVSVKV, NTNSNTNK, EITNSNTN,NKNSNKKV, VNVNSNEN, KTVSVTVT, KLTKSVTK, ITISITID, KVTVSITI, EKKSKLKT, TDKKSKEK, NVNSVLVK, VNENSNLN, TVEVTVTV, ITVSVTVE, KVTVSVKI, YTYSYTYT, NKSHSYTS, VTVSVNIT, TKTVSVTV, TVEVSVTV, ITISITVI, NVTISIQI, VEVSVEIE, TVKVSVKV, FQFKFEVT, WVKFKFEL, EVTVTVTV, MRMEMRMR, EVEMEMRM, VTVAVEVT, TVRVAVEV, VSVEVSVK, EVEVEVSV, VTVEVTVT, ELRVEVTV, TAEVEVTV, KSKSKSKT, LDTDSKSE, FVFTFVFT, TLTFTFVF, VTVVSVVV, VVVSVVLT, TLSVSVVV, VVTVTVSV, KKTKAVTV, RTRSRTRT, TETESRTR, KDKSEDEV, VKTKSEKK, EVRVSVTV, TATASATA, LVTVTVTV, ITISVTVT, NITISITA, HKHKHKIT, TTVHKHEH, ITIQITII, VLTIQITI, ITIAITIV, ETKVAVTI, VEVTVTVT, VQTETVTV, IIISIIIK, SISISIII, VSVKVEVE, LIKHKVLV, TQSHSLTL, LELKLELT, VTELKVEL, KTKSKTET, IKEKSKTK, LTLSVTVT, VLTLSLTL, KTKTNTVT, KKTKTKTK, AVASAVVV, DVTVSAVA, FSFSFSFT, TLSFSFSF, TKTATAKA, VVVSVVVI, IVTVSITI, NTNSNTNT, TNTNSNTN, VTVSVVVT, VDKESVTV, LTLSLTVT, LITLSLTL, VTISITIE, TARVSITI, KQTISITI, QVTVSVTV, ATATATVT, IVTATATA, TVSVSVLV, VLVSILIL, VVVVSVLV, VTVSVTAT, TLTVSVYA, KVEVKVTL, KTKSITIT, VKAKSKTK, VVEVSVSV, VAVAVAVE, VTKVAVAV, TLTGSLSL, TITVSVKV, ATASFTFT, KATASATA, VTVSYTYT, NKSTSVIV, TLKVKVTV, KTKSKTIT, KKTKSKTK, LTLSLTLL, VRILSLTL, TVTVSVVV, TASVTVTV, DVDSDRVV, RREDSDVD, VTVAYTYT, ERRVAVTV, EEDSDEDT, DTIHSAKT, LLKVKVEV, VKVSEKEE, LKKESVKV, YTYSYTYK, VKEYSYTY, AIFTFIFT, IATATFMA, ITVSVVVT, VTVTDTDT, ERKVSVKA, VVVTIIIT, EVTVTVVV, HTHTHTLT, VNTHTHTH, VVVSVTVT, VITISVTV, NTKSKTKT, VDVNSKTN, TVSVSVTV, ITISITVQ, TTTISITI, TKTVTVTV, NNNKNEVE, NYKQKNIN, VTVKFTFT, RAEVKVTV, VTLTLTLV, TGTLTLTL, TAVTTVTI, RVEEIVIR, EREREEVR, KTDSDTVT, KKQKSKTK, KTTITLTY, VTVMVTVV, VKMVMVTV, DDTKSKTK, KKTKTVTV, ISISISIV, VVSVSISI, ILKVSVTV, VTATATAT, STSQSTVT, TTQSQSTS, HTHSNTNT, THTHSHTH, TVTATATA, KTTVSVTV, VYKVSVSV, NIVNKSKT, VENNNVSN, KTKSYKYT, NKQKSKEK, VQVSVQVT, ILQVSVQV, IEISIEIK, LITISIKI, NTNENTLT, TAENENTN, VATASATA, IVITITIT, SATITITI, KTKTITIT, IKEKTKTK, ETETVRVT, VLTLTFER, ITISRTRT, VNSISITI, NITVTVTV, ITIQITIT, TTQVQITI, VTVSITVV, VVVVSVTV, VISISITI, IITITITI, TTTSTTTT, TLTTSTTT, EIKISIKI, TEVSVSVT, VLEVSVSV, STSSVTVT, TLTSSITS, VTVTVEVK, VVTVTVKV, VVTVTITI, TVTVSITI, VTVQVTET, TRTVQVTV, ATASIEIK, EVTASVRA, HTHSHTVE, LSTHSHTH, IDISIDIT, VITISIKQ, TLTVTITI, VTVSVTVI, VFVVSVTV, TTTSITIT, TVTTSTTT, TVVTVVVK, EVTTTVVT, LVEVEVTV, IVTVSVTV, FTFSITIT, VKTISFTF, VSVSVSLK, TLEVSVSV, IEISIELE, KLKISIEI, TVNVSVTV, DTTVTVVV, VLVSILIT, TTSASVLV, TVTVTVKV, AVASARAT, TVEASAEA, ITITIDIT, TSTITIQI, IVISIVIT, KIEISIVI, VAVSAVAT, RLSLSVEV, TTTLTLTL, TVTQTVTA, VKVSVKVK, TIEVEVKV, ETEAETET, ENTEVEDE, FTFSFTVT, TLTFTFTF, IRIAIRVE, IIRIAIDI, TGEVKVTV, VKIEIKIK, IKEVEIKI, KLTVSVTV, VTVAVLVT, VVTVAVLV, AVTVTVTV, KTKTKTKT, IDTDTDTD, ESESKSKT, TTTESESE, TETSTETE, DLRTSTET, TVTVTVTA, NLNSNLNT, VTQNSNLN, TFSLSLTL, TTTSVSVT, TISTSTET, VTVSVTLT, TGAVSVTV, VTVRVTIT, DVEVRVTV, QITVTVTV, VEVTIEIT, VIKVTVKV, LVLTIVIE, RLTLTLVL, VITITITI, VLVSVLIL, VVSVSVLV, NNTNTVDV, VLTVTVVY, VNVTVTVT, RATVTVTV, NLVVSVTV, TETTTEVT, TLTTTTKT, ITVMVTLV, VAMVMVTV,VGTVTVTV, ELTISITI, NDSDSVIV, RITVSVTV, VGTASVEV, KKKSLKLT, KKEKSKEK, VTVQITIT, VLKVQVTV, IKISIKVT, EIKISIEI, VTVSRTRR, VDEVSVTV, ITITVTVT, TITITIII, NTNTNTTT, MKKNTNKN, KVTVSVTV, TETRSRTR, KVTTSVNV, RAKVSVTV, INITINIK, KLTVAVTV, TVTRSVTV, KSKTKSKT, TLTKTKSK, TTTVTVVA, VEVRVVVT, RLEVRVEV, VISISIQI, VTVTVIVV, DVDSTKIE, VVTDSVVD, TVTTLVLT, TLTTTTVT, EIIVSVSV, ETETETIV, TKIKTETE, RLTVSVEV, RVNVSVTV, YVYRYEYT, RVTYRYEY, QTQSQTQT, TETQSQTQ, VTVTVTAT, TFTVTVTV, INININIT, KINININI, VVVQVVVV, VAFVQVVV, VTVTVTQT, NNTVTVTV, HTHKHTHE, LKKEKHIH, KTKAKTKT, KKEEAKTK, VTLTLTLK, KTKEKTET, TKKKEKTK, EVTLTLTL, FTFTFTVT, VLTFTFTF, DISISITI, EATVSVVV, NTNSNTQT, TLTISITI, VSVAISIT, VVTVAVSV, IKIEIKIK, EKKIEIKI, NVNTYVVV, VKVNTYVN, NTNTNTVT, TVTTTNTN, TTVTVTVT, NTTTTVTT, VSVISITI, PTKTKTKT, VKTDTDTD, ATA I I I I I, TATATATA, YTYTYNVT, VKTKTYNY, TVTTSVTV, VKEVSVTV, SVTVSVVV, VATVTVTV, DIDSHIVK, VDVDSDID, RVEVTLTV, TETKTKTK, IINISITI, LVTVSVTV, KTKEITIE, VAKKKVTK, ITISVLVE, KLIISIKI, KLTVTLVV, VVSVSVSV, VIVSVIVT, VVSVSVIV, RLTVSVKV, ETYTYTYL, VKTETYTE, KITVSVTV, QTQSQEQR, IQDQSQTQ, TTTNTNTN, NVTVTVTV, VTSSVTVT, VTISIKIE, KVSVSVLV, ITITIKIT, DITITITI, VVVSVEVR, ELSVSVEV, EVKEKVKL, DRREEEVE, AIASAIIT, TASASAIA, VEVDVEIR, DLDVDVEV, VVVVSVVV, ISISISQT, TLTLSISI, ATYTYTVT, TLTLTYEY, KLTVTVTV, IGTGSITV, TLTVTVDV, RDEVSVLV, VTLSLTLT, VVEVSLTV, NVKVSVTV, DVNSNINT, ENVNSNVN, YVYSYVVV, IILYSYVY, ATAAVTVT, VVTLAVTA, IKITVVVK, EQTITVEI, VVVKVVVE, KVTVKVVV, TMVVTVTV, EIETEIET, TETETEIE, VQVSVVVT, RIQVSVVV, IEIKIEIT, VAKIKLEI, IKIEIKIE, RVKIEIKI, DTDTYTYT, TETDTDTD, DTDSKTKV, TDTDSDFD, VTVSITIK, EVTVSVTV, STSQVTVT, TSQSQSKS, VSVQVTVT, TITVQVTV, NTNSNTVT, RSTNSNIN, GTGEGTGR, ELEGEGKG, QTQTVTVT, VTTQTQTQ, TVTKSITI, ETKVSVKV, VTVSITIE, TLTVSVDV, VKVEVKVT, VKEVEVKV, FTFTVTVT, TVTFTFTF, VVVQVKVT, TVEVQVVV, SLSASAAA, ITISISVT, VDVRVQVV, VVTVRVDV, IKKESVSV, VTVAITVT, IRIEIRIL, RIEIEIRI, TVVVSVTV, ITVKVTVT, VVKVKVTV, ITIEVRVR, NIEIEIRI, YTYTYTYT, IATYTYTY, TLTITITI, TLTASVTV, RGTVTVTV, VTVSVTIT, ATATATLT, ATAQATAK, TGVAQATA, ATASATVE, RAIASATA, NTNTNTKT, TTVNTNTN, ELSVSIEV, AVASASLL, NAVASADA, VLTVSVKV, VVVSVTVE, ELKISVTI, ETESTTTG, EKKESENE, VTEVSVSV, VEVRVEVE, TVRVRVEV, ATASATAT, VLTASATA, VTSVSISI, LTLTLVLV, TTTTTLVL, TTSTSTTT, ITITITIA, RVTVTVTV, DVSVSVTV, VLTVSLTL, VTVSIQIK, VIEVSVTV, RVQVSVLV, VTVTLTLT, NTNSNQNT, VNRNSNLN, VVVVVVVV, LVVVVVVV, IEISIEIS, VLVSVLVA, TVLVSVLV, LQLSLQVT, RAQLSLQL, IEIAIEIK, EIVIAIEI, STSTSTSV, TTISTSTS, VTVVVTVT, VTMVVVHV, ASAQISIT, ALTAQASA, YVYSYVVT, RREESYVY, VAVSVSVT, VTLTSVSV, DTKTKTKT, VDTETKTK, RVVVSVEV, VVVVSVNV, TKTNTYTY, ATASGTGT, LAEASATA, ITITITVV, TVQVSVTV, VKVKVKVE, VTVTVIVT, VTVSITVT, AKAEAKAT, VVEVEAKA, GKGSGKVK, VLEGEGKG, AVASVVVR, DLTASVEA, EIKVKVEV, VTDSDTLT, TQKVSVIV, NENSNNLT, NAENSNNN, VTITIVIT, VVQVTVTV, RTEREVTV, KTKSIMIK, VKEKSKLK, LTLKLTLE, VLKLKLKA, TTTVTVRV, ITIKIEIT, TIKIKIEI, RTVVSVTV, VNVSVNVT, VVNGSVNV, VTVAVTVV, VVAVAVTV, LTVNVTVE, NLNVNVEV, EVKVSVVV,ITISITMT, TITVSITI, ATASVTVT, TVTASATA, IKTKSITI, HTHQHTHV, VKEKQHIH, ETFTFTFT, TGTETFIE, VRVAVRIE, RTTIAVRV, VVEVSVQV, QEQRQESV, IQTQRQEQ, IKISIKIE, NIKISIKI, VTSVSVNV, REVSVEVT, VRTESVEV, TVPVSVTV, KTKSKTVT, TKTKSGTK, DTDSDTVT, DNSNSDTD, YSYSISIR, EIIYSYSY, VKVEVKVK, RFEVEVEV, ITISILAT, KFTISILI, DLTVSVTV, TVTVTVIV, FTFSFTFR, ELTYSFTF, RTETETET, VQTRTEVR, VKVSVKVN, DLEVSVEV, ETESVTVL, IKSESVTE, TDTKTKTK, ITISITIR, DNTISITI, VQVQVQVT, RVQVQVQV, IKVSVKVK, VASASITI, VITVSVIV, TNTNTKTN, VVTVTLTV, VITTTVTG, KTKEKTLQ, TEKKEKTK, VLELSIKI, NVTVSVKV, NISDSVTL, ELTVTVTV, ATAAATVT, TATAAATA, VTVSATAV, VLAVSVTV, VEVSVEVE, TIHVSVEH, AVASAYAL, VVLASALA, KKEKSVTV, IKISIKIT, KVEISIKI, KNTNTNTN, LVLSLVLT, EIKVSLVL, HTHAHTHI, NKEKAHTH, VTLSLALK, DEDDDTIT, EKRDDDVD, YVYTYVYV, ERTYTYVY, FQFTFQFT, TATFTFQL, VTVEVTVR, EVDVEVRV, ETRARTRR, LDDEARTE, VGTGAVTV, RLTVTVTV, EKEEYKYV, KEEEEKKK, ITIKITIE, KVTIKIKI, VTVTVLVT, ALTVTVTV, EVTVTVKV, ITIEITIV, VVVIEIKI, KEEESKTK, MTMTMTVE, VVKVTMTM, VTVAVAVT, EVKVAVLV, GTGTGTGT, TLTGTGTG, DVDEVVVR, HTDDEVVD, ITIEITIR, ELEIEILI, VTVSVRVT, TVEVSVIV, RVKVKVIV, NVDSNVIT, VATVSDVD, TVTVQVTV, TLSVTVTV, VTVTITIE, TVKVTVTV, ITTISITI, AVATITIT, VVTATALA, ATASVTIT, TLTASVTA, KVTYTYTY, ITITIIIV, VVISIVIK, DVEVIIVV, ATTNSVTV, TESHSVEV, NIKVKVEA, KTKLKQEE, TKQKLKQK, TATVSVTV, VTLSVTVT, ALSVSVIV, TVTVSVEV, VSVSVTVT, RITVTVTV, VTVTFTFT, DKCKSYKY, ITISIEIT, TLTITIKI, TMKISITI, KVTVAVEV, ATLTLTLT, TATATLLA, IIISITIT, KIEISITI, TTTVAVAV, YTYTYTYE, KNTYTYTY, TTTTSITI, ELNVSVTV, VKVSIEIK, EATNSVSV, VIVISIKI, TTTKTTTT, EVKTKTTT, ATATATAV, RVTATANA, TQNKTYLL, RISVSVTV, VVVSLVLT, TLEVSVVV, ITIEITIK, ELTIEIII, ETESHTHT, DDTESEKE, DVTVTVTV, VIQVQLTV, YVYTYVVT, VVTYTYVY, TVTVTVLV, VTINVTVT, LVNVNVTV, VEVTVEVK, TVTVTVEV, LVLMLVLE, KLMLMLVL, EEETVLVR, ERTETELE, LQLSLQVE, RLTLSLQL, IKISIKII, VIVISIKI, NLTSSVSV, LTVSVTVR, TLELSVRL, VAVSVAVT, AIATVIVT, TVTATAIA, LTLSLTVE, LFKLSLTL, VTISITIT, RLSVSITV, TTTSTTVT, VKTTSTLT, NLEISITI, TLTVSITV, VTVKITIT, TVEVKVVV, ELTASVTV, KVEVQVTV, VEVSVLVT, VLEVSVLV, TKTKTVTV, DVDSEVEV, VDEDSDVD, IATLTLTL, LSVSVSVT, RITISVTV, VEVNVEVK, EVNVNVEV, TTTSVTVT, VTTTSTTT, VQVSVSVR, EVSVSVDV, RVEVSVQV, VKVTVKVT, LVTVTVKV, DVEVEVKV, DVVSVTVE, KKVKSKLD, LTETELEV, TSTETETL, VQLSLQLT, TLTVSVQV, QTRTRTRT, VDTQTQTQ, VATVQVTV, ITISILIE, IVKASALA, KVSVSVNV, VTVTVTVE, RLTVTVIV, LLEVSVTV, KTLTEVET, VTVGVTVT, TVTVGVTV, ETETVTVV, VKTETKTE, TVTVSVQV, KEKTKKVT, IKEKTKEK, HITVSVTV, AEASAEAV, RAEASAEA, VGTISISI, VITDSDKD, IVISIVIT, LLTISIKI, IDIAIDIT, VIKIAIDI, VQVAVQVV, RVEVAVQV, HEHRKTKT, RSESESIH, VEVAVLVL, VVKVAVLV, EKKDSVTV, YTYTATVT, LSLSISII, TLILSLNL, VEVSVEIT, TIIVSVEV, VATVSVTV, SSSSVSVT, TLTSSSSS, EAKVSVTV, ISVSVSVT, HTHSHTLV, DIKHSHTH, LALSLSLT, VLTLSLSL, LIKISITI, VKVEVKVE, ELKVEVKV, HKYSYDYK, VKTHSYHH, VTVAVTLE, VLRAAVIV, KIKSKVKV, VDVKSQIK, VLTVSITI, VEVSLELT, QLQVSVKV, ITISITLV, DITISITI, TTESETET, TTTTSLTG, VQTNSVTV, VTVSVTVR, NVDVSVTV, ELTASATA, VRVSKLTV, SNSELTET, TRRSELMV, AVLKLEER,TSVSTHKI, SESMHTQT, TEARMHGI, SVHTELTR, VLTNTYSI, STSTYTRT, TVKNTYTI, NTYTESVS, VDTLTYQN, STFTYTRK, TKTLTYSN, YTYTKTVK, VTKNQEAR, STSSEQKT, TEKKSETR, SSEQKLVK, VLTYTWTV, STSTWTLK, SETYTWNV, FTVTIAVK, ITEKRYAV, SEYQYVEK, TQTKQVNV, YTYRESTK, VVSVTVNL, ISSNVTIK, TVTYNVNL, VSVTVTVK, VLSYTFNV, YTYTFTVV, TNTITFNV, YSFTLTVR, VVTVTYNV, QVTVTYYD, STSTVTTK, TKTVTYSD, VTTTVTEK, TTTSTSSL, STTSTSDL, STSTVSRK, VVESTYNQ, TSSTKKVV, TEESTYNQ, KTYTKKQS, VVTVGVTV, TVKVTVNV, VTVGVSVK, INMTKVSV, SSTTVTNT, ANKTTVNV, TMVKNLVK, IVTNNYNN, SSNINEQK, SHTNIYNN, KNNKSSQK, VDTSTHTI, STSQVTRT, TETVQHSI, STHTRTVK, VETYTYSV, YNYTVSVV, SKTYTYTV, YTYTVHVK, VASFTYVV, KVTVTYVV, FNYTVTVK, VDKSEINI, HTSRVEQT, SVKSRINI, HKIEQFEK, VENVTYNV, VNVKITNT, RKTVKYNV, VSYTYTVR, KVQLQVTL, VTPGVETV, EIGTGVQA, VTVQPGTV and STLKFERT. The one or more beta strands and / or one or more alpha helices may comprise one or more of the listed insertions. Different combination of these eight amino acid elongations / insertions may be used in the same beta strand or alpha helix or in different combinations of two or more beta strands and / or two or more alpha helices. The eight amino acid elongations / insertions may be used in the one or more beta strands and / or one or more alpha helices consecutively or separated by existing sequences.

[0500] The one or more beta strands and / or one or more alpha helices may be elongated by one or more of TVTVTVTVTV, VTVTVTVTVT, KVTFTYTVTL, SEFEVEVTVT, SVTFKYEVSL, FTFTYTVSVT, TLNVGVGVTS, SSSSVNVLVV, SVTVNVNVTS, NNVGVGFSVT, VNTVTVEGRG, VNVRVEVTRR, SVEVRVEGEG, VTVEVTVRRT, TEVTENNYTA, TTNSYKVETT, TKRNENQYSA, TTNNYKVTTT, VLTATVTNTQ, STSTTTVTRT, GETVTVTNTQ, ATATVTRVVT, ETSITITYSL, STTTITINVK, STTTEITYNL, ISITYTYKRT, RVRLIVTYSI, SEVTVEVLRQ, SEELEVEYNI, LRVTVLVQQT, VLSISVSYTA, SSINYTVTNT, TTTITVTYNA, ISISVTVTQT, RVVVTVVYSV, STITYEVNVQ, TVTVEVEYNV, VTVVYKVTVT, TNTNTNTFNS, NNNTNTNTTT, TNVNTNTFNS, NTNTNSNRET, IDLVEIEVTL, SNVRVRQNTK, SRSVEIRVNL, VLVEIMHSVT, VATSETTVSL, STSTQKQTYR, SRRLETKVEL, STIEQKYERT, VFNNKYEYTQ, SNSEYEYTER, SRSEKYEYEQ, NKVHYEYAET, VTTVMNKNNT, SNTELEVTTR, TRTVENENNT, VTNKYTVQET, VLTVRVEYRV, STVAVEVTIE, TLTVEVEYNV, VTVEVLVSQT, VQINENKNRN, SNNTNEYELV, SRSNEMENNN, SINKNEYKVT, VTTVTVTVTS, STVTVTVTRK, STKVTVTVDS, VTVTVTTKVT, TLTIEITISL, STSEIEITTT, TVTISAEINL, ITITITTQVT, KISVTVTYTL, SNVTVTVTVV, SVTVTVTYKL, VNVTVTITVT, VTKNTNTDNH, SQSTDTNEKK, STTNTNTDNH, NTNTVTETVT, VVEVTVSVKL, SEVTVTVKTT, TVSVNVTVKL, VEVSVKVSVT, VENVKIKYNV, STSEYEYNLS, RKTHEIEYNV, VNIKYKYTQT, VVEIEVEYMV, IKQRIKVKKK, SNEIRVKYVV, IEVEYKVKET, VTEVETSKTA, STSSKSTRTI, TKSVETSKTA, VTTSRRTTVT, TTTLKLTYNA, GTLSYEYNKT, SDTLELEYNA, VSLKYTNATT, VTQTVVTYTD, SSSSVSVVNK, STTTSVSYTD, TITTVVVNTT, IREVVYEYTY, VSVKVEEKEV, TREVKVEYTY, VKVEYKETVT, VDKRISIITD, STIQIQMRTV, SKTRQSQIND, ITIISISSTT, VLTVTITISN, STSTITKTVK, STTVTITINN, VTITITKKVT, VTEVNVRYTH, SSSEYRVIVR,TRTVTVRYQH, VEVDVNVTVT, VLQVTVTRTR, STLTITNNTT, KGTVTVTRNR, VQVTVTNTTT, INTNTTTVNV, VTTEVEYKQV, SQSNETEVNV, VNTTVNESTT, VANVSVEYRN, STSRVTVTTK, TVTVTVTYNN, VNVEVTVTVT, VLTLQIQINV, SSITIIVTVR, TKTLIIIINV, ITIQITVTVT, TGMVTASDND, SNSTDAATVT, TTKVAAADND, STASATLSTT, VVTVTVTVTV, STVTITVKKT, RVTVTVTINV, VTVTVTKSVT, TSTITITYTK, SSITITTTVK, SVTIEITYTK, ITITITTKVT, RQTILVNITV, SNSTITFSRT, SRKIIVTISV, ITITVSFSET, RIEITVTYTN, SGIVIVVTNV, RREIVVVYTN, ITITLSVTMT, VKTVTTTYTV, STRSYSRTVK, SETKSISYTV, VTVTYTESRT, TVSVKVLVRN, SEVEVEVKIT, SVTVEVEVNN, VSVKVKISVT, LTKHTSIYQD, HTSTKEVEEK, SQKNESEYND, HTEIKIESVT, VTTVTVTKTN, STSTATVTRK, TVTVTVTKSN, VTVTKTVKST, VKSTLVAVRV, SGVEVEDTTI, RTKREVEVRV, TSVAVTDSVT, VSEVTTTANV, SDVVVKNKTK, SKEVTTKATV, VEVTATNSAT, VFTITITYSN, STSTITITTK, TRTRTDTYTN, ITITITITRT, VYTTQTQVNQ, STSSVEVKET, SVETETEVNQ, SNTQVKVKET, VNKVSQTYSN, SSVEQKEELT, SQRVKVKYNN, VEVTQEEQVT, VTTQTQTIRT, STSTISQTTQ, TNTQSQSIST, STQTITQQST, TTTVTVTIKV, SESTITVTVN, SVTVEVTITV, VTVTIKVSVT, VKTVTTNDIK, STRSDEETEK, SRTVTTSDNK, VTTNDTESET, ITSVTVTYTL, STSSVSVTRQ, SETVSVSYTL, VSVTVTVSQT, KETVLTVVNV, LNTQVANNTV, SKTVTTAVNV, VTTVATNTTT, VNTVTVTYSN, SNSSVTYTQT, KETVTVTYTN, VTVTVTHSVT, TTLNTYTYQL, SKASYSLERT, SERNEYSYNL, NLATYTQLRT, VNTNTNTNKN, NESTNTNTQT, ITTTTTNTNKN, NVNTNTNKVT, VKQYQYRATT, SNYEYQYTQV, RQTYVYQANT, YQYQYKQQVT, VVKTTITDTD, SSSRITVTVE, STRTEVTDTD, TKTTISVSVT, ITTYTYTYSN, SNYVYTLTTK, TLTVTVVYNN, YTYTYTLSVT, TRTVSVNNSQ, SSYSYNVTQT, TTKTSVNNNQ, KSVNVTVQST, IKEITFTYNQ, STSTYTEKII, SKEITFTYNQ, VTFTYKEKIT, VTTVSVNYSL, SNSKVEVTVT, TKKVEVEYSL, VTINVSVSTT, IEKITNTNTV, YTSTNTNTTN, KVTKTETNNV, YKITNLDTST, VQIIVYVYSN, SKIEYEKKVT, KETIIYEYDN, ITIVYKKTVT, VDVTEKEVSN, TKNETEKRNV, TDTVEKEVEN, TVKEKRKQVT, VLKVTINVSN, SNITVKVEVT, TVKVKIKVNN, LTITVELQVT, VLEVTVTVSL, STVSFTYRKV, SNEVTVTVDL, VTVTVRYKVT, IDKSISIRTD, SNSERTDITR, SRESTTTRND, SKSIRIDKRT, ISTVTISYTV, VSVTYTVTTV, RKTVSITYNV, VTASYTTTTT, VQEVTYSYST, QNTTYTYREK, TRTETYTYNT, VNYTYTYTVT, NVTIKIEVNV, IGSTIEVKVN, RVTIEIEINV, ITIEVKVTVT, TVTILISYSV, STIEYKITLK, SVKIEIKYNV, ITISITIKET, VLTLTVTATL, SSITVSVRTT, TKTVEVSATL, LTVTVTMTVT, VRKNTNTYTL, STNKTENVRK, STRNENEYTL, NTNTDVVTQT, VSSNVNTVRN, SENTVTENEV, TKTRTETVNN, NTNTNTETVT, VNKKEDTDTH, DTNEDIDETT, KQEKRDIDTH, DKDEDESEVT, VDSSTSTSTL, STSTSTNTRN, SETSTNTSTL, SSSTSTRAVT, VTTVTVIYNV, STSIYTVNET, SKSVTVTYNV, YTVIYNVKST, QQLLLLTYSV, STIQIQQQQQ, TQWLQLQYNV, LRLVYQQQVT, VARHTYTKNN, STSTKEVETR, TTRTEYEKNN, HTYTYVVETT, VSKITNEYTA, SNSTYTNNVT, KNTITNTYNA, YKIENTNTVT, KVTISVTLNL, STITFTFTVI, SRTITVTLNL, ITITFTSSVT, VTTISLNLSL, SNSELELTRR, SEKITLELNL, ITLNLSTHQT, TTINININTV, NNSKNKVKVT, SVTNENKNLV, NININKVSVT, QTVITIAVNK, SDITVTVEQK, TNTVTITVNK, ITITVTVQVT, VQQNSNKDTV, SSSENRNELQ, TTRNENRDTV, NDNKNLNQQT, VTTVVVTYNY, TLVTVVNTVV, TTTVVVVYSY, VTVTYTNQVT, ELTVEIRVSV, VEVEVVINVK, SVTVVIVVNV, VTIRIEVKVT,KVTVVNVNTI, STVSNAVTVT, SVTVANANTI, VTNVNTVSVT, VDTETKTINV, ATVTVVKTVT, SEKEVKVINV, VTVTKTKQVT, VNVDERVLSL, SNNQKQQEQV, RTTDRRQLSL, DVPVQEQTVT, VIKITITVKL, SESKIEIEKK, STKVKIEVSL, SKITIEIQRT, VDKKEERYND, STHEYIDENT, SEKKKEIYND, KKKEYEDINT, IKKKEVAKKL, SEIEKEVEET, TKKKEVEKEL, KEIKKEETVT, VKTVTINITL, SNVTIVATTS, SQNVVIVINL, VTVNISERTT, VLTLTVTLSA, TTVTLTVTTT, TRQVSVTLTA, LTVSVTVRLT, VVEVTVNYNV, SNVTVTTKLT, TLEVEVTYNV, VTVNVTTRVT, TVTVTVTYTE, STVTVTVIIR, STTVTVTYTE, VTVTVKVSVT, EVSVTTTVTV, VTVTVTQTDK, SQTLTTTVNV, VSVTTTLTVT, VAEIEININV, SNSKIRIKVT, TRSIEIRINV, IETNIKVTVT, VQVIRVTVDV, SKVSVTVVQR, TQEITVTVNV, IVVTVRQQRT, VTIVTVTVNN, STVTVTHRVT, TRTETVTVNN, VTVTVERSVT, VLSISISINV, SNISISIKKV, TVTISVSINV, ISISITKTVT, VITEEKSNTV, SSSTNKITIK, SKTEEKENNV, ETKKNKISET, VRRNTNTNTI, STNENEEEVR, TVENENENII, SRNTNKEQRT, VFVSSYSYTN, SSSTYTYEVK, SATSSYTYDN, SVYSYSLSVT, VDTSKKTITY, STSTIDTTRV, SVTSTKDISY, STKTIQTTVT, TSTVTVTVTV, STVTVTVTVS, SVTVTVTVTV, LTVTVEVSVT, VTVSEFNYNL, STSRYRYETV, SRTSRFRYNL, STFEYEYRST, KEENNTNYTH, SNSNQNNVVN, SVTNNTNYNH, NNTNNKNSVT, VATIVVATTN, SNITIKVQRK, STTIKVKTSN, ITVVFQVSRT, SATYVYVRKL, STSEYEYTVN, TKTYTYERTL, YKYVYTYTST, VRETKTEISL, STSTTTNKEV, SNTATNTINI, TEIETKESVT, VTVVTITYTN, TQVTVTVKVV, AVTVVVTYNN, VVVTIKLQVT, KRNTEIKISA, SNSEIELETK, TKKTRIEINA, TNIKIELNET, VETETATYTD, STSTYEVTRK, TTKEEVEYTD, ETVTYTVEST, IEEISFNYNG, SSSEIENTRR, TEKIEFEYNG, FDFRYRNQRT, VETRRVTTRE, RESEIEETTT, TRTREVETTE, RTVTVRESVT, VLEVRTKVRD, SETEVEYKVT, TKEVETEVDD, VETKVKYKVT, EVQLQYQYTL, STSTVTIQVK, SVTLTNTYTL, LQYQVQVRVT, VTTITITITI, SGLNITHTRK, TETITITINI, ITITITRTKT, VLTDTVTYTD, STSTVTRTVR, KETDTVTYND, HDVTVTRSVT, VLVVKVDYKR, SNVVVEVTVT, SVRVEVEYDR, VVVKVTVQVT, VNTTIINYNV, SSSTYEDTRV, TNRTEIEYNQ, TTIKYTNRVT, VDTVRVEVRV, SSTVVVTTKK, STTKVVVVNV, VNVEVTSHVT, VDTVVVKVSV, VSVVVVQTKR, SDTVVVVVNV, VTVKVVQKVT, VATVQVTTTS, STVTTIVSLR, SLSVIVTTNS, VTVQYSVSRT, VNTFTFTYEL, SKSTFTNEVT, SKTFTTTYTL, FTFTFERSET, IQVHTNTYKV, SNTTVTNETV, SNTVTNTYNV, HTNTYTNSVT, TTEKSYSVKV, SNYTVSEKRT, TENKEESVNV, SEYSVKEKRT, VLKVIVTVTV, VTATLTTTVR, TVTVTVTLDV, VKVTVTTTVT, VRTNANAVSV, STSDVKVTVR, TETNKNKVEV, NTNAVTVTRT, IEKIEYSYSI, SSFSYSNEVK, SVKISYSYNI, IKINYERSQT, VATVNYNYNV, SLFTYQLQTR, SNTVQYQYNV, VTYNYTLSTT, VYTNTTTVTI, STTTVSFTLR, SYTNSTSVNI, NTTTVTFSST, VNKNVKESRE, SQSKDEKETK, SQTNEKESNE, NVKEKKVSVT, IYKVEIKISN, SEVEIEIEIT, TIKVVIEINN, VKIKFEIKRT, TVKVTVTVKL, VESSVTVTET, TRSVTYTVNL, VKVTVTVTET, VDKHIHTYKV, TEHTYTREQR, TERDTHTYTV, HTHTHEREVT, IYRYKFEYSV, YTYEYEYKTK, SVTLEVEYNV, YRYEFKYSET, KSTIQITVNV, LTIISINDQT, STKTIIIVNV, ITIQNDNQVT, QVTVTVTYTL, STSTVTVTEN, TKTVTATYTL, VTVTVTVKET, INSVSVEVSD, VKVEVKVEVR, SVPVKVKVSI, VSVEVETQIT, IETVTITVNN, SNITVTVTKK, TETVTITVNN, VTITVTKKVT, QNTQESLTTY, QSSETKNNRV, KQTKKSKTNY, QTSLSENSVT, STETETTTNV, SKTKIKNNIT, TNETKTKTNV, TETTTKNKVT, VERNTNTDKV, HNNTDINERV,TTTNTNIDTV, NTNTITSTET, VEEYEYKHTV, STSKHKKKLK, TEEDKYKHTV, YEYEYEKSVT, VVTTTVVQTH, STRTQTVMTA, TRSTTVTQTH, TTVVQTVTQT, I I I I I NTFTN, STSTNTVTRK, SIIIIII FNN, TTNTNTVSVT, TVVVTNTNTV, STSTNTVEVQ, SVTVTVTNNV, SVVTNTIKVT, VVTVTVTVNL, STVTVTVTQT, SKSYTVTVDL, VTVTVKVQRT, VVVETVTYND, SESRVTVKTT, TRTVEVTYND, EVITVKVTET, VEEEKKEYTV, SSNKEEKKKV, TDEEEKEYTV, EKKEKKKKVT, KNTITLKVSV, STITVENTET, TKKVELEVNV, ITLKLTEEVT, KSKYEYEYTI, YTFKYQYEIT, TTKYQYEYNI, YNYKYEYEVT, VNSNLNIRQN, SQNENRNTIT, TAKNENRRDN, TVNINTVQTT, VSTVTVTYDA, SNVEYEVTKK, STKVEVEYDA, VTVTYTLSVT, VDQTQVNIRI, STTIIIVTQT, ATRTEVIINI, AQVNIVSQTT, VKTVENSYNK, VNEKNEVVTS, TLTVENEYNK, VLESNKVTRT, RAQFVFTVTV, STFTFTFVVR, SETRTFTVTV, FQFNFVLAVT, TQQLTLTLTN, SNLSLTLTRT, KTTTTLTLNN, LTLTLTLSVT, QEERSYRVRV, YSREVKHHRR, ADTREVKVDV, REYSYRHSVT, VLSTEVNVTV, SNSQVQYTVK, TRTVQVQVNV, SSVEVVNKIT, KKTNTNTSTL, SSKTNTNTTT, NKTETNTSDL, NTNTNINSTT, KREVEVTYTF, STLSVKVNIT, KVSVKVKLNF, VEISVNVTVT, VTTNSNSYSI, VNLTDVNTIT, TESKVNVYSI, VTNSNSNTTT, VVTQTQDYNN, SNQTVTQTTR, TRTEVQTYNN, QTQTQTVEET, VFLSTYTYTD, STSTYTVRKS, SVTVTYTYDD, SIYTYTVSVT, VNTTISTNNQ, SASTNKKTVV, KETAVSKNNQ, TTVTKINTVT, VNQFTYQYTI, STSQVQYTKK, TNTFTVQYTI, FTFQYTYTTT, IDEVEVEYMA, ITSSYSAVTT, TKRVNVSYGA, VEVEYTHVTT, TRRGVITYKA, SESEIENVEV, KRTKRIEYTA, GTIVYVETVT, VVTTTITITF, SSVTITVTVK, SVSTTITINF, TTVTISVSVT, VSTVTSSYQV, YNATYTITQI, KNTVLSTYDV, YTLSSTITVT, IVEVKVTVTN, STVTVEVKQV, TTTVEVEVNN, VTVTVKVQET, VYKSQVQVRI, SDVSTEVEQT, TKKTVVSVGI, SKYQVEVNTT, VVAVTITISI, TTSSISITRQ, TTKVTISITI, VAITITIQQT, VTNKTNTNRQ, SNSQNQIQQT, TTVKQNQNMQ, KNNQNQQRTT, VEEYISSSRI, SNSTNRSRRR, TVETRSSSNI, YTSIHISKRT, VNTVTTTVTI, ITSTVTNTQK, TNTVTTTVNI, V I I I I I NQIT, VNTYSYSYSR, SSAEYRKEAT, TTKESYRYNR, YTYSYEKLQT, TVKTELTLAY, TTSTLQYTQK, TQTTRLQLTY, TTLSLEYTKT, VKTKKYNYSL, SNKEYENTTR, CKRKENEYNL, KTLNYTNTTT, TTTVTVTVNV, VNVTVTNTVR, AKTVTVTVNV, VTVTVTVTTT, IERNEHKKTE, SNSQHVKEKR, SEKQVNEKNE, NINKHEKQVT, VANHIHRVQN, SESEHRHIQQ, NETEEHRVTN, HEHTHIQSTT, TVTVTVTYSE, STVQVQVTLT, SQSIQVQYNE, VTVTVTVQVT, TFNNEVSVTV, VNSNVNVTVK, RNTNNVNVNV, NNVSVTVTVT, VTTKTETKNV, STNTKVVNVK, SVTQVEVKNV, KTNEEKVSLT, TVTVTTTVTV, STSTQTVTVR, TVTVTTTVSV, V I I I I I VTRT, VATVTVTVNV, VNVTVTVTTK, TRTTTVTVNV, VTITVTTTET, RDMVKSKVQI, SESEVETVTT, NVKTESEVNI, VYVKKKTETT, TVTVTVTVSV, STVSNTVTRN, SVNVNVTVTV, VTVTVIVSVT, VNRVQYNYNL, SDYSVSVTVV, HTTVENSYNL, VRYQYTVSVT, QVTVEIRYSA, FSLTVTVNQQ, HTTVTITYTA, VTIRVNIDQT, VKTITVTVTN, STTTVTYTEK, TLTTTSTVTN, ITVTVIYKET, VRTVIVQVEY, STSQVVVTKT, TVETVVVVNY, VTVTVKVRVT, VRTITYTLKD, STTTLTQSEK, TKTETYTLQD, STVTYTDSVT, TSTNSNSNTN, SNSNNENTVT, SIAKENENNN, NTNSNSQRLT, VVKVKFQLNV, SDSTLEVEVK, SVKVEKTLNV, VKVQFAVELT, VNEYKYTYTV, YTYEYKYQQK, TVDHKYKYSV, YEYTYALSQT, VLQINININV, SSISISITLT, TRNISISIGV, IQINISLTVT, NTNIKQTIRV, SESEIEINRT, SENIEQEINV, INQTQKISRT, KKNINPNISI, SSININNNIT, NKNTNVNITI, ITININNSVT,INTVTVTVNV, STLTVEVNKK, KNTKNVEVNV, VTVTVTNTET, IVINRNTYRD, SENTYKNSKT, TNKNQNKYTD, NSNEYEVQVT, ERSLRLTITI, SNLDLEVSVT, RVVLELELTI, LSLTLRVTET, KTEKTETVTV, STQTVSKTVV, TESKSKSVSV, KEKTVSVTVT, VVEFEYSYNI, STSSYKWVVT, KQEFRYSYNI, FEFSYEVRVT, VRVDENNKSN, SNSSKKTTES, SKSDKDKKNN, DIDEKSTSVT, VDRVKVTYTV, STIEYEVTRT, TEEVEVEYTV, VRVTKLVKRT, RLNTKVTYKV, SDVVVEVIVT, TVETKVEYTV, TTVTYELKVT, VTTYTNTITV, YRYENENRVV, STESENEITV, YTYTNTHRVT, VEKVTVSVNN, VNVTVTEETT, SRTVTVTVSN, VTVTVEERTT, VLTVPVDVNV, SNVVVVVTRV, STTNVVVVNV, VKVDVTEKTT, VNCVTSTNTN, SNSTNTKTQQ, SKNVTSTNTN, STVTNTQSVT, VVTQTKTYSD, SNVTQTLTLT, KKTIKLTYSD, QTVTKTLSTT, NLTVTINYNV, SDVTVTVTLT, KVTVTITYNV, VTVNVTLSVT, VVTVNSSLSL, SSVTLTVTEA, RLSVQSTLNL, VTVSVSVTRT, VETVTVTVTT, NRVEVVKTRT, KETVVKTVTT, VTVTVTTQRT, TITVTNTSQV, VRRTVTNTVK, SVTVTNTSSV, VTNTVTVSVT, TVLSQYQNDL, SNSVNVHQEV, TRTSVYVNDL, SLYQNQLTVT, VNDASDLDSD, SSQEDHQRRQ, SETATQHDSD, QSALDTQSRT, VVEIEVTVNL, STIIVIVKAK, TKEIIVIVNL, IEVTVKVTVT, IVSVQVIVNV, ITVNVNVTIT, TVSVNVNVDV, VSVQVTVVTT, VTQISQSQNV, SSSSQSQTQQ, TTEINQSQNV, IQISQKQKQT, VNENVNTYKN, SESTNTVKII, TTTNTNTYKN, NENTYTVKVT, VNENTNTNTN, HTNNNSVTKK, TETNSQSNTN, NTNTNTVKVT, VVTFSFTYKQ, STSTYTYVER, KRTIKFTYDQ, LTFTYVETRT, TIESTYTYTN, STSTFTLVVV, SVSLTYTYTN, STYTYKLGVT, INKITSNITI, SSSKIENNNT, NTKIESEINI, ISINNTNEVT, KLTITVTVNN, SNITVTVTQT, STKITVTVNN, ITVTVELQET, TDTVSVSVST, SSVTVTVTRR, SESVTVTVST, VTVSTSRSTT, VNTYEYEVNV, SNFKVKEEVK, TKTVEYKVNV, YTYEYKETLT, VVTVTYNYRT, STVTYTVTVK, STTVTVTYNT, VSVNYTVSVT, IRKISYKYTL, SSSSYEYRET, SRRIEYEYSL, ISIKYSYSET, VVTVEVEVTE, SSKKVKVEVR, HVSVKVKVTE, VTQEVHVEVT, KVEVITWINV, SNVNINVAVN, NVKVETNINV, VNTNTAVETT, VVKVVVVVNL, AQVKVEVEVT, STKVKVEVNL, VKVVVEVQQT, ETTNTVTVTV, SQTTITNTVK, SETHTNTVNV, ITTTVTRKET, VVTVSYNQRQ, SQSNQNVTVQ, TVTVNYNQNQ, VTVNQTVTVT, VVTVTQETQQ, SNSSTSITVK, SVTVSQSTTQ, VTLETTVSVT, IDEHLKEVRV, SLSKVELRRR, KETHIDKVSV, HISEKRLTRT, VSTSTYTYSI, SNSSYSKTHR, TQTSVYSYNI, STSTYTRTVT, VQKQENNVSV, SNQEQENTTV, STTQGNEVNV, QKQNNETSVT, VLKIPVSYNT, SKSSYSNVNT, SKTISVSYTT, IKVGYVNSTT, VDKVKYVYNL, SRSEYEEERR, TETVEYEYNL, VKVVYEHTRT, VIRVEVDITL, SNVEIEVERT, SVRVEVEIDL, TRVEVEVSTT, VEKVSVEVEV, FKSKVSVQQN, TKKKSVSVEV, VKVEVSQSIT, TEKNTNTGSN, STNTNSKTKT, STSNTTSGGN, NTNTLSKTVT, TDRSVSMNTN, SSSLNNKTKV, SETSNKDNEN, SRSCNVQSVT, TVTVSVSFTA, STATVTVTVT, TQTVTVTFTA, VTVSVTVKVT, TVTVVVTYNE, SVTVNVTYTE, VTVVVTVKFT, INRYTYNYQN, SKSSYTNEVR, STAYTYTYQN, YTYNYTNQVT, VIKEEDTTIV, VTEETKKETV, KTKEVDETGV, EKDKKEQEVT, VLTITVTYTD, SSITVTVTRR, STSITVTYTD, ITITVHIQRT, VTTTSHSNKN, SNSTNSNTLT, SKTTSHSNNN, SDTSYTLSET, VTTVTVTYTF, SSVTVTVTTK, TLTVTVTYTF, VTVTVTVKKT, MNTTTITYTV, SQFNYNNNVR, SNTTTINYTV, FTITYTNSTT, VLTVTVTVNT, NTVSVTVTKT, TTTYTNTVTT, VTVTVTKTVT, VNTQTQTNNV, STQTNSVTVT, SVTQSQSNNV, SHQTQTVTTT, VVRSEFTSRV, SESESRTEVT, STTVRFRSNV, SEFTFTTSIT, TRTTEVTYNN, SNNTVTVTEK, SRTTTVTYNN,TTVTVTVSVT, VDKSSHSLDD, STSTLTHKVV, SETTEHTLRD, STHSYSHTTT, KKQVTITYSV, SRVEYEVTIT, SVRVEVEYNV, VQVTITVSTT, IDDYTFTLSN, STYELERRVT, RETYEFELNN, YDFTLKRTVT, VEKFSREDTQ, FNSTDTDTQT, TRKTSRTDTQ, FTSEDTVQET, VDTVTVTVSV, STVTVTHTEK, SKTVTVTVTV, VTVTVTHSET, VYRYSVKYSV, FTYSYEVTIL, TVTIEVEYNV, YAYKYTVQRT, VLTTTYSYQN, SNVTVTNTNK, TNTTTVTYNN, TTVSYTNTVT, VSREEIIISV, STVEIEIEDR, SKEEEIEINV, ISIEIEIKET, VEKKRKETEK, SKSQYEQKTE, KETKEKETEK, KKEEYKLTKT, ILTVTVTVTL, STVTVTVTTK, SKTVTLTVNL, VTVTVTTKLT, VTRSEFSYTE, SNSTFTYVQN, SQKRTLTYTE, SRFEFEYQST, VETVTVEYSN, STVTVKITRK, SETVKVKYNN, VTVEYTHQRT, VDVHTHTISL, HTHTHTHTEV, KETHAHTINI, HTHTHTHTVT, VSENTDTDTD, NTNTDTDKIK, SKENTDTDTD, NNDTDKDKVT, VVETKHEYSV, STNENTHKVV, RKDNKHTYTV, TKHEEKVTET, VEKSRNSNIN, SNDENENTTT, TNSSENENNN, SKNSNKITTT, VKTNTNTYTN, STSKVKNTTV, SDTKKNKYNN, NTNTNEVKVT, LKKVTTTVNV, SNVTVTEEET, MKKKTTTVSV, VKVTTTEKTT, ILTLKLTLSV, SELELELTVT, TVDLQLELAV, LTLTLKLQVT, IETLVYIAPA, LKVTVEYTLR, TKKLEYEVDA, LRVIYVLSST, VTEIVITYRH, SESTYKIKRK, STSIEIKYKH, IEITYVNHET, KKLVEQQYNH, NSSSQRQNSK, KTTTEQRYDH, NLQQQTNSET, IDRHNYTYTN, STSEHEIEVK, TVKVEYEYNN, HNETYIETVT, VNTFTYNYTV, SNYTYTQTTT, SQTVTYTYNL, FTYTYTQSTT, VNEVRVEYRG, STVRNRVVTK, SKTVEVRYNG, SEVENRITTT, VNNNNNNYNN, SSGTYNNEVR, STKNNNNYNN, NNVNNTESTT, VDRITKTHTN, STSEHEKTVT, HTTIKKEHNN, SRITKTKSVT, VLTVTVTVTV, VTVTVTDTVR, SRTVTVTVTV, VTVTTTDQVT, QVEVTVTYNV, STSTYKITQT, RKTVEVKYNV, VEVTYTITQT, VTTVSTDNIF, SSSTNTTTTI, SKNVSTTNNF, VSTDTNVSVT, VQTYEFKLRV, SELQLTYTET, SKEYEFTLNV, LTFKFRYKET, TTTVSVSATL, QDVSVTITEK, TKTVTVSATL, VTVSVTMTTT, IVTTKNTYKV, SESSYENTKR, SETSENEYNV, TTVKNKTKVT, IDRYQYEYSN, STYKYEREVR, SVTNRYEYSN, YTYQYERTVT, ERVKKNTQTQ, STHEHTREVT, SQEEKNTQNQ, KVTTHLVRTT, ITTRTSTRTL, STSTDIVTQQ, SVTRISTRTL, STQTDTVSQT, VQTITVTVRV, TESTVTVTRR, TETITVTVNV, ITVTVTDSVT, VEEVTVTYNT, STVEVRKKKI, TEKTEVRYNT, VEVTYKKIVT, VEKQVQKYTT, SNQTNDNTTT, KKSQNQDYNT, QTQKNATTRT, VLTNTNTTTN, STSTNTNTTT, KKTVTNTTNN, SSNTTTNTET, VVQVTAKVNV, SNAEVTVTTR, TNTVEVTVNV, VQTKVTVRDT, IERHEYTYRV, SESEYEKKEK, SQTEKAEYNV, YSHKYERSVT, EAEIVGVVTL, STSQVQVKVV, TVEIEGQVTL, IEGVGKVMVT, VHTVVVTVTV, STVQVKVEVT, SKTVKVQVNV, VTVTVEVKVT, ITEISISISI, SNIDINIKSE, KNKINININI, VEISISIEVT, VNTNTNTDTE, NTNTNTNKVI, KETNVNTDTE, NTNTNKNQRT, VDKHEHIIRI, ATINITNVQT, KQSKTVTISI, ITHIHHNTVT, IDTNTNNITI, SNSTNTKTTI, TQNNTVTINI, NTNTNTSRVT, VKRVTVIVRV, VEVKVEVRVV, TTAVEVEVNV, VTVIVEVSVT, VETVTVTYQV, SQSTYTYKKT, GEEETVTYNV, VTVTYTKKVT, KVTVTITTSL, STITIEITTT, TVKVEIETNL, VTITITTQIT, TLNVLVTLRN, SNISVEYKIT, SRSVEVELNN, VNVTVKYSET, TTTVTVTVRV, SEVTVTVTTT, KKTVVVTVNV, VTVTVTTTQT, VLEVTVTVTV, STVTVTFNLV, SKKVTVTVTV, VTVTVNFSVT, VESKSSTKVQ, YNSEKVSKEE, KRTKKSVKNQ, YSSTSSIKET, VVNLTLTYTV, FNLELTVVVT, KVNLTLTYNV, LNLTLTVSVT, VEVYTYTVTV, YTYTYTDTVV, SEKKEYTVNV, YTYTYSVSQT, QITVNVTVSL, STVTVTNNVT, KVNVTVTVNL, VTVTVNNLRT, INKVVVVVEL,VAVNVELTKT, KETKEVEVEL, VKVVVRQTVT, TETNINVYTN, NTNVVVVERT, KTKVKNVYTN, NTVVYIVDVT, VLKINVTYTQ, SNSTYTVEVV, SVKIIVTYTQ, ITVNVEVQVT, VREVTVSVDV, SNVSVSDKET, SREVSVSVDV, VKVSVIDKET, VDRVEVYYKN, SSYLYRHTRT, TTRVEVRYDN, YSVYYSIQTT, VLTVTVTVNV, VTVTVNHTET, RVSDTHNVNV, VTVTVSRTVT, VETTVYTYTH, SQSTVTYTRV, TSTTTYTYNH, TTTTYVYTRT, VENVIVTYTA, VTVKVEVTRK, SEEVEVEYQA, VNVYYKRHQT, VDTVTVTYTV, STTTVTYTQT, TKTVTVTYTV, VTVTYTHTTT, TVTVTVTVTS, STVTVTVTVT, TKTVTVTVNS, VTVTVTVTQT, VHVITRNDTR, SNSQDQQRTV, SVTIQRQDNR, SVINEQYTTT, KVNIDVKFKA, LSVEVEVTFN, SVTITVEFNA, INVKVEVSVT, VDKVTVTVTQ, STVSVEVTTE, RKKVKVEVTQ, VKVTVEVQET, VNTFTYEYTL, STFTVKNTQK, SNTTKYKYNL, FTYEYTNKET, TETDTKTVTV, HNYTVTKTTK, SKTDSKTVTV, DTYTKTVSET, VQMITVTYTV, STSTYTVTVT, RNTITVTYTV, ITVTYTLTVT, VNTTTVTYTD, STSKTTVTNV, TTTTEVTYTD, STVTVTVRMT, TLTYTYTHTI, STYTHTNTQI, SNTYIYTHNI, DSYTTTNSVT, VLQSRVLVEL, TRVSVEEKKV, NVLNEVEVEL, TRVLVKESVT, VDTLTLTYNL, STSTLSITKT, TTTTSVSYNL, LTLTLTNQVT, KVTVNTNYTN, SNISVEVTVQ, STTVTTEYNN, VTTNVTVSTT, KDKNKNTQTV, DSKDIDQEQT, TNKKKNDQTV, NKETNEQTIT, RVTVTVTVTT, STVTVTNVTT, TNEVTITVTT, VTVTVKNTTT, VRTFTSTNTT, STSTNTYKET, TKEYTSTNTT, FTSTNNERVT, ILRVTVSVTL, STVEVEVTQQ, STTVEVEVDL, VNVSVTHAQT, KSTNTNTNTN, NTNGNTNEQV, KSKNVTTNNN, NTQTNENQVT, VR IIIIIITY, STSAITISVS, TV IIIIIISY, TTTTITIKVT, IRKVKYEYSV, SNYEYEYEVK, SKRVEYEYNV, VKYEYVKEET, VTAVQTQLNL, SNVTVQIQQT, KTTQQTQLDL, VQVQVQQTTT, VDRVEYTYTL, SSTSYRTERR, SDRVLYRYNL, VTVTYESSRT, VVNITVSYSN, SNSTVTVTTK, STTITVTYSN, ITVSVTVSVT, VNERIIEDKD, SNRKKEENTR, TRDREEEDVD, REELKDETVT, VVTNTYTYRL, STSTYTHTVT, TERNTYTYDL, NTNTYTVQVT, QHTVTVTVSV, SSVQVQNTKT, SQTVQVQVNV, VTVTVTDKTT, TLTLVLSSSV, MSLSSSLTIT, TKTLSLSSNV, LTLSSTLSVT, VTTLTQVNVN, STSVQTQTVT, TKTLTNVNNN, STQVQTVKVT, VLTISISVTV, SEITVTVTLQ, RQSITTTVTV, ITISVSVSVT, TNNVSVSYKV, VEDSVSVNNK, TTTVVVSYNV, VNVSVQHNTT, INEVTVTVTA, LTVTVTVKNE, TKTKTVTVNA, VEVTVKNTVT, TLVVTVTYTN, SSSTVKVTLK, SVTVKVTYTN, VTVTVTVNVT, VNRFMFNANG, STFAIYIEQR, STKVSFAANG, FRFNAEHRET, VETYTGTSSI, YSYTSVETVR, TQTKVGVSVI, YTGTSSEQRT, VTKVEVTYTL, LTVSVEFETK, SEKVEVNYTL, VNVKVEFQST, ITRVRVTASI, YTVTIEVQRE, KQTVKVEAEI, VRVTIERTET, VVNVQVTVTV, STVVVVVQVR, SRTVVVVVNV, VNVTVQVHST, VSEKEIRVEV, STSEIEYTEV, AEEKEIEVEV, IEIRITYKRT, TLSININVDA, IGITVTVNTN, SVNINITVDA, ISINVNVSET, VQTNTNTYTV, NTSTIQNTQA, SQTNQITYTV, NTNVNTNQQT, SYTVTSTTKN, STVTTTYNAN, SRTVTYTTNN, VTVTSTYSET, KDTVIITVTD, SSVTVTINTT, KKTVNITVND, VTVTKINSTT, VNNVQYTYSV, STSTYTFQAT, TLTVEYTYNV, VSVQYKAQVT, VLTATFNYTV, SSSTYNYTVT, TRRVEFNYNV, ATFNYTTTET, VRVSVDTDND, SNSEVEINVK, TRTAEIEDVD, SVNTDKSTTT, VQIKETNTSL, SKSKTKVKVK, TTTKETKTNL, KETNTKVQIT, VTLVTVTYTI, STVTYTNNTR, SNRTTVTYNI, VTVTYTNHVT, KTIVNVDYSA, FNFSYTVTRV, STTTKITYNA, YNVDYTVSVT, TTSIQKQVSL, QQSSVQVTVT, SKTIQKQVNL, ISIQKTVQTT, IRRENSESRN, SNTRNEETRK, SRERRVESQN, SNTEVLESET, VLEVKVNYTN, SSVTVEVKVT, TKEVEVEYNN,VTVNVKVTVT, VVNVTVTVSL, SNSEVEVTVN, SVKVEVEVNL, VNVTVTVSVT, VLEVIVNVKL, SSSTVEVRTT, TKEVEVEVNL, VSVIVRVRVT, VREIEYTVRD, SESRVENKET, SNETVYEVND, IQYTYKTKET, VNKVEINKTD, STIRKEEEEV, RKKIEIEKND, VSIEEEEEVT, IWEDKKTKTK, DTQTDTEKRT, TRTDEATDTK, DTVTEKTQVT, KVTVKVKITI, STSEIEVTVT, SVKVEIEITI, VTVKVVVQTT, VDNVNVNVTV, VTVQYSYTLT, SKQVSVSVTV, VAVNYNYKET, VERRTHTDTV, RTNTTTNTRK, TVSTVITDDV, RRSTHSLTET, VNSVTYTYSL, STESVSVTVA, KTTVSISYNL, VSVTYTVSVT, VSTQSISVTI, SSSNINNTEK, SKTQNINVNI, ITISITNKST, SEEHEHEVNV, SESEEEEEVK, SREHEHEVNV, HKHEEEEKVT, VVNVDVTISL, STTSVTVTVT, KVKVTVRINL, VDVKVEVTQT, ASSVLVSYRN, SNVTLSVYTE, KRLVKVSYTN, VTVTVTVKTT, VNKVTVQYNL, STSEVENTTA, SQTSTVEYNL, VKQQVTNSTT, INKVTITVNL, GNVTVTQQQK, SRTVIITVNL, VTITVQQQVT, VNLVINIYNV, SQSTVVNVKT, KTLVVNVYNV, VQVINVVAKT, QYSVSYNYSV, ASVSVTVNVT, KVTVTVTYTV, VSVSYSVTVT, VVTVTVTLTN, SSLTVTVVVT, KRTVTLTLNN, VTVTVNVTVT, TAKNNFKNTN, SSSTNEVTVT, SVKVEFENNN, NTFKFSVSVT, VDSNVNKASV, SNSEAENTTK, SKEIENEAQV, NSNTNKNAVT, VNIVTVTIKL, SEVSISVKVV, RVTVIVSITL, VTVTVKVTVT, VRYQRLRYTQ, STSEYELEET, SRSQQLEYNQ, QTHRYELRTT, VANVKSEYKV, SESEVEVVVR, SEEVKSEYKV, VLVEYQVSVT, TLSITVTNSV, SNSTINVTVT, KVSINVNNNV, ISITVAVTTT, VNSLSINYSN, SSSNYNVVKT, TDTLNINYNN, LQINYVVKVT, RLEVSVSVNV, KSVSVSKTVV, TVRVEVSVNV, VEVSVSKQVT, VDRSEITYNH, STSRYEIERQ, KTRSEIEYNH, SRITIEKTET, VREVKVTYKN, SESTYTVKTT, STERIVTYNN, VTVTYKVKET, RREVKVKITV, VSVEIEVVSI, TNEVEVEITV, VTVKVKVQVT, TITTTYTYRD, SESTYTQQQQ, SNTTTYTYSD, TTYTYTQKQT, VDTNTNTYSY, SSITNTNIQR, SNTSTQTYNY, NTNTYTNSVT, KRTVITTYNN, STKTVTDQNT, SEKVETTYNN, VTVTVLDQQT, IYTAQTTDTV, STSTDTHQIT, STTKIITDTV, ATNKDQRSET, LKVNKIQIQL, SVSEIEQQVK, SVKNEIEIQL, NVIQIEQKIT, KNKKEVTGRG, ATSTVTVERT, SERKTVTGNG, LTVTVEVQQT, RVVVEVELTV, VNVSLRVETR, TRAVRVRLTV, VTVELEVQET, TTTQTQTVTI, STQTVTVTTT, SQKQTQTVNI, QTQTVTVSQT, KVVVKVVYTI, SSVSYDVTIT, KVTVDVDYTI, VTVVYTVSVT, KLRLELELTA, LTLNLEVEVT, TKKLKLELTA, LRVELEVQVT, INTVTVVVTV, YTNVVVVTVI, TVTVVVVVNV, VTNVVTVTVT, IDESEYKVSV, SNYTVSYTAT, TKESSYSVNV, SEYKYTLKWT, VVKVEVEVTD, SSSKVTHEIR, SEKITVTVND, VEVEVETQVT, VTQTTVTESQ, SHSTISVLTT, KTTTTVSENQ, TTTTETVTTT, VNTVTYTVAV, STSTVSHTQV, RTTVSVTVNV, VTVTYTHTET, VNRSVNKNSN, SSSTVEVVTK, TNSVEVENNN, SRSVNVVKTT, IQIVNTTYNA, LTINDNVERK, STKVITNYSA, LNVSYKDEVT, VVTVTTTYTV, SNSTIVTTVK, SVSVVTVYTV, LTTTTVTSVT, VESVTVTVTI, SNVTVTVKVT, TKEVTVTVTI, VSVTVLVKTT, VVTITVTYTL, SKITVTVTNV, KKTITVTYNL, ITVTKTVSVT, VLTNSNSNTN, STNININTVK, TTSQININNN, NTNSNTVRTT, IENNMYRYTD, SNYSNQQTVT, TNDNQYQYND, NNYRYTKRVT, VQSNTETKTH, NTSQKSVTKT, TETNSVSKSH, TTITETVKTT, VTVNTIIVTD, SNIRVEIRVS, SVENEIEVTD, INIIHREYVT, ISNNTNTKEV, STNSNSNTTT, SNTNSNSKEV, NNNTNTNKVT, INSNTSTTSL, NSNTTSSTRK, TKSNSSTTSL, NTSTSSKRVT, VTTVQVTYNI, SSVTYTEEER, TKTVTVTYNI, VTVTYQEQVT, TLTTTVTVTN, STNTVTVTVK, AVTKTVTVTN, TTVTVNVSMT, INQNENKYTL, STYEYKNKLT, TKQNINKYQL, NQYENENKIT, IIKIKITITV,STSEIEIIVK, SVKIKIEITV, IKITISIEVT, VTTSLNNYNN, SDSSITNNTK, STTSTNTYNN, STNQINNSVT, QLNNNYNYNL, SNSNYNFSLK, SKTNNFNYNL, NTSNYSWTNT, VVSLTLNLEV, SSFTYTITVT, RVTLSLTLSV, YTLNLTVSVT, VYKIEIKISV, VTIEIKIEKQ, STKIKIKINV, ITIKIEASVT, VRTWEVALTN, SSVKFKYIDR, SRTWIVKLNN, STVAFKYSLT, TVTTTVSVSN, SNTTVNYSVT, RKTTNVNVNN, TTVSVTYITT, VVENLTQVSV, FSNTVNSKQT, RQENNHNASV, NSNQTKSLQT, TETSTSIYTA, STSITKKETT, RKNTKSKYNA, STSIYTKKVT, VDTNTNTNTQ, STNTNTTTTK, TNTKTNTNTQ, NTNTNTITVT, VDTKTVTVNL, STSTETVTTK, STTKTVTVNL, KTKTVTRKET, VRTTTVTYSV, STSTYSDTET, SKTTSVSYTV, TTVTYTDTTT, KRSVTTNYQI, SLSTVEVEQT, SRKVTVEYNI, VSVKVTEQET, VNRVVYNYNL, VSVEVENINR, SQRVEIEYNL, VEYRYVNEQT, VETVSVSVNI, SSVKVEDTVT, KKEVKVEVNI, VNVSVTSLTT, VNHNENTNTA, STSVNNNTNV, KNDNVNNNTA, NTNTNRNSVT, NLSSTNTNTN, STSNNNVDVT, TTTDEVNNTN, STTTNKVTIT, IVTNINTNTT, STNTNLNTVI, RRTNLNLNNT, NTNTNLLTVT, VLEVEVTLSQ, STSSVKVKVT, QKEVEVKLNQ, VEVKVKYTRT, VWKNENEIKL, SESKNKNEKK, TVTEKNKINL, TKNENEQKVT, VTRNEQEYTN, STIEREEEEV, TIKNKQEYTN, ITQEQEERKT, VATVTVTYTV, STSTVEVTTT, TTRVEVEYTV, VTVTYTVKTT, KNNNYNGATI, LTNNNNNNTT, TNTNNNNNNI, NNNYFTNTQT, RKTIITRYSL, SNIEVEYDEQ, SRKIRTEYNL, ISIRYDYQTT, VDSYQYQYEL, STYSYTDTKV, TVKEEYTYTL, YTYQYDRQVT, VLYVSVTATL, SSVTVTYEET, RKKVTVTATL, VSVTVSYEST, KLSWDYTYHV, SEWDVDYLQQ, TKTWDIDYAI, WSWNYTYTQT, VDIIVHTDRV, STSIDVQRIS, SERRVHIDNV, SSHTHESHRT, VNTVTVTLQN, STDTVTNTQT, TNTVTVTLTN, VTVTVTNQVT, EARVQVEYTL, SSVKVENLTK, TRTNEVEYNL, VNVKYQNTVT, TEKTKNVNTK, SNSTNTHETT, TKTTTNTNNK, TKEVNKVETT, VDVRTNTNSV, STSTNKDNVT, SNSRKNTNNV, SNNTNENRVT, VVTVSVTVTL, STSSVSVTVT, SVTVSVSVSL, VTSTVTVTTT, TLTVTISYTT, STSIYEKTIK, SVTVKGEYTT, VTKSITKSVT, IKRVEVKVQV, VNVEVEVEET, KKQVEVEVNV, VTVRVEVLET, VNEYEYHLTV, STSRYTNRNV, SKSYTNTLTV, YEYHYRNSVT, VISVKVIDRD, STSEVEVKTV, TVKVEVEDTD, VSVIVKVEVT, VQTRTSINKH, SESTNTVEVK, TTKNTKTNAH, RTSINSVEVT, TLETTVSESH, SSVKENRTEV, STTTEVNEVH, REVSEKRSVT, IQTTTVTLTL, STSVVVQTQR, TTTTVVVLNL, TTVTVTQKRT, VVETSLTEQD, SESNETQKER, SVETSQTEQD, TTLTLNQRRT, VLSVTHTNTN, STSTITYTTR, SRTVTHTNNN, VSHTNTYSVT, VLTVIVTVSV, VTSTVEVKVT, SVTVEVEVSV, VAVTVKYSVT, TLYVVVVVTV, NTSTVVVTVT, KVTRVVVVTV, VTAVVTATRT, VDTTLVTVNN, STSTTTHTTS, TKTNEVTVNN, TTVTVKVTET, VRTETFTARL, SSYTATFSQT, SRTETFTASL, ETFTFTFSVT, TVTLILTVTV, STSSISVTVT, KVTLVLSITV, LTLTVTVTVT, VVSVSISVSV, VSSTVTISVK, TTTVTITVNV, VSISISIKQT, TVTVTVSATL, STSTATVTIS, SRTVEVTARL, VTVSARVQET, SVNNNNNNNL, NTSNNNNSTQ, TERNENNNNL, NNNNNTIQRT, VSKRSYTYTN, STDSYSIEQK, SKTTSISYNN, RDATYEETQT, TTEVTVTVTL, STVTVTIKNV, SKTVTVTVTL, VEVTVTDSVT, ILKVVVDYRN, SNVSVSVKVT, SVEVSVSYDN, VKVDVTLKLT, ITKTKVVYNN, SNSDYDVEVR, TVKTTVDYNN, TKVVVEVEVT, TRTVTVTYTA, STSTVVHTER, SETTVVTYNA, VSVTYTHQVT, IDEISYTYKI, STITYNYKKI, SKNITVNYTI, IEITYSKHET, VRMTMTTVQV, STSSVSTVVF, SVTTSTSVSV, TMTTVVVSVT, VASTTTTYTL, SKSTNTITLT, TVETETTYTL, TSTTYKLTTT, VTTYTHTTRT, RETYTHTTST, YTHTYTNTRT,VDTNRNTNTN, STNENENTKT, STKNENENTN, NTNVNTRAVT, KTTNTNTNTT, NTTTNTQVLQ, SKNSTNTNTT, NTNTNNQQTT, VTVEVTVESG, RGTVRVTVRV, ITITITITVT, TVRLRITITI, ETVTKTKTKT, VVTETETKTK, TVEVTVTVTV, VTVTVVVTVT,

[0501]

[0502] IVIVIVIVIT, NATIVIVIVI, NSTLTLTVTV, KTYTKTKTKT, IELKTKTKTK, TATVTVTVTV, DTDTDTDTTT, TTTDTDTDTD, VVVVVVVVVS, TVTVTVVVVV, ATATATATVT, TVTYTATATA, VTVTVTVTVD, KISVSVTVTS, ITITITITIT, NVTITITITI, ATATLTATAT, TV I I I I I ATA, VTVTVTVTLT, TSTVTVTLTV, LTLTLTLTLT, TATLTLTLTL, NTNTNTNTTT, KKTNTNTNTN, LVLVLVLVLT, TVTLVLVLVL, TVTVTLTVTV, VVEITITITI, ATATATATAT, NLSVTATATA, AEAEAEAEAK, VAEAEAEAEA, TVTITITITI, STNTNTNTIT, TKLVTNTNTN, TLTVTVTVTV, VVVVVVVVIK, SLEVEVVVVV, TVRVTVTVTV, 11 IIIIII LT, TATTTTTTT I I, FTFTFTFTFE, QLSFTFTFTF, DVQIVITITI, VVVVVVVTVT, TVKVVVVVVV, IVHVHVHVHT, KIEIVIVHVH,

[0503]

[0504] SISIHISIST, VKQSISISIS, TRTDTDTDTT, TVKITITITI, YTVTVTVTVK, IVEVTVTVTV, TITITVTVTV, YTYTYTYTVT, TVTYTYTLTY, TIKVTITITI, ITITFTITLT, TSTITITITI, TVTVTITITI, QQQQQQQQQT, TVTDQQQQQQ, SLTVTATATA, ITITITVTVT, VEVEVEKTKK, EEEEVVEVEV, VEVEVEVEVE, KAEVVVEVEV, TITITITITI, TGSTITITIT, LDLDLDLDLD, EFDADLDLDL, ITITITITIE, KITITITITI, TISITLTLTL, KIEITITITI, HTHTHTHTHT, TTRTTATKTH, SVTITITITI, TITITITIVI, IVVVVTITIT, DTDTDTDTDT, VDVDTDTDTD, TIKVTVTVTV, TIIITITITI, NVNVNVNVNT, TGVNVNVNVN, TTTITITITI, ITITITISIS, NVKITITLTI, LVLVLVLVLK, TVEVVLVLVL, LKLKLKLTLT, TATLKLKLKL, VEVEVEVTVT, TKKKEVEVTV, NTNTNTNTNT, KTTNTNTNTN, VQVQVQVQVT, QARVQVQVQV, ITITIVITIR, TAEVTVTITI, TTLTLTQTQT, TSS IIIIII Q, KIELTVTVTV, TVTFTLTLTL, IVIVIVIKIH, KVEIRIVIVI, VTATATATAT, KLDATATATA, LTLTLTLTVT, TVQLTLTLTL, VVVVVVVVVK, TVTVVVVVVV, TAVVTVTVTV, VTLTVVVTVT, TETVVVVVTV, KKSNTNTNTN, FEFEFEFEVT, TVKFKFEFEF, TIKIKITITI, KIEITIIIII, TAELTLTLTL, IKIKIKIEIE, EVKIKIKIKI, VEVEVEVVVT, EARVEVEVEV, VTVTVTVTVV, ELKVTVTVTV, ITITITITIQ, KVSVTITITI, KVSITITITI, TKVNVNVNTN, TITVTVTVTV, TTDTQTQTQT, TN I I I I I QTQ, VTVTVTVTVK, TVVVTVTVTV, ISISISIKIK, TLELSISISI, LTQTQTQTQT, KLTQTQTQTQ, TVEVEVEVTV, TVNITITITI, LTVTVTLTVT, EGKLTLTVTV, QIQIQVQIQT, TQKQIQIQIQ, TLTITITVTV, LVLTVTVTVT, TLKIKLTNTN, GTGTGTGTGE, LGRVTGTGTG, YVYVYVYVYT, KKENVYVYVY, SVSVSVSVVV, SVKSVSVSVS, TTEKTVTVTV, TVTHTHTHTH, ITITITITIK, TLTVTITITI, FTFTFTFTFK, TITFTFTFTF, NTNTNTHTHT, TNTNTNTNTH, AVAVAVAVAT, KVSISIVAVA, ITFTFTFTFT, TVTVTITFTF, TTTTTTTT I I, TAAATATTTT, KIEIVVVVVV, ITVTVTVTAI, KSEVTVTVTV, VTVTVTVTIT, TIEVTVTVTI, NTTVTVTVTV, LTLTITITIK, ITITITLTIT, TVKVVIVIVI, VTVTVIVTVT, TKTVTVTVTV, EVEIVVVVVV, MVMVMVIVIT, KAQMVMVMVM, YTYTYVLTLK, DIEEEYTYTL, TVSVTVTVTV, TLRATATVTV, VTVTLTVTVT, TLSVTVTITV, TVKVTITITI, DVNVNVNVVT, TKTKTNVNVN, ITITIIITIT, NINVTITITI, VIVIVIVIVK, TVEVIVIVIV, LTLTLTLTIT, KVELTLTLTL, AIAIAIAIAT, TVELIAIAIA, TVKVTVTVTV, KLTYTYTYTY, HI HI HI HI HR, QEEAIHIHIH,

[0505]

[0506] ITVTITITIT, KVEITITITI, DVKVTVTVTV, ETETEKEKEK, TETETETEKE, LTLTLTLKLK, VEEQELELTL, TVKITITVTI, TVTVTVTVTT, TVTTVTVTVT, VVIVIVIVIT, KITVVVVIVI,ATATATATRE, TVQETETATA, ITVTVTVTVT, TVTITITVTV, TVTITLTLTL, KVTVTVTVTV, TVTVTITVTV, TLEATATATA, VVEVTVTVTV, STSTSISTVT, TATSTSISTS, HIYTYTYTTT, TRTHTHTHTH, TLVVTATATA, TININITITI, LTITITITIT, TTKITITITI, YKYKYKYEQK, EKKYKYKYKQ,

[0507]

[0508] TITLTLVVTV, NTNTNTNTST, INTNTNTNTN, TIKVKVVVTV, QTVTVTVTVT, KKTQTQTVTV, YTITITITIT, TIEVTITITI, VAEIVITITI, KATAIAIEIA, NTNENENTNT, DVKNKNENTN, FTFTFTFTFT, TVSVTFTFTF, TETNTNVNVN, SNSNYNNETT, SREHTHTHTH, VVVVVVVVVT, TVVVVVVVVV, VTATVTVTVT, TISVTVTVTV, ATATATATIT, TVTATATATA, NVKSTITITI, NTNTNTNTNN, ETWNTNTNTN, VTVTITITVK, TVEVEVTVTV, ITITIMITIT, IGKIMIMITI, TVNFTFTFTF, STSTSTSTVT, TVTSTSTSTS, VVRVVVVVTV, TVTFTFTFTF, TVSVTITITI, VTITITITIT, TVAVTVTVTI, LTLTLTLTAT, TAILTLTLTL, VTNQNTNTNT, ILEITITNTN, GTGTG I I I I I, TAQGTGTGTT, DVDVDVDVDT, TDVDVDVDVD, KVQITITITI, ATLTLTLTVT, TATATATLTL, VTVTITVTVT, AVKVTVIVTV, STTNTNTNTN, KKEIEHTHTH, VIVIVEVVVT, TEKVEVEVIV, TATLTLTVTV, VTVTVTLTVT, TVKVTVTTTT, EVKSTSTVTS, TVTVTVTLTV, VTITITITIK, IEIEIEIMIK, TVEVEVEIEI, YTNTNTNTNT, VQKKTKTNTN, QATVTVTVTV, TLVLTITITI, KVKVKVTVTV, TLRIIIIII I, DVEVTVTVTI, TLTLTLTLTL, IKIKIKITIT, VKEVKIKIKI, LTLTLTITIT, IVELTLTLTI, ITITIVITIT, NINITITITI, KVTITITITI, LILILILIVT, TAKLILILIL, TIEVTVTVTV, QTQTQTQTQT, TNTQTQTQTQ, TKQKTHTHTH, IEIEIEIEIE, EVKVEIEIEI, TKKNTNTNTN,

[0509]

[0510] TTRVTVTATA, GTGTGTGTGT, NIRVTVTGTG, YTYTYTYTYT, TRKYTYTYTY, TIKITITITI, KVSVTVTVTI, TIKLTFTFTF, TAQLTLTLTL, LTLTLTYTYT, TITLTLTLTY, KIKIKIKTKT, TATAIAIKIK, TKEYVYTYTY, HVHTHTHTVT, EIKKKHEHTH, EVKVVLVVTV, TAKITITITI, KKEKTDTDTD, TATVTITITI, VTVTLTVTVK, KVDVTVTVTV, TAQATLTLTI, ITTNTNTNTN, RLELVITITI, TVKVTYTYTY, TALLTVTVTV, TVTNTNTNTN, FTFTFTFTVT, RVEFTFTFTV, ETETETETVT, TNTNTETETE, TVSNTNTNTN, ATVTVTVTVK, TVSATATVTV, SIKVTVTVTV, VATHTHTHTH, AVATATATVT, TAEVKVTVTV, VTVVVTVTVT, KVEVTVTVTV, ININNENKTT, TLTATATATA, DVRAVAVAVA, NLNLNVNLVT, TVTVLNLNLN, NTNTNTNTVT, TTKNKNTNTN, IEIQIQIQIK, VITIQIQIQI, YTYTGVGTGT, TVTVTYTYTY, TLQITITITI, TVTLTLTLTL, VVVVVVVEVT, EAKVVVVVVV,

[0511] TTTTTTTT I I, TVTTN I I I I I, VTVVVVVVVT, TVEVVVVVVV, TVTATAAATV, LVVVVVVVVT, TVETVVVVVV, YTYTYTYTYS, TLTLTLTYTY, TVKVTVTITI, TVTVTVTVTL, TATYTITITI, NVVVTVTVTV, VTETETETET, TTTVTVTETE, DIEATATATA, TLEIKITITI, VVRVTVTVTV, KTKTKTSTKT, TVTKTKTKTK, FVYVYVYVYT, EIRFVFVYVY, NTNTNTNTIT, VKKKTNTNTN, SKTINTNYNA, TISSSSTITI, TTTTTTTT KT, TTTTV I I I I I, ITATATATAT, KTAETAVATA, TKQKTQTQTQ, YTYTYTVTVT, TYTYTYTYTV, VTLTLTLTFT, SVRVTVTLTL, VVGVVVTVTV, ITKTKTKTKT, VVTETKTKTK, ETKITITITI, FTFVFVFTFT, TVKVKFVFTF, TGTLTLTLTL, ISISISISIS, TLLVSISISI, I I I I I VTSTS, TVTTTTTTTT, DGKVTVTVTV, LTITITITVT, TKKITITITI, TQTVTVTVTV, TLTVVLVLVL, LTVTVTVTVT, TVVATATATA, QTQTQTQSQS, DKKHTQTQTQ, TKEVIVIVIV, NIKITITVTV, VKVKVKVTVT, TVEVKVKVKV, ATSTSTSTVT, IVTATATSTS, IEITITITVT, IIKITITITI, NLNLNLNLNT, KNNNQNLNLN, VIVIVIVIVT, TIQVIVIVIV, IVIVIVIVIK, EVEIEIVIVI, DIDIDIDIDT, KRERSDIDID, VSVSVSVSVS, TLTVSVSVSV, TVTVTVTVTK,EEQTVTVTVT, IVIVIVITIT, TAKIVIVIVI, QIQIQIQIVE, NKKQIQIQIQ, ITITIVITIK, EIEITIVITI, NININININT, TKTNINININ, ETETYTGTGT, VREETETETG, TVEITITITI, NITVTVTVTV, VTVTVTVEVL, VVKVKVKVEV, TLTATVTVTV, TLTLTVTATV, NLTVTATATA, ITTTNENSVS, TVKVKTTTST, VTNTVTVTVT, TVRVRVTNTN, ITITITITKT, TLTITITITI, VTVTRTRTRT, TEKRTRTRTR, TNTNTNTNTN, TTTDTVTVTV, ITQTQTQTQN, TNTSTSTQTQ, TKMSTNTNTN, TVQVQVTVTV, VTVTVTVTFT, TNTLTVTVTV, VVKITITITV, AIAIAIAIAK, VVEAIAIAIA, TTTNTNTNTN, VTVTVTATAT, TLTVTVTATA, QTTVTVTITV, TVTVTVSVTV, VTLTVTVTVT, TLELKLTLTL, VTVSVSVSVQ, KETVTVSVSV, LILILILILT, AATLILILIL, RVEYTYTYTV, ITITITITVK, EIEIEITITI, EVEVEVTVTV, ITITYTYTKK, TKEITITITY, ITITIKITVK, KVEIEIKITI, TVLVVATATA, NTNTNTNKNT, TKVTDNDNDN, ATGTGTGTGT, TKTATATGTG, TVTVTFTFTF, TVSITITITI, IVVVVVVTVT, KKEEVVVVVV, NTNTNTNTKT, SVQITITITI, KRSDTDTVTV, VTVTATATAS, VANVTVTVTA, AVEVTVTVTV, TVKVTVTNTN, VTNTNTNTNT, LTQSTNENTN, TPTGTGTGTM, VTITITVTVN, VTQVTVTVTV, NTNINININI, KKQNINININ, TVKLTARATA, TAEVTYTYTY, FTFTFTFTIT, KIEFVFTFTF, TAKATATATA, TVELILILIV, VTVTVTVTYT, VKKEKQTQTQ, AVAVAVAVAK, NVELVAVAVA, IVVVVVVVVV, TVFVVVVVVV, TIGVVVVVVV, EVTVTVTVTV, IVTVTVTITI, EVKITITITI, VTVTVTVTVE, TAKVTVTVTV, IVIVIDNVNK, ETEIVIVIVN, IQETITITIT, VAEFKFTFTF, IVTLTLTITI, TVEVTVTITI, TALVTVTVTV, IEIEIKIEVK, TITIAIAIEI, TVTAVVVVVV, TVKITITLTI, TITIIIIII I, TISITITITI, TSTVTVTITI, LVLVLVLTVT, VLTLVLVLVL, NVSITITITV, IEIVIVIVIS, TVSAVAVIVI, KTKTKTKTKT, TASVTVTVTK, VTTTVTVTVT, KATVVVTVTV, EKEVTVTVTV, TIKLTLTLTL, STSTSVSTVT, TVTSTSVSTS, NTNTNTNTNK, TETDTDTDTD, HTHTHTHTIK, TITQTHTHTH, NTNTNTNTLT, TAVNVNTNTN, QIQIQIQTQT, EKKQIQIQIQ, VIVIVIVTVT, KVEVIVIVIV, YAKITITVTV, TAKVVVVVVV, KNKIETNYTA, VTVTVTITVT, KATVTVTVTV, VRTNTNTNTN, FTITITITIT, TATLVLVLVL, ITITIVITIE, TIKKVKTITI, TARSTATATA, HVHTHTHTHT, VRTETHTHTH, TFEIEITVTV, VTVTNTNTNT, TKSVTVTVTN, VTVTVTVTAT, NVTITITVTV, VTVTITITVT, TVKVTVIVTV, RTRTRTRTVT, EKKDTRTRTR, IEIEIEITIT, TLKIEIEIEI, NININVNINT, TKKNINININ, TQNVTLTLTL, VVVVVVVVAT, EERVVVVVVA, RTATATATAT, QVKVTVTVTV, LELELELESE, TRKIKLELEL, QTQTQTQTTT, TKTETQTQTQ, DTRTDTHTDT, TRVDTDTDTD, VTGTVTVTIT, NTNVNKNSVT, TVTV IIIIII, DKKNTNTNTN, HIHIHIHIVK, VREEIHIHIH, LTATATATAT, GTGTGTGTVT, TVVGVGTGTG, ETETETETET, VREVTVTETE, YIS IIIIII I, INITVVITIT, TRTITIVITI, ITITIHITIT, KVSLSLTLTL, SVTVTITITI, TVTGTGTGTG, TVSVSVEVEV, TITVTVTITV,

[0512]

[0513] NAVVVVVVTV, LTLTLTLTLS, SISVTVTLTL, TLEVTVTVTV, QVKITITITI, TVTVVIVITI, ATATATATVK, TATATATATV, TITVTVIVIV, AIAILILILT, TVTATAIAIL, VTVTVCVTVT, YELELRLRLT, GLTLTLTLRL, KVEVIITITI, IIIIII HTHT, SESESETHTH, VQVQVQVTVT, TVNVQVQVQV, ITITISITIT, TIEITITITI, VVTLTLTLTL, TTNVTVTVTV, MVMVMVMVMT, TVEMVMVMVM, TKRVTVTNTN, DTDTDTDTVT, TVRVVVVDTD, TVTKTVTVTV, VTVTVTVVVK, TTTTTTTT VT, TVKIIIIII I, VVVVVVVVVE, TVRVVVVVVV, STSTSTSTST, TISSTSTSTS, TKENTNTNTN, TKTATNTNTN, QVTITITITI, STSTSTSVAE, TVRSTSTSTS, NVKITITITI, LTLTLTLTLK, VVELTLTLTL, TETVTVTVTV, ITVTVEVTVT, I I I I I NTNTN, LTGTGTGTGT,KLTITGTGTG, NVNVNVNVNK, DKSNVNVNVN, IVIVIVIVIV, VSLVVIVIVI, TIEVNVTVTV, VAQFTFTFTF, NLKATATATK, KKTVTVTNTN, RIRIRIRIHT, TDKRIRIRIH, IVVVVVVVVT, TVEAVIVVVV, IEIEIEIKIK, TVEIEIEIEI, AVQVTVTVTV, ITITIEITIT, ELKIKIEIEI, AAAAAAAAVT, TSTATAAAAA, TATVTLTLTL, TQTYTYTYTY, VIKITITITI, TLGLTLTLTL, LTLTLTLTVK, TVELTLILTL, IEIEITITVT, TVKVEVEVEV, EETETKTKTK, TGKVTVTVTV, SVSVSVSVTV, TVRIVSVSVS, QVQVQVQVQT, TTTQVQVQVQ, DTDTDTDTKT, TVKTK I I I I I, TITVTVVVTV, KINVTITITF, TVTLTLVLVL, TQENTNTNTN, KTETETETQT, RTERRERETE, HQHQHQHQTQ, TRVHQHQHQH, TVEVTITITI, LVRVTVTVTV, HSHTHTHTCT, TVTVTHTHTC, VVIVIVIVIK, ELSVWVIVI, TEEATDTDTD, DVNVNVNVTV, IVKIKIKVIV, SVIVIVITVT, TAELVIVIVI, TFEVTFTFTF, TKTDTDTDTD, VTTTTTTTT I, TVTVTVTTTT, GVGVGVGVVT, TVEVVGVGVG, TIEVTVTITI, LELELELEVT, VVKLKLKLEL, TVTITIEIEI, ATATATATAV, ITEEEATATA, YTFTFTFTFT, DSTFTFTFTF, TVKIKITATI, KTKTKTKTKK, TEEKEKTKTK, QVKVKVTVTV, TVQVTITITI, VTITVTVTVT, TFEITITITI, ATATAIATAT, AVKDTAIATA, TKTNTNTNTN, KVTVTITITI, ITITITITFT, TYLGTITITI, TAQVTVTVTV, SNTNENENSV, AEAEAVAKAK, TAEAEAEAKA, KLTVTITITI, TATITIQITI, ITLTLTLTLT, TLNLNLTLTL, ITNTNTNTIT, TKTITITITI, TVKITVTVTV, ATITITITVS, TTTVTITITI, EVKVKVKVKT, EERETEVKVK, EIEIEVTVTV, LELELELELT, TATAELELEL, YTATATATVK, TVELTATATA, VKEIEIEITI, NISVTVTVTV, HQHTHTHTHT, TLSHSHTHTH, TVTITITFTI, LELELELELR, VLEVEVELEL, NININININA, INLNINININ, ITITLTITIT, VDTNKNTNSV, TVKATITITI, VEVTVTVTVT, TITITITVTV, NNTNTNTSTS, KTKTKTKTVT, TKTETKTKTK, ITVTTTVTVK, EIEIEIEIEI, KIQEIEIEIE, SVSVSVSVSV, TGTVTVVSVS, TVTATFTFTF, TKVNVNTNTN, VTVTITVTLT, TATVTVTITV, VTVTVVVEVD, VARVRVTVTV, ITIIITITIT, YTYTVTVVVA, TVAAIAIDTV, VIVIVIIIIT, TGKVIVIVII, AVAVAVATAT, TLEVVVVAVA, VTVTVTVTVS, TVKVTATATA, TVRVRVTVTV, LTLTLTLTLQ, TVTVTLTLTL, EVEITITITI, TVTDTDTDTD, QKTYTYTVTV,

[0514]

[0515] LSLSLSLTLT, TATLTLSLSL, NISISISISI, QSSSISISIS, LTLTLTNTNS, TSKLTLTLTN, RAQLQLQLTL, TLKVTVTVTV, TVTLTVTVTV, KTVTVTVTVT, FTFTLTLTLT, TATFTFTFTF, ATATLTATIA, IVVTITITIT, VVTITITITI, EKKNTNTNTN, LTLTLVLTLT, TLRLTLTLTL, AVAVAVAVAR, TVEVVAVAVA, TIRVTITITI, VVTFTFTFTF, KDHVHVHTHT, TTVKVKVHVH, VTLTLTLTIT, TVTVTVTLTL, LVLVLVLVVT, EIEINITITI, TVKVTKTKTK, KTKTDTKTKT, KKSKTKTKTK, KTKTKTETET, KKIKTKTKTK, NIKITITITI, VIKVTVTVTV, IVIVIVIVVT, EVEIVIVIVI, ELRVTITITI, VVDVVVVVAK, KLEVVVVVVV, TVVVTVTATA, TLTLTATATA, STNTNENTTK, NVTLTLTLTL, KLEIEITITI, VVVVVVVVVR, TATGTGTGTG, GVSVTVTVTV, TETKTKTKTK, NVNVNVNTNT, TKENVNVNVN, KVKVTVTVTV, VVEVVVVVVV, TAQIVIVIVI, TVQVTVTVTV, NVTLTVTVTV, VVIVIVIEIT, EIRVRVVIVI, TVELTLTLTL, EETKTKTKTK, SISISISIST, TSSSISISIS, TLVATITITI, ITITITITVS, FKFKFQFTVT, DVEVKFKFKF, TTETTNINTN, ATVTVTVTVT, TNTITITTTT, YTITITITVT, IMITITITIK, TATTTITITI, MTMTMTMKIK, TIEMTMTMTM, VTVTYTVTVT, NVTVTVTVTV, TIRITITITI, FTLTLTLTIT, DVTLTLTLTL, VTVTFTVTVT, TITVTVTLTV, ERRYTYTYTY, NLNITITITI, VTVTVVVVVV, FIFIFIFIFT, TGEFIFIFIF, VVVVVEVVVS, ILSITIIITI, TVKVIVIVIV, AVAVAVATVT, TTQEVAVAVA, VVTATVTLTV, TITVVIVIVI, NTNTNTITIT,TAKNTNTNTN, TTTVTVTVTV, EVKVEVTVTV, VTVTVVVTVK, EVEVEVVVTV, LTGTGTGTVT, EVEAVATGTG, TLSVSVVVVV, TNKITITITI, ITITITISLS, PLELTLTLTI, KKKYTYTYTY, TVKIEITITI, EIKITITITI, KEKEKVKKIK, TKEKEKVKKI, TLTYTYTYTY, TYTLTLTLTV, EVKVVVVVVV, TATLTLTLTV, NSNTNTNTNT, ETETETETKT, KETETETETE, VQVTVTVTVT, KITVTVVVTV, VEVEVEVKVK, EIEVEVEVEV, TIKG IIIIII, NTQVTSTSTS, KVEIEITITI, KTKTETETEE, TKKKTKTKTK, VTVTITITIT, TITVTVTVTI, VTVTNTVTVT, TKTNTNTVTV, ITETETETET, VKEIVITETE, LTVTVTVTVM, TVLVTVTVTV, TITATGTGTG, IDIDIDIDIT, TATIDIDIDI, TTKTVTVTTT, TTTTTTTT I E, TVRTR I I I I I, RLEIEIEITI, SVKLVLVLVL,

[0516] VIVIVIVIVS, TKTK IIIIII, TLTLTLILTL, TVEAEATATA, VVVTVVVVVV, AVVVVVVVVV, TGTATATATA, TVTETATATA, TLTVTLTLTL, TTKNTNTNTN, KVTLTLTITI, KITVTITITI, VTVTLTLTVT, TTEAKAKATA, LTLTLTVTVT, TLKLTLTLTL, VSVSVSVSVT, TTTVSVSVSV, TTKIVIVIVI, DVKVKVTVTV, VTVTVTVTIK, TVEVEVTVTI, AELTLTLTIT, TVTATATLTL, EITITITITI, TVKLTIVITI, ITNTHTHTHT, TETNTNTNTH, LTLTATATAT, NISITITVTV, KTKTKTKTIT, NKNKTKTKTK, KKEKTYTYTY, HTKTKTKTKT, THKDTDTKTK, ATVTLTVTVT, VVQVQVTVTV, EVRLTLTLTL, MTMTMTMTVT, KVELTLTMTM, ITITITITNT, TIKITITITN, VATVTVTVTV, KVVVVVVVVM, IKTKTKVRVV, VDVDVDVTVT, TVKVDVDVDV, ILKLELTLTL, DTDTVTVTVT, KSVDVDVDTV, VTVTVTHTHT, RVTVTVTVTH, STQITITITI, TLQATATATA, KTTHTHTHTH, ETETETETEK, DSEFTETETE, TITIDIDIDI, VIVIVIVIVI, TALVIVIVIV, LILILILSLT, TFTLLLILIL, LILILILIVK, TVTLILILIL, TLKATATATA, TVTVVIVIVI, VKVKVEVEVE, VIKVKVKVKV, TKKKTNTNTN, TVKIKITITI, SN TTTTTTTT, ATATATATAK, TVTVTAVATA, SKAQTHTHTH, ITITFTFTVT, TITITITFTF, TKNNVNVNTN, NTNTHTHTHT, TKTNTNTHTH, TTTKTQTQTQ, IITVTATATA, DIDIDIDIDI, TTIDIDIDID, SKTNTNTNTN, NISVSVTVTV, KFKLTLTLTL, TVEVSVSVSV, ATATATAVAV,

[0517]

[0518] VVTYTATATA, NININININN, TKENINININ, VTATATATVK, RAEFTFTFTF, QLSVTVIVTV, VKEKTHTHTH, IIEITITITI, ILKITITITI, KEEKTKTKTK, TLTQTQTQTQ, LTFTFTFTFT, NTRSTSTSTS, STTETKTKTK, IEEETETETE, VVVTVTVTVT, KNKNKVKNVN, EAERTKNKNK, DVSV IIIIII, TFEVTVTVTV, FEFEFEFEFK, KVEFEFEFEF, ITITITIIIK, TATITITITI, KITITITATA, TVKVDADATA, TVKVVVTVTV, TVNVVVVNVV, NTITITITVT, TVNVTVTITV, KTEITITITI, IEIEIKIKII, TIEIEIEIKI, YTYTYVYTIT, TVEYVYVYTY, IQILILISIS, QIKILILILI, TLTLVLVLVL, VTATFTATAT, VAKATATATA, EVELTVTVTV, TVTITTTLTT,

[0519] TTTTTTTT DT, TTTL IIIIII, VQVQIQVQVQ, TITITIQVQV, KVEVKATATA, TLTLTLSLTL, TTLTLTLTLT, IIANININLN, TASINININI, VKEDIKIKIK, KITVTVTVTV, VTLTLTVTVT, TVQVTVIVTV, AVAVAVAVVT, TVELVAVAVA, TITLTLTVTV, TVTLTLTITI, STNTNTNTNT, NTSNTNTNTN, RVTVTFTFTF, TKEETETETE, ITITVTVTVT, TITGTVTVTV, VVVVVVVVVV, DVKVVVVVVV, DVKITITITI, EVEVVVVVVV, KTKTITKTKK, ITITITATAT, TAKVTVTVTA, TETETNTNTN, KVKVTITITI, ITITITITIN, NISITITITL, IKEVVIVIVI, KVTLTLTVTV, VIEITVTVTV, IARVTITITI, QVQVQVQTQT, TTKTVTVQVQ, TVEITITITV, KDKTKTKTKT, TTTDTDTDTK, EVKVTVTLTV, TDRSVSLNTN, SRSANVQSVT, VTTIVLVVNL, SNSTVTLQQT, STTIVLTVNL, ITLVLVQSTT, VVKVTTTTKV, MSVTTTVVVS, SVTVVATTTV, VTVTTVVQST, TEKIKKEYAD,SNSEKEVVKT, TEKTEKEYED, NGIEKVVNVT, QIRRSYKYKN, RSSESERQTT, TSTEEYEYNN, RRIKYTRTTT, QLRVEVTVSV, STSVVRVERR, SRRVVVRVNV, VTVTVERRET, ILRVEINETD, SQSRIEIEYV, KVRVEIEEND, VRINIEYELT, VLTNTNTNKE, SSNTNTNTIV, KRTNTNTNNE, NTNTNTNSTT, ALKSVKVDTN, STSKDEKEVK, TVTQVKEDDN, SKKVDEKTIT, VVRVAVAVTV, SSITVEVAVA, TVTIHVEVNV, VDVAVAAQVT, KNTSTNTYSL, SKSENKNTTT, SQTSENKYSL, STSTNTNQTT, VDKVEIEVKV, SEITVKNEHN, SNKKKIKVTV, VTIENENKVT, VDTNNNNYTI, SNSNYRNTNN, TRTDENRYNI, YDNNYTNTRT, VK I I I I I VSV, STVTVTTTQQ, SSIIIIIVNV, V I I I I I QKQT, VDKIEVDHSL, SNSSHETTKT, KTTKKEEHDL, IKIDVETTVT, VNEVTITVNI, SNSEVKIRKV, TETVVIKVNI, VEITITTTVT, VLSVSVSARV, VDVQVEVQQT, TTRTEVEANV, VEVSVSQQVT, VARTSVSLTA, STLEVEEERA, QRRTTIELNA, TTVSVEESVT, VVTVTVTYTD, SSVTVTVTTT, STTVSVTYND, VTVTVTVSTT, VDTVSISYNV, LNVSVSQQQK, SQTVSISYNV, VTVSIQQTVT, TTRVQVQVTN, STTTVQDEDQ, SKRVQVQVSN, VTVQNEDLTT, VLEVEVEVKV, VEIVVVVKNT, KSEVKVVVNV, VSIEVNKQVT, VHSINVNVTV, STINVNVTNT, RSTININVNV, ISVNVTVSVT, KNTVTKTVDL, AREVVVKTTT, TKTETKVVDL, and VTKTVTTTQT. The one or more beta strands and / or one or more alpha helices may comprise one or more of the listed insertions. Different combination of these ten amino acid elongations / insertions may be used in the same beta strand or alpha helix or in different combinations of two or more beta strands and / or two or more alpha helices. The ten amino acid elongations / insertions may be used in the one or more beta strands and / or one or more alpha helices consecutively or separated by existing sequences.

[0520] The one or more beta strands and / or one or more alpha helices may be elongated by one or more of TNVNTSTSTYNI, TTSTSTTTFEVT, SQRNSITSTYNI, STNTSSSTFMQR, TSQTRTN ITYSV, GSTDVRIEQKQQ, SNTTETRIRYTV, VTTEVNIKQTQK, TTTQTQTQSTN L, SNAENKQKQTQN, TQTQEQKQKTNL, ATQTSTQTQRQQ, TQTNVNVTGNTN, SSSTNMNTNQLT, TQTNTNTNNNGN, NEIVNVNQNQIK, VQTQTNTNSVSV, SNSTTTNVNTTS, TNSQVNVNSVNV, STNTNTNTNITR, VQKVSNSNTNTN, SNSKNETEHTMI, TTTVTNENENTN, NSNSNSIMHTVK, KTVIKQEQDQLV, SNQVQRVEQENV, SNKVKQRQVQSV, QIQKQEQEILVK, QNTQTNTYTYTG, STNTSTYKQEIK, SQKQENKYTYSG, NTTTYTYEQSVR, VVTLTVTTTVTL, STVSVTTTVNTT, SVTLTVTTTVTL, VTL I I I I I VTTR, VNTITVTVTLTV, STSSTSVKVNQQ, SNNITVKVSLTV, VNITVTVNNQQR, VQTHAYQLTHTV, SNSRVELIQQTV, SQTTTYKLEHTV, YKHTYQLQQTVK, KVEIKINIDTNV, SDIDIIYQTSTT, SVKTEIQIDTNV, INIDINVKTSTK, KVSNKVKVKITA, SNIGVTHEVTTT, TNRNTVEVDINA, VNIKVKHTVETR, TNTITVTYTYTN, SSVTVTNQVTVV, STTIQVTYTYNN, VTVTYTQTVSTK, KTTIIVVVEVEN, SKVEVEVETTTI, SNTINVEVEVEN, VKIIVVVTTQVK, TQKVAIAVNVHV, SNVEQVLENTNT, TNTVEIELVVNV, VTVAVAITNTTK, VSSITVTSTVEL, SKVTVTVTNVTT, SQSNTVTVTVEL, VTVTVTSVNTVR, TNTTNVTINVTL, ATTIVIYNYTTT, TVTTNVNIIVNL, VITNVNYTNTTK, IVTVTVTVNNTV, SSSTVTVTAAVS, TTTVTTTVTN DV, and VTTTVTVTATVK. The one or more beta strands and / or one or more alpha helices may comprise one or more of the listed insertions. Different combination of these twelve aminoacid elongations / insertions may be used in the same beta strand or alpha helix or different combinations of two or more beta strands and / or two or more alpha helices. The twelve amino acid elongations / insertions may be used in the one or more beta strands and / or one or more alpha helices consecutively or separated by existing sequences.

[0521] The one or more beta strands and / or one or more alpha helices may be elongated by one or more of INRNVNVNVNVYET, SKSENKNEIENVTR, TQKNKNEIENKYET, SRNVNVNVNTNEVK, VVINENQNVNNYNV, SNNLNQNQNRVETT, TTKNENQNQNEYNV, NNNKNVNRNIVTVK, VNTNTNTNSNDYSV, SNSNNTNTNTLTTS, SQTNINTATNDYDV, INNTNTNTNTHTVK, QNTIVTVTVTVTSV, SKSVTVVTTQNTNP, RNSTQTTTVTVTSV, SVTVTVTVTQNSTR, KNSNTNTNTVVYTA, STLTVTTTNANNNS, SNKTASANTVTYTA, VTITNTNTNNNETK, VITNTNTNENTYKN, SESNNTNTNTVTVK, SVENTNTNTNNYTN, SKNTNTNENTVIVR, IVQTETETEVTVLV, STTSTSNETSVTIR, STRTSTETEVSVSV, TNTETSVETSVGVK, VVTTKVIVLTTYAN, NRNEVKAKVEVTQR, TTKTTVSVKTKYSN, NTVITLVIVEVEQR, ILKTQQKQKVNINI, SNYTVEVEQEQQVT, KVSTSQEQEVRINI, IEVQVEQKQQQEVR, VLSVEVSTSNDYNI, SNSKVDNDVRVKTT, TNEVIVDVDNKYNI, SSNSVSTSVRVKTK, LVLIQNTQTHTVSV, SSYTYTTTQTVLLQ, TTAITNTQTHTVSV, YTYTQTTTQLVVQR, VAKVTNTVTNTYSI, SKISITNTVTSTLN, RQTVTHTVTNTYSI, ISNTVTNTVTVSVK, VVEISVNNNVSYRN, SNSTNSVSNTVKVT, SVSIKVTNSVSYSN, and STVSNNVNVSVSVK. The one or more beta strands and / or one or more alpha helices may comprise one or more of the listed insertions. Different combination of these fourteen amino acid elongations / insertions may be used in the same beta strand or alpha helix or different combinations of two or more beta strands and / or two or more alpha helices. The fourteen amino acid elongations / insertions may be used in the one or more beta strands and / or one or more alpha helices consecutively or separated by existing sequences.

[0522] All of the lists of preferred amino acids above are particularly preferred for modifications, elongations or insertions in one or more beta-strands.

[0523] The pore monomer preferably comprises four beta-strands. At least two of the four betstrands may be modified or elongated. At least three of the four bet-strands may be modified or elongated. Preferably, all of the four beta-strands are modified or elongated. The four beta-strands may be modified or elongated to different degrees. The four betastrands are preferably modified or elongated to the same degree. In this context, the term "degree" typically relates to the number of amino acids added to the two or more betastrands. For instance, two amino acids may be added to each of the four beta-strands.

[0524] Even if the four beta-strands are modified or elongated to the same degree, they may be modified in different ways. For instance, the two amino acids added to each of the four beta-strands may be different. The two amino acids may be selected from the list above.Alternatively, four consecutive amino acids may be added to two of the beta-strands and two separate stretches of two amino acids may be added to the other two beta-strands. The identities of the four consecutive amino acids and the two separate stretches of two amino acids may be the same or different. The skilled person is capable of designing suitable modifications and elongations / insertions.

[0525] Each beta-strand may comprise one or more preferred positions for modifications, elongations or insertions. Each beta-strand may comprise at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9 or at least about 10 preferred positions for modifications, elongations or insertions. The preferred positions in each of the four beta-strands typically line up approximately horizontally in the beta-sheet formed from the four beta-strands.

[0526] Table 3 below shows a pore monomer comprising four beta-strands capable of forming a pore comprising a beta-barrel channel. Each strand comprises four preferred positions for modifications, elongations or insertions (labelled A-D, E-H, I-L and M-P). The preferred positions in each row, e.g., A, E, I and M, typically line up approximately horizontally in the beta-sheet formed by the four beta-strands.

[0527] Beta-strand 1 Beta-strand 2 Beta-strand 3 Beta-strand 4

[0528] A E I M

[0529] B F J N

[0530] C G K 0

[0531] D H L P

[0532]

[0533] The modified pore monomer may comprise any of the following modifications, elongations or insertions based on Table 3:

[0534] • The insertion of the same number of consecutive amino acids, such as 2, 4, 6, 8, or 10 consecutive amino acids, in beta-strands 1-4 at the positions in the same row, such as (i) A, E, I and M, (ii) B, F, J and N, (iii) C, G, K and O or (iv) D, H, L and P.

[0535] • The insertion of the same number of consecutive amino acids, such as 2, 4, 6, 8, or 10 consecutive amino acids, at any one of positions A-D in beta-strand 1, at any one of positions E-H in beta-strand 2, at any one of positions I-L in beta-strand 3, and at any one of positions M-P in beta-strand 4.• The insertion of the same number of amino acids, such as 2, 4, 6, 8, or 10 amino acids, in each of beta-strands 1-4, wherein the same number of amino acids are consecutive amino acids inserted at any one position in a strand and / or the same number of amino acids are two or more stretches of one or more amino acids inserted at two or more positions in a strand. The two or more stretches may be 2, 3, 4, 5 or more stretches. The two or more stretches may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids.

[0536] In any of the embodiments, the inserted amino acids, consecutive amino acids and / or two or more stretches of one or more amino acids may be selected any of the individual amino acids or stretches of 2, 4, 6, 8 or 10 amino acids listed above.

[0537] The pore monomer preferably comprises two beta-strands. At least one of the two betstrands may be modified or elongated. Both of the two beta-strands may be modified or elongated. The two beta-strands may be modified or elongated to different degrees. The two beta-strands are preferably modified or elongated to the same degree. In this context, the term "degree" typically relates to the number of amino acids added to the two betastrands. For instance, two amino acids may be added to each of the two beta-strands. Even if the two beta-strands are modified or elongated to the same degree, they may be modified in different ways. For instance, two amino acids added to each of the two betastrands may be different. The two amino acids may be selected from the list above.

[0538] Alternatively, four consecutive amino acids may be added to one beta-strand and two separate stretches of two amino acids may be added to the other beta-strand. The identities of the four consecutive amino acids and the two separate stretches of two amino acids may be the same or different. The skilled person is capable of designing suitable modifications and elongations / insertions.

[0539] Each beta-strand may comprise one or more preferred positions for modifications, elongations or insertions. Each beta-strand may comprise at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9 or at least about 10 preferred positions for modifications, elongations or insertions. The preferred positions in each of the two beta-strands typically line up approximately horizontally in the beta-sheet formed from the two beta-strands.

[0540] Table 4 below shows a pore monomer comprising two beta-strands capable of forming a pore comprising a beta-barrel channel. Each strand comprises four preferred positions for modifications, elongations or insertions (labelled A-D and E-H). The preferred positions in each row, e.g., A and E, typically line up approximately horizontally in the beta-sheet formed by the two beta-strands.Beta-strand 1 Beta-strand 2

[0541] A E

[0542] B F

[0543] C G

[0544] D H

[0545]

[0546] The modified pore monomer may comprise any of the following modifications, elongations or insertions based on Table 4:

[0547] • The insertion of the same number of consecutive amino acids, such as 2, 4, 6, 8, or 10 consecutive amino acids, in both beta-strands 1-4 at the positions in the same row, such as (i) A and E, (ii) B and F, (iii) C and G or (iv) D and H.

[0548] • The insertion of the same number of consecutive amino acids, such as 2, 4, 6, 8, or 10 consecutive amino acids, at any one of positions A-D in beta-strand 1, and at any one of positions E-H in beta-strand 2.

[0549] • The insertion of the same number of amino acids, such as 2, 4, 6, 8, or 10 amino acids, in each beta-strand, wherein the same number of amino acids are consecutive amino acids inserted at any one position in a strand and / or the same number of amino acids are two or more stretches of one or more amino acids inserted at two or more positions in a strand. The two or more stretches may be 2, 3, 4, 5 or more stretches. The two or more stretches may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids.

[0550] In any of the embodiments, the inserted amino acids, consecutive amino acids and / or two or more stretches of one or more amino acids may be selected any of the individual amino acids or stretches of 2, 4, 6, 8 or 10 amino acids listed above.

[0551] The modified pore monomer is preferably not modified to alter the length of the constriction region. The modified pore monomer is preferably not modified using a spacer above the surface region to increase the distance between an enzyme docking site and the constriction region.

[0552] Preferred modified pore monomersThe modified pore monomer is preferably a structural variant having a root mean square deviation (RMSD) of less than about 4.0 Angstroms (A), such as less than about 3.5 A, less than about 3.0 A, less than about 2.5 A, less than about 2.0 A, less than about 1.5 A, less than about 1.0 A or less than about 0.5 A, when compared with a protein having the sequence shown any one of SEQ ID NOs: 13-21.

[0553] The invention also provides a modified actinobacterial pore monomer having a root mean square deviation (RMSD) of less than about 4.0 A such as less than about 3.5 A, less than about 3.0 A, less than about 2.5 A, less than about 2.0 A, less than about 1.5 A, less than about 1.0 A or less than about 0.5 A, when compared with a protein having the sequence shown in any one of SEQ ID NOs: 13-21.

[0554] The modified pore monomer preferably comprises a variant sequence having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity to the sequence of any one of SEQ ID NOs: 13-21. The pore monomer or modified pore monomer preferably comprises a variant sequence having at least about 80% homology and / or identity to the sequence of any one of SEQ ID NOs: 13-21. The pore monomer or modified pore monomer preferably comprises a sequence having at least about 90% homology and / or identity to the sequence of any one of SEQ ID NOs: 13-21. The pore monomer preferably comprises a sequence having 100% homology and / or identity to the sequence of any one of SEQ ID NOs: 13-21. Homology and / or identity is typically measured over the entire length of the monomer.

[0555] Sequence homology and / or identity can also relate to a fragment or portion of the reference sequence. Hence, a sequence may have less than 40% overall sequence homology and / or identity with any one of SEQ ID NOs: 13-21, but the sequence of a particular region, domain or subunit could share at least about 80%, at least about 90%, or as much as about 99% sequence homology and / or identity with the corresponding region of the wild-type or native monomer. There may be at least about 80%, at least 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98% or at least about 99% homology and / or identity over a stretch of 100 or more, for example 125, 150, 175 or 200 or more, contiguous amino acids ("hard homology").

[0556] The invention also provides a modified actinobacterial pore monomer comprising a sequence having at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about97%, at least about 98% or at least about 99% homology and / or identity to the sequence of any one of SEQ ID NOs: 13-21.

[0557] Modified actinobacterial pore monomers with specific lengths

[0558] The invention also provides modified actinobacterial pore monomer comprising (a) a cap region which comprises a surface region that forms part of the upper surface of a pore formed from the modified pore monomer and (b) a constriction region that forms part of a constriction in a channel of a pore formed from the modified pore monomer, wherein the distance between the surface region and the constriction region is at least 79 angstroms (Å). All of the discussion above concerning cap regions, surface regions, upper surfaces, constriction regions, constrictions and channels equally apply to these modified pore monomers. The actinobacterial pore monomer may be any of those discussed above, including a modified PorARc monomer or a modified MspA pore monomer.

[0559] The distance is preferably increased to from about 79 A to about 200 A, such as from about 80 A to about 180 A, from about 90 A to about 160 A, or from about 100 A to about 200 A. The distance may be increased to about or at least about 79 A, about or at least about 80 A, about or at least about 81 A, about or at least about 82 A, about or at least about 83 A, about or at least about 84 A, about or at least about 85 A, about or at least about 86 A, about or at least about 87 A, about or at least about 88 A, about or at least about 89 A, about or at least about 90 A, about or at least about 91 A, about or at least about 92 A, about or at least about 93 A, about or at least about 94 A, about or at least about 95 A, about or at least about 96 A, about or at least about 97 A, about or at least about 98 A, about or at least about 99 A, about or at least about 100 A, about or at least about 101 A, about or at least about 102 A, about or at least about 103 A, about or at least about 104 A, about or at least about 105 A, about or at least about 106 A, about or at least about 107 A, about or at least about 108 A, about or at least about 109 A, about or at least about 110 A, about or at least about 111 A, about or at least about 112 A, about or at least about 113 A, about or at least about 114 A, about or at least about 115 A, about or at least about 116 A, about or at least about 117 A, about or at least about 118 A, about or at least about 119 A, about or at least about 120 A, about or at least about 121 A, about or at least about 122 A, about or at least about 123 A, about or at least about 124 A, about or at least about 125 A, about or at least about 126 A, about or at least about 127 A, about or at least about 128 A, about or at least about 129 A, about or at least about 130 A, about or at least about 131 A, about or at least about 132 A, about or at least about 133 A, about or at least about 134 A, about or at least about 135 A, about or at least about 136 A, about or at least about 137 A, about or at least about 138 A, about or at least about 139 A, about or at least about 140 A, about or at least about 141 A, about or at least about 142 A, about or at least about 143 A, about or at least about 144 A, about or at least about 145 A, about or at least about 146 A,about or at least about 147 A, about or at least about 148 A, about or at least about 149 A, about or at least about 150 A, about or at least about 151 A, about or at least about 152 A, about or at least about 153 A, about or at least about 154 A, about or at least about 155 A, about or at least about 156 A, about or at least about 157 A, about or at least about 158 A, about or at least about 159 A, about or at least about 160 A, about or at least about 161 A, about or at least about 162 A, about or at least about 163 A, about or at least about 164 A, about or at least about 165 A, about or at least about 166 A, about or at least about 167 A, about or at least about 168 A, about or at least about 169 A, about or at least about 170 A, about or at least about 171 A, about or at least about 172 A, about or at least about 173 Å, about or at least about 174 Å, about or at least about 175 Å, about or at least about 176 Å, about or at least about 177 Å, about or at least about 178 Å, about or at least about 179 Å, about or at least about 180 Å, about or at least about 181 Å, about or at least about 182 Å, about or at least about 183 Å, about or at least about 184 Å, about or at least about 185 Å, about or at least about 186 Å, about or at least about 187 Å, about or at least about 188 Å, about or at least about 189 Å, about or at least about 190 Å, about or at least about 191 Å, about or at least about 192 Å, about or at least about 193 Å, about or at least about 194 Å, about or at least about 195 Å, about or at least about 196 Å, about or at least about 197 Å, about or at least about 198 Å, about or at least about 199 Å or about or at least about 200 Å.

[0560] Constructs

[0561] The invention also provides a construct comprising two or more covalently attached modified pore monomers of the invention.

[0562] The construct may comprise 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more or 10 or more modified pore monomers of the invention. The construct may comprise at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 modified pore monomers of the invention.

[0563] The two or more modified pore monomers of the invention may be the same or different. The two or more modified pore monomers may differ based on one or more of (a) the sequence of the pore monomer, (b) the length of the elongation / insertion, (c) the amino acid(s) elongation / insertion, (d) the format of the elongation / insertion (e.g., consecutive versus separated), and (e) other modifications of the pore monomer. The modified pore monomers may differ based on (a); (b); (c); (d); (e); (a) and (b); (a) and (c); (a) and (d); (a) and (e); (b) and (c); (b) and (d); (b) and (e); (c) and (d); (c) and (e); (d) and (e); (a), (b) and (c); (a), (b) and (d); (a), (b) and (e); (a), (c) and (d); (a), (c) and (e); (a), (d) and (e); (b), (c) and (d); (b), (c) and (e); (b), (d) and (e); (c), (d) and (e); (a), (b), (c) and (d); (a), (b), (c) and (e); (a), (b), (d) and (e); (a), (c), (d) and (e); (b), (c), (d) and (e);and (a), (b), (c), (d) and (e). The two or more pore monomers are preferably the same (*i.e.*, identical).

[0564] The construct preferably comprises two modified pore monomers. The two modified pore monomers may be the same or different. The two modified pore monomers are preferably the same (*i.e.*, identical).

[0565] The modified pore monomers may be genetically fused, optionally via a linker, or chemically fused, for instance via a chemical crosslinker. Methods for covalently attaching monomers are disclosed in WO 2017 / 149316, WO 2017 / 149317, and WO 2017 / 149318 (incorporated herein by reference in their entirety). The modified pore monomers may be genetically fused.

[0566] The linker is preferably an amino acid sequence and / or a chemical crosslinker. Suitable amino acid linkers, such as peptide linkers, are known in the art. The length, flexibility and hydrophilicity of the amino acid or peptide linker are typically designed such that the modified pore monomers are capable of forming a pore. Preferred flexible peptide linkers are stretches of 2 to 20, such as 4, 6, 8, 10 or 16, serine and / or glycine amino acids. More preferred flexible linkers include (SG)1, (SG)2, (SG)3, (SG)4, (SG)5, (SG)8, (SG)10, (SG)15or (SG)20wherein S is serine and G is glycine. Preferred rigid linkers are stretches of 2 to 30, such as 4, 6, 8, 16 or 24, proline amino acids. More preferred rigid linkers include (P)12wherein P is proline.

[0567] Suitable chemical crosslinkers are well-known in the art. Suitable chemical crosslinkers include, but are not limited to, those including the following functional groups: maleimide, active esters, succinimide, azide, alkyne (such as dibenzocyclooctynol (DIBO or DBCO), difluoro cycloalkynes and linear alkynes), phosphine (such as those used in traceless and non-traceless...

Claims

CLAIMS1. A modified actinobacterial pore monomer comprising (a) a cap region which comprises a surface region that forms part of the upper surface of a pore formed from the modified actinobacterial pore monomer and (b) a constriction region that forms part of a constriction in a channel of a pore formed from the modified actinobacterial pore monomer, wherein the actinobacterial pore monomer is modified to increase the distance between the surface region and the constriction region.

2. A modified actinobacterial monomer according to claim 1, wherein the distance between the surface region and the constriction region is increased by about 4 or more angstroms (A) and / or by the length of about 1 or more nucleotides or amino acids.

3. A modified actinobacterial pore monomer comprising (a) a cap region which comprises a surface region that forms part of the upper surface of a pore formed from the modified actinobacterial pore monomer and (b) a constriction region that forms part of a constriction in a channel of a pore formed from the modified actinobacterial pore monomer, wherein the distance between the surface region and the constriction region is at least 79 angstroms (Å).

4. A modified actinobacterial pore monomer according to any one of the preceding claims, wherein the modified actinobacterial pore monomer further comprises a transmembrane region.

5. A modified actinobacterial pore monomer according to claim 4, wherein the transmembrane region comprises one or more beta-strands.

6. A modified actinobacterial pore monomer according to claim 5, wherein the one or more beta-strands are modified.

7. A modified actinobacterial pore monomer according to claim 6, wherein the one or more beta-strands are elongated.

8. A modified actinobacterial pore monomer according to claim 7, wherein the one or more beta-strands are elongated by the addition of from about 1 to about 24 amino acids.

9. A modified actinobacterial pore monomer according to claim 8, wherein the amino acid(s) is / are selected from tyrosine (Y), arginine (R), lysine (K), phenylalanine (F), tryptophan (W), threonine (T), valine (V), alanine (A), isoleucine (I), leucine (L), glycine (G), glutamic acid (E), aspartic acid (D), histidine (H), methionine (M), cysteine (C), proline (P), serine (S), asparagine (N), glutamine (Q) and unnatural amino acids.

10. A modified actinobacterial pore monomer according to claim 8 or 9, wherein the outward facing amino acid(s) are hydrophobic and are located within the membranespanning region of the transmembrane region.

11. A modified actinobacterial pore monomer according to claim 8 or 9, wherein the amino acid(s) are located outside of the membrane-spanning region of the transmembrane region.

12. A modified actinobacterial pore monomer according to any one of the preceding claims, wherein the modified actinobacterial pore monomer is not modified to alter the length of the constriction region.

13. A modified actinobacterial pore monomer according to any one of the preceding claims, wherein the modified actinobacterial pore monomer is from PorARc, PorARr or PorBRr.

14. A modified actinobacterial pore monomer according to any one of claims 1-12, wherein the modified actinobacterial pore monomer is from MspA, MspB, MspC or MspD.

15. A modified actinobacterial pore monomer according to any one of the preceding claims, wherein the modified actinobacterial pore monomer comprises a chimeric pore monomer comprising two or more regions, wherein at least two of the two or more regions are from different actinobacterial pores.

16. A construct comprising two or more covalently attached modified actinobacterial pore monomers according to any one of the preceding claims.

17. A pore comprising at least one modified actinobacterial pore monomer according to any one of claims 1-15.

18. A pore comprising at least one construct according to claim 16.

19. An oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, wherein each modified actinobacterial pore monomer comprises (a) a cap region which comprises a surface region forming part of the upper surface of the oligomeric pore and (b) a constriction region forming part of a constriction in the channel, and wherein each actinobacterial pore monomer is modified to increase the distance between the upper surface of the oligomeric pore and the constriction.

20. An oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, wherein each modified actinobacterial pore monomer comprises (a) a cap region which comprises a surface region forming part of the upper surface of theoligomeric pore, (b) a transmembrane region forming the channel and (c) a constriction region forming part of a constriction in the channel, and wherein the transmembrane region in each actinobacterial pore monomer is modified to increase the distance between the upper surface of the oligomeric pore and the constriction.

21. An oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, wherein each modified actinobacterial pore monomer comprises (a) a cap region which comprises a surface region forming part of the upper surface of the oligomeric pore and (b) a constriction region forming part of a constriction in the channel, and wherein the distance between the upper surface of the oligomeric pore and the constriction is at least 79 angstroms (Å).

22. A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel and (2) an analyte binding protein, wherein the oligomeric pore comprises (a) a cap region which comprises an upper surface which interacts with the analyte binding protein and (b) a constriction in the channel, and wherein each actinobacterial pore monomer is modified to increase the distance between the upper surface and the constriction.

23. A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel and (2) an analyte binding protein, wherein the oligomeric pore comprises (a) a cap region which comprises an upper surface which interacts with the analyte binding protein, (b) a transmembrane region forming the channel and (c) a constriction in the channel, and wherein the transmembrane region in each actinobacterial pore monomer is modified to increase the distance between the upper surface and the constriction.

24. A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel and (2) an analyte binding protein, wherein the oligomeric pore comprises (a) a cap region which comprises an upper surface which interacts with the analyte binding protein and (b) a constriction in the channel, and wherein the distance between the upper surface and the constriction is at least 79 angstroms (Å).

25. A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, (2) an analyte present in the channel of the pore, and (3) an analyte binding protein bound to the analyte, wherein the oligomeric pore comprises (a) a cap region which prevents the analyte binding protein from entering the channel and (b) a constriction in the channel, and wherein each actinobacterial pore monomer is modified to increase the minimum distance between the analyte binding protein and the constriction.

26. A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, (2) an analyte present in the channel of the pore, and (3) an analyte binding protein bound to the analyte, wherein the oligomeric pore comprises (a) a cap region which prevents the analyte binding protein from entering the channel, (b) a transmembrane region forming the channel and (c) a constriction in the channel, and wherein the transmembrane region in each actinobacterial pore monomer is modified to increase the minimum distance between the analyte binding protein and the constriction.

27. A pore complex comprising (1) an oligomeric pore formed from modified actinobacterial pore monomers surrounding a channel, (2) an analyte present in the channel of the pore, and (3) an analyte binding protein bound to the analyte, wherein the oligomeric pore comprises (a) a cap region which prevents the analyte binding protein from entering the channel and (b) a constriction in the channel, and wherein the minimum distance between the analyte binding protein and the constriction is at least 79 angstroms (Å).

28. A pore according to claim 17 or 18, an oligomeric pore according to any one of claims 19-21 or a pore complex according to any one of claims 22-27, wherein the pore, oligomeric pore or pore complex is / comprises a homooligomer comprising 3 to 24 modified actinobacterial pore monomers.

29. A membrane comprising a pore according to claim 17 or 18, an oligomeric pore according to any one of claims 19-21 or a pore complex according to any one of claims 22-27.

30. A method for determining the presence, absence or one or more characteristics of a target analyte, comprising the steps of: (i) contacting the target analyte with a pore according to claim 17 or 18, an oligomeric pore according to any one of claims 19-21 or a pore complex according to any one of claims 22-27, such that the target analyte moves with respect to the pore, oligomeric pore or pore complex; and (ii) taking one or more measurements as the analyte moves with respect to the pore, oligomeric pore or pore complex and thereby determining the presence, absence or one or more characteristics of the analyte.

31. A method according to claim 30, wherein the target analyte is a metal ion, a volatile or non-volatile organic compound an inorganic salt, a polymer, an amino acid, a peptide, a polypeptide, a protein, a nucleotide, an oligonucleotide, a polynucleotide, a polynucleotide-polypeptide conjugate, a monosaccharide, an oligosaccharide, or a polysaccharide.

32. A method according to claim 30 or 31, wherein the method comprises determining one or more characteristics selected from (i) the length of the target analyte, (ii) the identity of the target analyte, (iii) the sequence of the target analyte, (iv) the secondary structure of the target analyte and (v) whether or not the target analyte is modified.

33. Use of a pore according to claim 17 or 18, an oligomeric pore according to any one of claims 19-21 or a pore complex according to any one of claims 22-27 to determine the presence, absence or one or more characteristics of a target analyte.

34. A polynucleotide which encodes a modified actinobacterial pore monomer according to any one of claims 1-15 or a construct according to claim 16.

35. A kit for characterising a target analyte comprising (a) a pore according to claim 17 or 18, an oligomeric pore according to any one of claims 19-21 or a pore complex according to any one of claims 22-27 and (b) the components of a membrane.

36. A kit for characterising a target polynucleotide or a target polypeptide comprising (a) a pore according to claim 17 or 18 or an oligomeric pore according to any one of claims 19-21 and (b) an analyte binding protein.

37. An apparatus for characterising a target analyte in a sample, comprising (a) a plurality of pores according to claim 17 or 18 or a plurality of oligomeric pores according to any one of claims 19-21 and (b) a plurality of analyte binding proteins.

38. An array comprising a plurality of membranes according to claim 29.

39. A system comprising (a) a membrane according to claim 29 or an array according to claim 38, (b) means for applying a potential across the membrane(s) and (c) means for detecting electrical or optical signals across the membrane(s).

40. An apparatus comprising a pore according to claim 17 or 18, an oligomeric pore according to any one of claims 19-21 or a pore complex according to any one of claims 22-27 inserted into an in vitro membrane.

41. An apparatus produced by a method comprising (i) obtaining a pore according to claim 17 or 18, an oligomeric pore according to any one of claims 19-21 or a pore complex according to any one of claims 22-27 and (ii) contacting the pore, pore complex, oligomeric pore or oligomeric pore complex with an in vitro membrane such that the pore, pore complex, oligomeric pore or oligomeric pore complex is inserted in the in vitro membrane.

42. A method of improving the ability of an oligomeric pore formed from actinobacterial pore monomers surrounding a channel to characterise an analyte, wherein each actinobacterial pore monomer comprises (a) a cap region which comprises a surface region forming part of the upper surface of the oligomeric pore and (b) a constriction region forming a constriction in the channel, the method comprising modifying at least one of the actinobacterial pore monomers to increase the distance between the upper surface and the constriction.

43. A method of improving the ability of a pore complex comprising (1) an oligomeric pore formed from actinobacterial pore monomers surrounding a channel and (2) an analyte binding protein to characterise an analyte, wherein each actinobacterial pore monomer comprises (a) a cap region which comprises an upper surface which interacts with the analyte binding protein and (b) a constriction in the channel, the method comprising modifying at least one of the actinobacterial pore monomers to increase the distance between the upper surface and the constriction.

44. A method of improving the ability of a pore complex comprising (1) an oligomeric pore formed from actinobacterial pore monomers surrounding a channel, (2) an analyte present in the channel of the pore, and (3) an analyte binding protein bound to the analyte, wherein the oligomeric pore comprises (a) a cap region which prevents the analyte binding protein from entering the channel and (b) a constriction in the channel, the method comprising modifying at least one of the actinobacterial pore monomers to increase the distance between the minimum analyte binding protein and the constriction.

45. A modified actinobacterial pore monomer having a root mean square deviation (RMSD) of less than about 4.0 Å when compared with a protein having the sequence shown in any one of SEQ ID NOs: 13-21.

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