LEADER SEQUENCE
20230076614 · 2023-03-09
Assignee
Inventors
Cpc classification
B82Y5/00
PERFORMING OPERATIONS; TRANSPORTING
C07K2319/035
CHEMISTRY; METALLURGY
International classification
Abstract
The present invention provides use of a Photorhabdus Virulence Cassettes (PVC) effector leader sequence, for packaging a payload into a PVC Needle Complex, and related methods for manufacturing a packaged PVC Needle Complex. The payload is one or more selected from a polypeptide, a nucleic acid, or a combination thereof, and the leader sequence and the payload form an effector fusion that is distinct from a wild-type PVC effector protein.
Claims
1. A method for packaging a payload into a Photorhabdus Virulence Cassettes (PVC) Needle Complex, the method comprising: a. providing an effector fusion comprising a PVC effector leader sequence fused to a payload, and b. contacting a PVC Needle Complex with said fusion thereby allowing the leader sequence to package the payload into the Needle Complex; wherein the payload is one or more selected from a polypeptide, a nucleic acid, or a combination thereof; and wherein the leader sequence and the payload form an effector fusion that is distinct from a wild-type PVC effector protein.
2. The method according to claim 1, wherein the leader sequence comprises amino acid residues 1-50 of a PVC effector.
3. The method use according to claim 1, wherein the leader sequence comprises an amino acid sequence having at least 60% sequence identity to one or more sequence selected sequence from SEQ ID NO.: 47-SEQ ID NO.: 92.
4. The method according to claim 1, wherein the PVC effector comprises an amino acid sequence of one or more sequence selected from SEQ ID NO.: 1-SEQ ID NO.: 46.
5. The method use according to claim 1, wherein the PVC effector comprises a sequence selected from SEQ ID NO: 4, SEQ ID NO: 22, SEQ ID NO: 25, SEQ ID NO: 30, SEQ ID NO: 32 and SEQ ID NO: 46.
6. The method use according to claim 1, wherein the leader sequence is covalently fused to the payload, preferably at an N-terminus of the payload.
7. (canceled)
8. The method according to claim 1, wherein said contacting occurs within a cell, in a cell lysate, or in a purified cell lysate.
9. The method of claim 1, wherein said method is performed in vitro and/or ex vivo.
10. (canceled)
11. (canceled)
12. A PVC Needle Complex comprising an effector fusion; a. wherein said effector fusion comprises a PVC effector leader sequence fused to a payload; b. wherein said payload is one or more selected from a polypeptide, a nucleic acid or a combination thereof; and c. wherein the effector fusion is distinct from a wild-type PVC effector protein.
13. An effector fusion, comprising a PVC effector leader sequence fused to a payload; a. wherein said payload is one or more selected from a polypeptide, a nucleic acid or a combination thereof; and b. wherein the effector fusion is distinct from a wild-type PVC effector protein.
14. (canceled)
15. The PVC Needle Complex according to claim 12, wherein the leader sequence comprises amino acid residues 1-50 of a PVC effector.
16. The PVC Needle Complex according to claim 12, wherein the leader sequence comprises an amino acid sequence having at least 60% sequence identity to one or more sequence selected from SEQ ID NO.: 47-SEQ ID NO.: 92.
17. The PVC Needle Complex according to claim 12, wherein the PVC effector comprises an amino acid sequence of one or more sequence selected from SEQ ID NO.: 1-SEQ ID NO.: 46.
18. The PVC Needle Complex according to claim 12, wherein the PVC effector comprises a sequence selected from SEQ ID NO: 4, SEQ ID NO: 22, SEQ ID NO: 25, SEQ ID NO: 30, SEQ ID NO: 32 and SEQ ID NO: 46.
19. The PVC Needle Complex according to claim 12, wherein the leader sequence is covalently fused to a payload.
20. (canceled)
21. (canceled)
22. (canceled)
23. (canceled)
24. (canceled)
25. (canceled)
26. The effector fusion according to claim 13, wherein the leader sequence comprises amino acid residues 1-50 of a PVC effector
27. The effector fusion according to claim 13, wherein the leader sequence comprises an amino acid sequence having at least 60% sequence identity to one or more sequence selected from SEQ ID NO.: 47-SEQ ID NO.: 92.
28. The effector fusion according to claim 13, wherein the PVC effector comprises an amino acid sequence of one or more sequence selected from SEQ ID NO.: 1-SEQ ID NO.: 46.
29. The effector fusion according to claim 13, wherein the PVC effector comprises a sequence selected from SEQ ID NO: 4, SEQ ID NO: 22, SEQ ID NO: 25, SEQ ID NO: 30, SEQ ID NO: 32 and SEQ ID NO: 46.
30. The effector fusion according to claim 13, wherein the leader sequence is covalently fused to a payload.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0217] Embodiments of the invention will now be described, by way of example only, with reference to the following Figures and Examples.
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EXAMPLES
Materials and Methods
Cloning
[0238] Plasmids encoding PVC Needle Complexes were prepared using standard molecular techniques known in the art. Briefly, genomic DNA from P. asymbiotica.sup.ATCC43949 (obtainable from the ATCC under accession no. ATCC 43949) was used in PCR (with appropriate primers) to amplify multiple (e.g. four) overlapping regions of the PVC operon. Overlap/extension PCR was employed to prepare a whole operon, and fused (again using overlapping PCR) into an appropriate expression vector as detailed in
[0239] Briefly: four overlapping PVC fragments (generated with primers of SEQ ID NO: 101 (F1) and SEQ ID NO: 105 (R1); SEQ ID NO: 102 (F2) and SEQ ID NO: 106 (R2); SEQ ID NO: 103 (F3) and SEQ ID NO: 107 (R3); and SEQ ID NO: 104 (F4) and SEQ ID NO: 108 (R4), respectively) were made covering the PVC operon (e.g. of SEQ ID NO: 93). The target cloning vector was cut at the required insertion site. These 5 DNA fragments were then assembled by overlapping PCR (using primers of SEQ ID NO: 101 and of SEQ ID NO: 108), and the resulting fragment was ligated into the cloning vector. Products were transformed into laboratory E. coli and recovered with vector marker selection (e.g. due to ampicillin resistance).
[0240] The operons are typically operably linked to an inducible promoter (e.g. arabinose inducible, and/or IPTG inducible) as is known in the art. This is generally achieved by cloning into pBAD family plasmids (inducible via arabinose) (Invitrogen, catalog number: V43001) and pVTRa (inducible via IPTG) (Biomedal, S.L.) vectors (although any combination of compatible expression vector systems should suffice).
[0241] A PVC Needle Complex can be expressed independently of the payload (toxin), and vice versa. Separate expression vectors (e.g. having differing inducible promoters) may harbour the PVC Needle Complex and the payload, respectively.
Expression (e.g. laboratory scale expression)/Purification of PVC Needle Complexes in E. coli
[0242] A typical process to purify a PVC Needle Complex from a 1 L culture of an E. coli expression strain (transformed with an appropriate expression vector/cosmid) is as follows: [0243] 1—An overnight culture of the bacteria (transformed with PVC Needle Complex expression vector) is prepared by picking a colony from a plate and inoculating 100 mL of LB media. [0244] The culture is grown at 37° C. with shaking. [0245] a. Typically, the media may be routinely supplemented with 0.2% d-Glucose to aid repression of the genetic constructs for optimal cell health. [0246] b. The media is also supplemented with the relevant antibiotics for maintenance of the expression (PVC Needle Complex) vector. If a payload vector is also being used, the relevant antibiotic for that vector is also supplied. [0247] 2—The next day, a 1 L flask is inoculated via dilution in a 1:100 ratio from the overnight culture. The media for the 1 L flask is identical to the overnight media but typically does not contain glucose. [0248] 3—Cultures are grown to approximately mid-to-late exponential (an OD600 nm of ˜0.8) at which point the plasmids are induced. [0249] a. For the PVC Needle Complex (nanosyringe) plasmid, typically 0.2% arabinose is added to induce expression. For the payload plasmid (plasmid encoding for the payload, such as Pnf), IPTG concentrations may typically be optimised on a per-protein basis, and a typical starting figure of 0.1 mM is preferable. [0250] 4—The cultures are returned to the incubator post-induction and cultured at 18° C. until the following day. [0251] 5—Cultures are harvested by centrifugation in appropriate centrifuges/bottles/rotors at 5000×g for 30 mins. [0252] 6—Cell pellets are then lysed to release PVC Needle Complexes (nanosyringes). [0253] a. The following lysis methods may be used: [0254] (i) Lysozyme incubation overnight. (ii) Sonication with a needle sonicator (with or without first treating with lysozyme. (iii) Cell disruptor/homogenisers. [0255] 7—Optionally, DNAse, and protease inhibitors can be added to the lysate. [0256] 8—Cell debris is removed by centrifugation at 50,000×g, 4° C., for 20 minutes in a high speed centrifuge. [0257] 9—Concentrate the lysate through a 100,000 kDa MWCO centrifugation column to reduce volumes and remove small proteins. Once the volume is down to a manageable volume, centrifuge several times replacing the retentate solution with an appropriate sample buffer such as TM (20 mM Tris-HCl, 8 mM MgCl.sub.2, pH 7.4) to dialyse.
[0258] A subsequent process for purification via Caesium Chloride density gradient is as follows: [0259] 1. Prepare CsCl density solutions as follows: [0260] (a) 1.7 g/mL CsCl in H.sub.2O; (B) 1.5 g/mL CsCl in H.sub.2O; (C) 1.45 g/mL CsCl in H.sub.2O [0261] 2. Gradients (from bottom-to-top of the tube) are then set up in ultracentrifuge tubes like so: [0262] (1) (bottom of tube)—2 mL density, 1.7 CsCl; (2)—3 ml density, 1.5 CsCl; (3)—3 mL density, 1.45 CsCl; (4) (top of tube)—sample in TM buffer. Suitably, apply each density carefully to side of tube so as not to blend the boundary with the previous density layer. [0263] 3. Balanced tubes are then subjected to ultracentrifugation at 35,000 RPM in an SW40Ti swinging bucket rotor, equivalent to 155,000×g, for 2 hours, 4° C. [0264] 4. The correct gradient fraction will be the region just above a ‘blue-ish-white’ halo that appears. Fractions are extracted via puncturing the tube with a syringe and needle. [0265] 5. PVC Needle Complexes of good purity can be obtained in this manner, and stored in buffer at 4° C. Suitably, dialyse back in to TM buffer to remote the CsCl.
[0266] Following, or in place of CsCl gradient purification, PVCs can be extracted via Monolith anion exchange chromatography, as follows (note all steps can be performed manually with a peristaltic pump or syringe apparatus, or via F/HPLC): [0267] 1. Unless already done, dialyse the sample extract into the binding mobile phase (typically TM buffer) with a low concentration of salt (20 mM NaCl). [0268] 2. Equilibrate the column according to the manufacturer's guidelines, briefly: [0269] a. At least 5 Column Volumes (CV) of dH.sub.2O; [0270] b. At least 5 CV of binding buffer (TM, with low salt); [0271] c. At least 5 CV of elution buffer (TM with high salt, >=1M NaCl); [0272] d. At least 10 CV of binding buffer once more. [0273] 3. Apply the sample to the column at a low flow rate (1-2 mL/min) [0274] 4. Wash the column with up to 200 mM NaCl-containing TM buffer. [0275] 5. Elute with 1M NaCl-containing TM buffer (alternatively, use a gradient elution if using an FPLC machine). [0276] 6. PVC Needle Complexes are present in the elution fractions. If a fraction collector is used, subsequent SDS-PAGE or similar may be needed to identify the correct fraction.
[0277] The column (of e.g. step 2) was of the CIMmultus™ Quaternary Amine anion exchange columns (BIA Separations d.o.o.). For example, the CIMmultus™ QA-1, which is a monolithic column with 1.3 μm channel size and a column volume of 1 mL.
[0278] Alternatively, a DEAE (a weak anion exchanger) column may be used.
[0279] Alternatively, for use with a Photorhabdus expression system, PVC Needle Complexes can be purified from supernatants as well as/instead of cell pellets, with the following additions/modifications: [0280] 1. Following cell harvest from the standard protocol above, supernatants are transferred to a pyrex bottle, and can optionally be concentrated via 100,000 MWCO columns if necessary. [0281] a. DNAse (0.25 U/mL) and protease inhibitors can optionally be added. [0282] 2. NaCl is added to a final concentration of 0.5M, and 80 g/L of PEG6000 is also added. The solution is mixed at 4° C. overnight. [0283] 3. The solution is centrifuged to pellet the PEG6000 at 8000×g, 4° C. for 30 mins. [0284] 4. The pellet is resuspended in a small volume (˜5 mL) of TM buffer (or similar) and incubated for 2 hours at room temperature, shaking. [0285] 5. Pellet by centrifugation at 13,000×g for 10 mins, and collect the supernatant to a new tube. Proceed with purification method of choice.
[0286] Other methods for purifying PVC Needle Complexes have been described elsewhere, for example in Yang et al (J Bacteriol. 2006 March; 188(6): 2254-2261), incorporated herein by reference.
Construction of an arabinose inducible over-expression strains for P. luminescens TT01 PVCunit4 (chassis encoded by genes Plu1667-plu1652)
[0287] Photorhabdus strains overexpressing a PVC Needle Complex were prepared using chromosomal recombineering to place a PVC (operon) of choice (operon encoding PVCunit4 Needle Complex was used here, as an example) under the control of an arabinose inducible transcription promoter. The recombineered strains are then genetically transformed with effector expression plasmids (e.g. based on the arabinose inducible expression vector pBAD30) to facilitate PVC Needle Complex over-expression, PVC effector expression, PVC effector trans-packaging, and secretion of the whole complex simply through the addition of the arabinose sugar.
Recombinant Photorhabdus PVC over-expression strain construction
[0288] The promoter region of PVCunit4 was amplified using primers PVCpromF (5′-TATCATATGTCTACAACTCCAGAACAAATTGCTG-3′, SEQ ID NO: 97) and PVCpromR (5′-ATCTCTAGAACAGATATTCCAGCCAGC-3′, SEQ ID NO: 98) using genomic DNA from P. luminescens strain DJC (aka strain TT01) as a template. A suitable P. luminescens strain is obtainable from the ATCC under accession no. ATCC 29999. The PCR product was digested with NdeI and XbaI and introduced by ligation into the suicide vector pCEP (ThermoFisher, catalog number: V04450), using E. coli DH5α λ-pir (Biomedal S.L.) as the carrier strain. The resulting plasmid was transferred to the E. coli donor strain S17.1 λ-pir (Biomedal S.L.) for conjugation into Photorhabdus. Briefly, overnight cultures of the donor strain and a rifampicin resistant (RifR) isolate of P. luminescens DJC were diluted in LB supplemented with 10 mM MgSO.sub.4 and grown to mid-exponential (OD600 ˜0.5). Then, 3 ml of each culture were harvested, washed twice and re-suspended in 100 μl of LB supplemented with 10 mM MgSO.sub.4. 80 μl of P. luminescens DJC RifR were mixed with 20 μl of the donor bacteria (resulting in a recipient to donor ratio of 4:1) and placed in the centre of an LB agar plate supplemented with 0.1% pyruvate and 10 mM MgSO.sub.4. The plate was incubated overnight at 30° C. and the resulting growth was harvested in 1.5 ml LB. Aliquots were plated on plates containing rifampicin (50 μg/ml) and chloramphenicol (25 μg/ml) to select for trans-conjugants and the plates were incubated at 30° C. for 3 days. Possible transconjugants were re-streaked and confirmed by PCR using primers ParaINF (5′-GGCGTCACACTTTGCTATG-3′, SEQ ID NO: 99) and tPVCpR (5′-TCGGTGGCAGTAAATTGTCC-3′, SEQ ID NO: 100).
PVC Needle Complex over-expression and purification from Photorhabdus
[0289] Overnight cultures of P. luminescens DJC PVCunit4::pCEP were diluted in 2×250 ml LB supplemented with chloramphenicol (25 μg/ml) and incubated at 28° C., 180 rpm. After 2-3 h, arabinose (0.2%) was added and the cultures were returned to the incubator for another 26 h. The cells were pelleted by centrifugation (7000 g for 30 min) and the supernatant was collected. DNAse I was added to the supernatant at a concentration of 0.25 U/ml to degrade any extracellular DNA. Following an incubation of 30 min at room temperature, polyethylene glycol 8000 (8%) and NaCl (0.5 M) were added to precipitate the proteins. The supernatants were incubated overnight at 4° C., stirring. The precipitated proteins were then collected by centrifugation at 8000 g for 30 min at 4° C. The pellets were re-suspended in 8 ml TM buffer (20 mM TrisHCl, 20 mM MgCl2, pH7.4) and incubated at room temperature for 2 h with gentle shaking. Any remaining debris was removed by centrifugation at 13000 g for 10 min and the supernatant containing PVC Needle Complexes was applied to a CsCl density gradient and centrifuged at 35000 rpm for 2 h in a Beckman coulter Optima L-90K or XPN-80K ultracentrifuge. The CsCl density gradient was made by layering TM buffer containing CsCl at p=1.7 (2 ml), 1.5 (3 ml), and 1.45 (3 ml) from the bottom of the tube, respectively. The fraction containing PVC Needle Complexes was collected and Ultracel-100K devices (Amicon) were used to remove the CsCl and exchange the buffer for TMS (20 mM TrisHCl, 8 mM MgSO4, pH7.4). The PVC Needle Complexes were further purified using a CIMmultus™ quarternary amine 2 μm pore anion exchange column (BIAseparations). The column was washed with TMS buffer containing 200 mM NaCl and the PVC Needle Complexes were eluted in TMS containing 1 M NaCl. The NaCl was removed by buffer exchange using an Ultracel-100K device and the sample was applied to a CIMmultus™ DEAE 2 μm pore column (BIA separations) for a final purification. The column was washed in TMS containing 200 mM NaCl and the sample was eluted in TMS containing 500 mM NaCl.
[0290] It is possible to perform this with and without lysis (e.g. because the PVC Needle Complexes appear to be secreted from live cell, and can be collected in supernatant) of the cells (to release the PVC Needle Complexes).
Transmission Electron Microscopy
[0291] For transmission electron microscopy (TEM) pioloform-covered 300-mesh copper grids that were coated with a fine layer of carbon were used as substrates for the protein fractions. A preferred aqueous negative stain is 3% methylamine tungstate. The coated grids were exposed to UV light for 16 h immediately prior to use to ensure adequate wetting of the substrate. A 10 μl drop was applied to the TEM grid, and the protein was allowed to settle for 5 min. Liquid was absorbed with filter paper from the edge of the grid and replaced immediately with 10 μl of filtered negative stain. The drop was partially removed with filter paper, and the grids were allowed to air dry thoroughly before they were viewed with a JEOL 1200EX transmission electron microscope (JEOL, Tokyo, Japan) operating at 80 kV.
BioPORTER assay and actin stress fibre analysis.
[0292] For BioPORTER assays (Genlantis), 80 μl of purified wild-type and mutant Pnf proteins (500 μg ml-1), or PBS as a negative control, were added to one BioPORTER tube (Genlantis) and re-suspended in 920 μl of DMEM. The samples were added to HeLa cells grown in 6-well plates and incubated for 4 h. BioPORTER/protein or PBS mixes were replaced by fresh complete medium and the cells were incubated for 20-48 h. To visualize cell morphology and actin cytoskeleton, cells were fixed for 15 min in 4% PBS-formaldehyde, permeabilized with 0.1% Triton X-100 and stained with Tetramethylrhodamine B isothiocyanate (TRITC)-phalloidin (Sigma) and DAPI dihydrochloride (Sigma). Images were acquired with a LSM510 confocal microscope (Leica).
Example 1
Cloning and Expression of PVC Needle Complexes
[0293] The inventors have successfully excised (cloned) the required expression genes from the host bacterium, Photorhabdus (e.g. which are comprised within SEQ ID NO: 93, SEQ ID NO.:94 and/or SEQ ID NO:95), and have devised a reliable, scalable expression system in laboratory E. coli as explained above. It has been demonstrated that trans-expression on separate plasmids enables incorporation of payloads (e.g. Pnf) into the syringes, creating a multi-plasmid (modular) platform.
[0294] Following purification from E. coli, electron microscopy analysis demonstrated that the purified PVC Needle Complexes retained the correct ‘nanosyringe’ structure (see
[0295] Furthermore, electron microscopy analysis demonstrated that the purified complexes appropriately localise to the cell surface of cells, and PVC Needle Complexes with a Pnf payload (PVCpnt) induces a phenotype (ruffling) consistent with the postulated mechanism of the effector (PVC)—see
Example 2
2.1 Demonstrating PVC Needle Complexes Exert Effect Via Intracellular Delivery of Effector
[0296] The polypeptide Pnf was identified as a PVC effector as follows. This was identified within the Photorhabdus asymbiotica ATCC43949 complete genome—GenBank Accession Number: FM162591.1.
[0297] The final gene of the PVC operon (P. asymbiotica ATCC43949 PVCpnf operon, which has a sequence of SEQ ID NO: 93) was identified, namely pvc16 (e.g. PAU_03338). The position of the pvc16 genes of a PVC locus is illustrated in
[0298] A Pnf loaded PVC Needle Complex was then prepared according to Example 1.
[0299] The inventors have demonstrated that these packaged (e.g. laden) PVC Needle Complexes exert cellular effects consistent with the provenance of the cargoes they carry. By way of example, cells and whole insect animals exposed to PVC Needle Complexes loaded with the cytoskeleton toxin Pnf undergo cell death in a manner consistent with cytoskeleton toxicity.
[0300] Injection experiments (injection into the insect larvae) were performed by injection of 10 μl of supernatant, provided following centrifugation (pelleting) of an overnight culture (typically 1 L) of a culture of E. coli harbouring a cosmid clone encoding the PVC Needle Complex with Pnf (PVCPnf)—e.g. a PVC encoded by SEQ ID NO.: 93, packaged with a PVC effector of SEQ ID NO.: 32.
[0301] Demonstrating that the PVC Needle Complexes are responsible for the phenotype due to intracellular delivery (e.g. injection) of the Pnf payload, the toxic effect could only be reconstituted when the same protein (Pnf) is provided with another route to access the cell cytosol (transfection and expression of an expression plasmid, or conductance via liposomal preparations containing the protein)—see
2.2. Evidence of Delivery of the Toxic Effector Enzyme Pnf into Cultured Human Macrophages
[0302] To complement the data outlined above, the inventors conducted additional experimentation providing further evidence of delivery of the toxic effector enzyme Pnf into cultured human macrophages.
[0303] Concept: The inventors tested PVCpnf expressed and purified from E. coli, (trans-)packaged with the native Pnf toxin on cultured human THP1 derived macrophages. Unlike the lethal effect of the Pnf toxin in insect models, previous liposome mediated Pnf protein transfection experiments indicated a subtler phenotype in human Hela cells. In those experiments the cells showed actin stress fibre formation at 24 h and multinucleation at 48 h. The inventors therefore tested the effect of the purified PVCpnf (the nanosyringe) holding/packaged with the Pnf PVC effector on macrophage respiration rate using a Resazurin colourimetric assay.
Methods:
[0304] Background behind Resazurin assays. The blue compound resazurin was explored for use in assays to determine the activity of PVCs on macrophages (MO). Resazurin is metabolically reduced in cell mitochondria, producing a pink and highly fluorescent compound, resorufin. The effect of PVCs on macrophage metabolism can be determined by introducing resazurin into the culture media. The number of macrophages affected by PVCs can be inferred by comparing the fluorescence measured to that of the cell density optimisation curve (see Czekanska, Methods in Molecular Biology, 2011, 740, 27-32, incorporated herein by reference).
[0305] Optimisation of use of Resazurin for THP1 derived macrophages. The metabolism of macrophages over 18 h was assessed at different seed densities to determine the optimum cell density for use of this assay with PVCs. A 30 mL culture of THP-1 cells was pelleted at 1000 rpm for 4 min, before resuspension in 2 mL of RPMI media (also containing 10% FBS (v/v) and 2 mM L-glutamine). Cells were counted using a cell haemocytometer, then diluted in media to a density of 2×10.sup.6 cells mL.sup.−1. THP-1 cells were then activated with phorbol 12-myristate-13-acetate (PMA) immediately before plating. 200 μL of the cells were plated in quadruplicate in a 96-well plate, and a 2-fold serial dilution was performed until reaching a final cell density of 1.5625×10.sup.3 cell mL.sup.−1. 125 μL of the starting cell dilution was also plated in quadruplicate on the same plate, for a 5-fold serial dilution, until reaching a cell density of 0.32×10.sup.3 cells mL.sup.−1. Four blank wells were also prepared, containing RPMI and PMA. The plate was incubated at 37° C. with 5% CO.sub.2 for 48 h. Media was aspirated from the wells and replaced with fresh RPMI, and the macrophages were incubated for a further 24 h. A resazurin tablet (VWR) was dissolved in RPMI (12.5 mg/mL), and 10 μL added to each well in quick succession (well concentration of 1.25 mg/mL). The fluorescence produced was measured on a plate reader every 30 min for 18 h (excitation: 530-570 nm, emission: 580-620 nm, maintained at 37° C. and 5% CO.sub.2). The optimum cell density over time was then determined for use with PVCs.
[0306] Use of assay for PVC testing. THP-1 cells, diluted to 1.25×10.sup.5 mL.sup.−1, were activated and seeded in a 96-well plate, where wells contained 100 μL of cells at a final well density of 1.25×10.sup.4 cells mL.sup.−1. Blank wells were also prepared in quadruplicate, containing cells without PVC samples, as well as wells containing media and PMA only. The plate was incubated for 48 h at 37° C. with 5% CO.sub.2. The media was then replaced with fresh RPMI, before addition of 10 μL of each PVC sample. The plate was incubated for a further 24 h, before the addition of 10 μL resazurin (12.5 mg/mL) to each well, and the fluorescence was measured every 30 min for 18 h (excitation: 530-570 nm, emission: 580-620 nm, maintained at 37° C. and 5% CO.sub.2).
[0307] Results:
Example 3
[0308] Demonstrating that a Leader Sequence is Responsible for Payload Packaging into PVC Needle Complexes
[0309] Surprisingly, the inventors have found that the provision of a ‘leader’ peptide sequence, preferably on the N-terminus of a payload (toxin) protein, can direct the payload to the PVC complex and allow for (e.g. trigger) the packaging of the payload into the PVC Needle Complex. The inventors have demonstrated that amino acid residues 1-50 of a PVC effector protein is/comprises a leader sequence.
[0310] To demonstrate this, an expression construct (overexpression in chromosomally engineered P. luminescens TT01) was prepared, in which the leader sequence (the N-terminal amino acid residues 1-50) was ablated such that the payload expressed by Plu1649 (referred to as “hvnA” in the figure, and having a sequence of SEQ ID NO.: 46) (Myc-tagged for detection purposes) was absent a leader sequence (see
[0311] Surprisingly, hvnA having a leader sequence from a different (non-hvnA) PVC effector (i.e. corresponding to the N-terminal amino acid residues 1-50 from the PAU_03332 effector) (see
Example 4
[0312] 4.1 Demonstrating that a Leader Sequence Directs Packaging (into PVC Needle Complexes) of Atypical/Exogenous Payloads
[0313] In an unexpected technical effect of the invention, the inventors have found that fusing a leader sequence described herein to exogenous (non-Photorhabdus) polypeptides (preferably at the N-terminus) allows for packaging of said exogenous polypeptides into a PVC Needle Complex, with the exogenous polypeptides remaining associated with the PVC Needle Complex upon isolation/purification. By way of example, see
[0314] The inventors performed in-depth analysis of the size (e.g. polypeptide length) and structure of the various natural PVC effector payloads encoded by Photorhabdus (see
[0315] Furthermore, this packaging of exogenous polypeptides is independent of the chosen PVC Needle Complex chassis e.g. has been accomplished using both a “PVCpnf” chassis (SEQ ID NO.: 93) and a “PVCU4” (e.g. PVCunit4) chassis (endogenous to the Photorhabdus overexpression strain) (see
[0316] In data shown herein, payload proteins are supplied in ‘trans’ on separate genetic constructs. The leader sequences are surprisingly sufficient to target these separately synthesised proteins for packaging into the PVC Needle Complex vehicle (see
[0317] Further exemplification of trans-packaging of high levels of the Cre site specific recombinase into the PVCpnf nanosyringe expressed in E. coli is provided in
4.2 Trans-packaging using additional leaders demonstrating functionality of a larger, diverse sequence space
[0318] Complementing the data outlined in Example 3,
[0323] These results also demonstrate the utility of leader sequences showing greater sequence diversity for (trans-)packaging a payload. Indeed, to provide further validation, the inventors performed a CLUSTALW sequence comparison of a panel of leader sequences to determine diversity. PVC effectors are identified as proteins encoding recognisable toxin-like domains that are encoded immediately downstream of the pvc16 structural gene. Each PVC operon can encode just a single effector, or several different effector genes in tandem array. A phylogenetic tree is shown in
[0324] As can be seen from the tree of
Example 5
Tail Fibre/Binding Domain Modification
[0325] PVC Needle Complexes are known to comprise tail fibres (see the 3D rendered PVC structure, left most asterix of the rightmost image) which are believed to allow for cell-type specific targeting of the PVC complexes. The inventors have successfully demonstrated that modification of a tail fibre region to incorporate non-natural amino acids (e.g. a substitution of an amino acid in the wild-type sequence for an alternative amino acid of the 20 standard amino acids) does not affect expression of tail fibres.
Example 6
[0326] Demonstrating Delivery of an Active (Exogenous) Enzyme/Payload into Ex Vivo Murine Organoids with a Leader Sequence-Packaged PVC Needle Complex
[0327] Concept: Obtaining data for the delivery of an exogenous functional enzyme to a mammalian tissue. The inventors have demonstrated the delivery of a trans-packaged bacteriophage derived recombinase protein known as “Cre” into ex vivo mouse bile duct organoids. The organoids are derived from a mouse line in which the expression of a chromosomally encoded red fluorescent protein (RFP) reporter is normally prevented by a stop signal flanked by loxP recognition sites for the Cre-recombinase. If the recombinase is present, the stop signal is recombined out and the cells then go on to express the reporter protein. The general principle behind this experimental demonstration is summarised in
[0328] Method: The Bile Duct organoid preparation: murine primary bile ducts were isolated and expanded as organoids in matrigel using “BD expansion media” for 12 passages following Huch et al (Regen Med. 2013 Jul.; 8(4):385-7. PMID: 23826690; DOI:10.2217/rme.13.39) protocol. Cells were then plated in 2D and cultured in BD expansion media. Mouse Genotype: LSL-Tom reporter in Rosa26 locus+Axin2CreRT (inducible upon 40HT treatment). Cells were cultured in uncoated polystyrene plates at a seeding density: of 10,000 cells/well. Nanosyringes were prepared as 30% volume syringe preparation in PBS+70% culture media. Total volume of 100 μl per well. The positive control represented 500 nM 40HT (in ethanol) at 1:1000 (v/v) as positive control for the recombination. The negative control represents 1:1000 (v/v) ethanol dilution only. Cells were seeded and grown for 48 h, nanosyringes added and then cultured for another 24 h before fixing (4% PFA fixation 15 min RT) and staining for microscopic examination. Staining: Primary antibody Anti-RFP (1:1000) from Rockland. Secondary Anti-Rabbit 568 (used at 1:500 v/v). Samples were visualized on a laser-confocal microscope.
[0329] Result:
[0330] Additional interpretation: To summarise, the inventors have demonstrated the ability to deliver (e.g. dose) exogenous enzymes to a cellular target. Moreover, this “nanosyringe+Cre” experiment is a promising proof of concept for a biotechnology tool/aide, by demonstrating the ability to provide a DNA change leading to a transformed cell. This experiment therefore demonstrates the use of exogenous payloads (a protein of viruses rather than bacteria), and nucleic acid modifying enzymes in particular. It is evident that the Cre enzyme is delivered in a functional manner and is capable of traversing the cellular interior to the nucleus to affect its DNA modifying changes.
Example 7
[0331] Trans-Packaging of MAD7 Site Specific Recombinase (Exogenous Payload) into the PVCpnf Nanosyringe Expressed in E. coli
[0332] Concept: As with the Cre data (of Example 6), and other examples of packaged payloads provided herein, the inventors have demonstrated packaging of the Cas-like enzyme MAD7 into a nanosyringe via a leader sequence. This is the largest exogenous example (MAD7=147.9 kDa) of a payload described herein.
[0333] Methods: Briefly, the chassis genes and the MAD7 gene (the latter being tagged with a C-terminal Myc tag for detection, and a leader sequence for nanosyringe incorporation described herein), were expressed (upon induction) simultaneously in E. coli. Upon harvesting and purification of the nanosyringe complex, payload packaging was probed via dot blot analysis (e.g. for detection of the Myc tag). The purification method described herein (using ultracentrifugation) can be employed to select for (e.g. exceedingly) high molecular weight protein complexes/biological matter, enabling recovery of the nanosyringes and any cargo (payload) they carry. ‘Loose’/unpackaged payload remains in solution and is not subject to sufficient centrifugal force and as such is lost during purification, unless contained within the much larger nanosyringe ‘shell’ (that is, when successfully packaged). Successful packaging of MAD7 is demonstrated by
Example 8
[0334] Trans-Packaging of Apoptosis Inducing Payloads into PVCpnf, Expressed in E. coli
[0335] Using the E. coli PVCpnf leader::payload::Myc trans-packaging system described in
Example 9
[0336] Demonstration of the induction of apoptosis in cultured ex vivo human cells by nanosyringe delivery of (trans-)packaged pro-apoptotic human polypeptides
[0337] A preliminary test has confirmed the ability to use the PVCpnf nanosyringe, produced in E. coli, to deliver trans-packaged human protein sequences (e.g. packaged according to Example 8) and induce apoptosis in ex vivo circulating PBMC cells from human donors. The assay is a TUNEL-stain microscopic analysis from cells exposed to the packaged nanosyringes for 20 minutes only. Results are shown in
[0340] A more detailed test of the delivery of pro-apoptotic human peptides into ex vivo Peripheral Blood Mononuclear Cells (PBMCs) is now described. The aim of this study was to investigate whether the pro-apoptotic peptide loaded PVC nanosyringes could induce apoptosis in ex vivo human Peripheral Blood Mononuclear Cells. The nanosyringes were first assessed for any immediate cell toxicity using Trypan blue dye exclusion assays and then for apoptosis response by using the TUNEL assay.
[0341] Trypan Blue Exclusion Test for cell viability: Trypan blue is a Diazo dye commonly used to selectively colour dead tissue or cells, hence, dead cells are shown as a distinctive blue colour under a microscope while live cells or tissues with intact cell membranes remain uncoloured. Since live cells are excluded from staining, this staining method is also described as a Dye Exclusion Method. Trypan blue is commonly used for assessment of tissue or cell viability. A suitable number of cells (2×10.sup.5) were exposed to the nanosyringes and empty nanosyringe for 20 minutes. A suitable volume of cells (30 μL) were added to an equal volume of 0.4% Trypan blue and the number of viable (unstained) and dead (stained) cells counted using a hemocytometer. Each compound was tested at 3 concentrations. Blood cells from two independent human donors was tested for each compound at each concentration and each sample was tested in duplicate.
[0342] Treatment and preparation of cells for microscopy: The viability of Peripheral Blood Mononuclear Cells (PBMCs) from two independent healthy human donors was determined after 20-minute treatment with the two chimeric nanosyringes (e.g. loaded with the exogenous pro-apoptotic peptides) at 3 test concentrations in 2 independent tests. PBMCs were harvested by centrifugation and resuspended in media at 1×10.sup.6 cells/ml. Cells were fixed in 2.5% formalin and incubated for 20 mins at room temperature. Poly-L-lysine coated slides were prepared by spraying with 70% ethanol and leaving to air dry. Cells were centrifuged for 30 seconds. Supernatant was removed and cells were resuspended in 200 μl dH.sub.2O. 5 μl of cell suspension was added to each slide/fixation. Two fixations were performed per slide to allow staining to be performed in duplicate. Cell suspension was left to air dry.
[0343] Results of PBMC cell viability assay: The Trypan blue viability assays confirmed that the PVC preparations were not immediately toxic in themselves to PBMCs taken from healthy human donors (Table 2). Nanosyringe treatment showed >60% viability indicating low toxicity at maximum dose concentration (Table 2). The inventors then moved on to test the ability of the chimeric nanosyringes to induce apoptosis.
TABLE-US-00003 TABLE 2 Viability of Peripheral Blood Mononuclear Cells (PBMCs) from two independent human blood donors after exposure to each compound for 20 minutes at 3 test concentrations (v/v dilutions). PBMC controls are untreated. Neat 1/10 1/100 Well 1 Well 2 Well 1 Well 2 Well 1 Well 2 Blood donor sample 1 TBID 60 70 75 75 80 80 Bax 78 82 80 80 83 87 PBMC Controls 93 97 Blood donor sample 2 TBID 70 72 81 81 84 84 Bax 80 80 84 88 87 87 PBMC Controls 95 95
[0344] Testing for chimeric nanosyringe induced apoptosis using the TUNEL assay: The TUNEL assay was then used to identify apoptotic nuclei in single cell suspensions fixed on slides. In the assay Terminal deoxynucleotidyl Transferase (TdT) binds to the exposed 3′-OH ends of DNA fragments which are generated in response to apoptotic signal factors. This in turn catalyses the addition of biotin-labelled deoxynucleotides which can be detected using a streptavidin-horseradish peroxide (HRP) conjugate. Diamineobenzidine (DAB) reacts with the HRP-labelled sample to generate an insoluble brown substrate at the site of DNA fragmentation. Methyl green counterstaining enables the visualisation of normal and apoptotic cells.
[0345] The induction of apoptosis following exposure of human PMBCs to the nanosyringes was determined. A TUNEL assay kit (Abcam) was used for detection of apoptotic cells. The assay was performed following the manufacturer's instructions. Briefly, slides were covered with 100 μL proteinase K solution or 5 minutes, slides were rinsed with 1×TRIS buffer saline (TBS). The treatment of nanosyringes or the DNase I positive kit control was performed for 20 minutes at room temperature. Slides were rinsed with TBS. Slides were then incubated with TdT equilibrium buffer for 30 minutes before the addition of TdT labelling reaction mix. Slides were incubated at 37° for 19 minutes. Slides were then washed with TBS before application of the stop buffer and incubation at room temperature for 5 minutes. Slides were washed again with TBS before addition of the blocking buffer for 10 minutes at room temperature. Detection was performed by application of the conjugate to the samples for 30 minutes. Slides were rinsed with TBS before application of the DAB solution for 15 minutes. Slides were rinsed with dH.sub.2O followed by counterstaining with methyl green. Slides were dehydrated in 100% ethanol followed by xylene and mounted with a glass cover slip. All staining was performed in duplicate. An apoptosis endpoint, indicative of positive staining in the apoptosis detection assay is represented by dark brown (DAB) signal. Lighter shades of brown and/or shades of blue/green to green/brown indicate a non-reactive negative cell for apoptosis.
[0346] Analysis was performed by selecting 5 random sections of cells on the slide, positive stained cells (dark drown) and negative stain cells (blue or light brown) were counted and the percentage of cells showing apoptotic bodies was determined.
[0347] To generate a positive control, slides were treated with 1 μg/μl DNase I (the kit positive control) for 20 minutes at room temperature following the proteinase K treatment step detailed below. The DNase I treatment fragments DNA in normal cells to generate free 3′OH groups identical to those generated during apoptosis. A negative control was generated by substituting DNase I with dH.sub.2O in the reaction mix during the treatment stage.
[0348] Results of PMBC apoptosis assays: TUNEL staining using PBMCs was performed following treatment with the intact tBID and Bax loaded nanosyringes, with appropriate positive and negative kit controls. Treatment was performed for 20 mins to determine if the nanosyringes elicited an apoptotic signal. A positive control (DNase I treatment) and negative control (no DNase I treatment) was included. Results showed both nanosyringes, containing either tBID or Bax, showed strong apoptotic signals (89% and 78% positive, respectively) on the PBMCs. The positive control showed a strong apoptotic signal (79%), whereas the negative control showed no apoptotic signal (100% negative). Also observed was a significant loss of the numbers of attached cells in the nanosyringe treated samples, presumably indicative of a rapid and comprehensive apoptosis response, and a failure to be retained after washing. Note this effect is even more pronounced than the kit positive control suggesting a more rapid response. Representative micrographs are shown in
[0349] Conclusion: It is concluded that the tBID and Bax loaded nanosyringes are able to rapidly induce extensive apoptosis in human Peripheral Blood Mononuclear Cells. Furthermore, Trypan Blue dye exclusion assays have confirmed that these chimeric nanosyringes do not cause rapid lethal lysis or extensive membrane damage to the cells.
Example 10
Exemplification of Practical Utility of Leader Sequences and PVC Needle Complexes—Intracellular Delivery of Atypical (Non-Photorhabdus) Payload
[0350] (1) An anti-MDM (p53 inhibitor) antibody is linked to a leader sequence described herein, and expressed together with a PVC Needle Complex for packaging therein. Isolated PVC Needle Complex (comprising the antibody payload) is contacted with a tumour for intracellular delivery of the antibody (said tumour cells being characterised by having high MDM-suppression of p53 activity for MDM inhibition). The tumour is suppressed by the activity of the anti-MDM antibody.
[0351] (2) A PVC Needle Complex is used to (intracellularly) deliver anti-tumour peptide vaccine to activate the MHC-I dependent cytotoxic T-cell lymphocyte (CTL) response. A tyrosinase-related protein 2 (TRP2) peptide vaccine is delivered for enhancing cross-presentation to CTLs occurs and antitumor effects against TRP2-expressing tumours. The tumour is suppressed by the activity of the peptide vaccine.
[0352] (3) A PVC Needle Complex is used to (intracellularly) deliver a nuclear factor-KB inhibitors (which are used for the control of inflammatory disorders, such as rheumatoid arthritis) to a cell. The cell subsequently demonstrates a reduced expression of pro-inflammatory cytokines.
[0353] (4) A PVC Needle Complex is used to (intracellularly) deliver a T3SS payload (which inhibits NF-kB and MAPK pathways). This is completed with an isolated (purified) PVC Needle Complex, without any need for the PVC Needle Complex to remain associated with the bacterial cell from which it derives.
[0354] (5) A PVC Needle Complex is used to (intracellularly) deliver, to a cell, anti-apoptotic peptides including BH4, the Bcl-xL-protein, and/or a peptide inhibitor of c-Jun N-terminal kinase (which can protect the heart and brain against ischemic injuries (a restriction in blood supply to tissues, causing a shortage of oxygen and glucose needed for cellular metabolism)). For example, Jun-kinase inhibition via a 20 amino-acid binding motif of the JUN kinase is sufficient. A release of e.g. cytochrome c in the cell is inhibited.
[0355] (6) A PVC Needle Complex is used to (intracellularly) deliver nicotinamide adenine dinucleotide quinone internal oxidoreductase (Ndi1), the single-subunit yeast analog of complex I (which provides significant cardioprotective effects) to complex I-deficient mutant cells. The Ndi1 protein is correctly targeted to the matrix side of the inner mitochondrial membranes, and restores the NADH oxidase activity to the complex I-deficient cells.
[0356] (7) A PVC Needle Complex is used to deliver one of two of the essential subunits of the PHOX complex (which are used in enzyme replacement therapy to restore production of ROS in chronic granulomatous disease) to a chronic granulomatous disease cell. A restoration in production of ROS is observed.
[0357] (8) A PVC Needle Complex is used to (intracellularly) deliver (e.g. intramuscularly) a myotubularin (which is used for improving local and distant muscle performance in X-linked myotubular myopathy patients). Myotubularin—dephosphorylation of phosphatidylinositol 3-phosphate and phosphatidylinositol (3,5)-bi-phosphate is observed.
[0358] (9) A PVC Needle Complex is used to (intracellularly) deliver a recombinase “Cre” (which is capable of excising defined genetic cassettes) into a mouse cell line, in which the genome has loxP recombination sites flanking a stop signal upstream of an mCherry gene. The Cre payload excises the recombination sites, and removes the stop signal, allowing for expression of the mCherry gene in the cell.
[0359] (10) A PVC Needle Complex is used to (intracellularly) deliver a ˜15 kDa nanobody (antibody fragment) with affinity for an intracellular component. A nanobody-intracellular complex is detected.
[0360] (11) A PVC Needle Complex is used to intracellularly deliver (e.g. into insect cells) an atypical (non-Photorhabdus) polypeptide toxin for insect crop pests and animal parasites. Suppression of the pests is observed.
[0361] (12) A PVC Needle Complex is used to (intracellularly) deliver a nuclease (e.g. Cas9 and/or Mad7) into a target cell comprising a guide RNA. The nuclease performs site-directed gene inactivation
[0362] All publications mentioned in the above specification are herein incorporated by reference.
[0363] Various modifications and variations of the described methods and system of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in biochemistry and biotechnology or related fields are intended to be within the scope of the following claims.
TABLE-US-00004 SEQUENCES Where an initial Met amino acid residue or a corresponding initial codon is indicated in any of the following SEQ ID NOs, said residue/codon may be optional. (PAK 1985) SEQ ID NO: 1 MMREYSNEDDFIKEKTNLVKSENVEADNYLETEYLTYLAKLIGMTERENHHLNSIKLIDDIIELHNDRKGNKLL WNDNWQDKIIDRDLQSIFKKIDEMVSEFGGLEAYKDIVGENPYDPTEPVCGYSAQNIFKLMTEGEYAVDPVK MAKTGKINGNQFAEKLEHLNSSNNYVALINDHRLGHMFLVDIPSTNRERVGYIYQSDLGDGALPALKIADWL KSRGKESINVNKLKKFLNDEFTMLPDNEQKGLIAEIFDLNKDIDSVKSGKIKKDKAVDIYLREYDINDFISNIEKL KTKLA (PAK 1987) SEQ ID NO: 2 MFQNRIRNEKTTQSGKGKTLDRMTDSLYLEIPNVEAVTLAYQKLTSKYRKFDNKTKLILDSSDEFSQLKSEK QRKGFSKSGLKNNGVSDRKFIYTKNALKNFAAHAGYEHNGHYEDEFVNFKDNNKNLAKGKLFPGISLIERR KLSIVKNKEGKWEHKETDEAEAYKVTDIEKFISGVRSMYLQGNTFLHAKTEALIRKHIANNENILPTMAGIAGL HAEVQALNNLFISGDKGTKKREKWKYIRNMLESSIFTQRLTTGQAGKDFAACHNCSGILSSPVNVITGKVES AGDNFLSTLSRYKTSQESPI (PAK 1988) SEQ ID NO: 3 MEREYSEKQKNPSKLSRKTAISERIAALERSGLSNSNQPVPQFARPYTSNRPVVNINPGRSSIAVATANSTS PVNIPTPAPASPDKLLPSTSCDTTSSILIVGKYNLELTSQGKIVVFRGDNRTPEQIVAAGGFYPWSKQDVGKI KKELIDEFIEIGPSAHMMGHVRSPNKNYVSTGMNMDSGGFGEQSNYLYKMEIPGLKPQDMNERTLGEKIRQ DKRGINYPHFLMSHLTLAESEFVAMIPARSEELTFITPIPLSYITSYRKRGTNTWLPMPLKK (PAK 2075) SEQ ID NO: 4 MSNYEYDIVTQHDTYQIKDNEYTVVNGKYWQYEQEGNKNNNKVSISLMKENQNDPVWITSDIKEISLYIIENL FSYHKFSAELQHTLKNAVKAVFNEYSEIKYSELLHNINNIFNLFFIKIYNTSDIDTAINILTAKIEIYDKLEKINQDK TDSNNTNVDIWEELGINAEEPLLKIYRQAFSTGDIDDEVYSDALLTFMSDGNLELGDKEKSDYNQRIKDKTDL FESYKKGIEKVASLITTNNINPGIPITYPETEKSINIGDDLLLAQLAKEEIALKKQNRTEYSQQDIFELQTLQAAK YHLLILSSLGALLYQIAPNVEKMTKGHGDYRDIIFSQEQAESLFKKHNIQYDTNHVLSQESKHIEMEGCIILTAA IIYRMRKENATVEQALNYSTLETIKLFENDKKKLNPFNTNNVKPAGYFSFIDFKKRDKFDSQYNFNEQFNVYK NKYSHYESISFSKLILSSPAAQLTAEEIVNPPEEAFLYSVEQGMGNVAMIKMYQGNWLVISTIQGGVKAKKYS RQQVDSNPTLRAMSKPNALFLIERKMETGMGILMPNMMVNTGKRLFPTGYERAKTLSGFAETSRYKNSYN AFWNDYYGITSGMNVGISFTGSPKFNFYKEENLLSVTATIIQQGLNDIAIKSKQALDITSGWHIAATILIPFYNVI YKSTTDSEYELTGEDIGSIVFDTANVLLVVATLGMSLTESMAAKVTQTTLRLRQAGLTGRALITAVVRTLPEH GIITLRQSSGIILGGLIDLIEPLPIRSTLTLTYRGVINAVGAMRNSIKLEKSFADIFGKSTRGLGKLKNEWKVSNL PLEEIVPHSNGGEIYKGIYSIRPTNPETAVKQNFYIKEAGANYQVKWDDANHTWRVVNPTYPEQFSYWPAV KLDKNGHVWTHADVSNKFLILEQSKRIDQELEAAHSNINNDNILDAFIHINTAFKDCERYDIDKLSDITDTLTHF FEKSLKPGDKKAIFSTEIMSIQQAWIREVILPLQNNSSISIEKINAIKTELPYLLRKTFPIESQLPNQLVANKIALAI EEIPNTRIPKYTSGNISKTVQYTSLLENNHVDIPPVGITITGNDTFINQVTRVLSEIDEIPSGNIVIQELEKQGLNI QPPTMNDIVREKNGQFYANNSAGSHIAFDPENHLIGTEEKLIDEPWRTREPAIALYHEMLHIYYNRYPTWFT SIDNKVIDQKVSGGFSLLEESRIVGTKYYVNDKNTLFDFNDSDYLLENNSALLTENRFRAEYAIFKNKSEYVIR PYSGKGDSQIPLTKTKININESHRNVMGVGSGKPEKMPNESATDYRNRVREWRKANKQPEADIGTGDMRK TKAEARVKLLKENYPQFEPQKIELGGAFQLWTVPNEPANKLMLSSHGYFFSDSAATQVPAGKTIQFLGPHG KTLLEAPENPLYSPFDVTLGNSGFTVQPYATIESGNKAGLGSVKIGDKTFTVNDIQNIATDDVENYLLATGVE ANASNHGKVRNYGIKYYEKMPDEEVKAAIWKNRADETSTHKYDALLVSPEAGNRKKLSDIFALMKTDERMS KYDEITFVACREELNRINMKSIHDTGLGGGYEPKLEPTVILSRRRREATFTADGAIIYSIIAVNLHHNFITEEIVG IAPFLFINN (PAK 2077) SEQ ID NO: 5 MEHEYNEKEKQRNSAIKLNDAIRNNEENMDMTSPLELNFQNTNRKSRGLRERFSATLQRNLPGHSMLDRE LTTDGQKNQESRFSPGMIMDRLMHFGVRTRLGKVRNSASKYGGQVTFKFAQTKGTFLDQIMKHKDTSGGV CESISAHWISAHAKGDSIFNQLYVGGKKGKFHIDTLFSIKQLQMDGYLDDEQSTMTEYWLGTQGMQPNIQR NDDTDEHSSKVVGETGNRGTKDLLHAILDTGDKGSGYKKISFLGKMAGHTVAAYVDDQKGVTFFDPNFGE FSFPDKTSFSHWFTDDFWPKSWYSLEIGLGQEFEVFNYAPEAP (PAK 2892) SEQ ID NO: 6 MPNKKYSENTHQGKKPLMKSEANNEHDIQNSSLGIGLDLNSMMGNSSTSLSHIQDYSFWKENISEYYKWM VVVKAHLKQLDWTLKSMDSPESAGTNIAKNTGTTALQTLLNTGGSIAGAAIGGAIGSAIAPGVGTIAGMGIGA LAGTGLNYLNDTVIEKLNEKLEIAYPYPKTRNMIFDINNYDKNPIIKAIKKKTNKDNLKVTAGSSLTSQLVGKVT SPIKFPAYKLADLAIALAGLSSDKARHILDFTDSIREVLNESHSDAVAFMRKNYGDNAMGLAGLSSRIK (PAK 2893) SEQ ID NO: 7 MEREYSEKEKHKKRPIQLRNSIEQHEEETANNSLGLGLDLNQATNPPKVPKDNYNEENGDLFYGLANQRG RYIKSVNPNFDPDKINSSPMIIDVYNNNVSNTILNKYPLDKLVKLSGNPQKYANNIKVENSLQQDVASSKRGW YPLWNDYFKTGNENKKFNIADIYKETRNQYGSDYYHTWHTPTGAAPKLLWKRGSKLGIEMAASNEKTKIHF VLDGLNIQEVVNKQKGSTPLEQGRGESITASELRYAYRNRERLAGKIHFYENDQETVAPWEKSPELWQNYI PKNKNQNESSTPQRNNGTLYRLGGPFRKLRASLRKRS (PAK 2894) SEQ ID NO: 8 MMEHEYSKEEEKKRQQSKPNNATHDESNLPLELEKHFNARTPATAHSKWFTYENDTEVELTTERIKEIFSN KQPKIIIAGDGHNKPPFQYAKNIPDVNSSFDAGTLQLYIEATDEQINENNPEYIPKEFMAKPGLFTNKNRRAEI VGWEDSELSNAMKEMFELSDKSTREKLTPEETSSFYKLHETAIRHFFRPEFNQLRDEFFEILAKAGSNRELD KIALEMIGFTSGTWRDEYINPTLAEKIAKHAAEKENHTFVVSIGDAHLSENPMQEYLNKRRNGGEFKHQIIFT RDKRPILPDNMKTGNKNS (PAK 3525) SEQ ID NO: 9 MLKYANPQAVPTQRTKNTAKKPSSSSSFDGQLELSNGEWSKHSEMGLKRGGLINSIRRRIARNGNIGRFNE LIDSEAKKWPSEPVDKNIHMIWIGTRNISEKNIKLSIDTAKKNPDYNTSIIYDSGISGHEGARNFMLEKFEGSN VNXSLAFPKGIGVMREYAPEAGKATAFPNTPIAVTKNNPIINKTLDLAVGNYQRGEKNVLKLAGPDVFTQALY QEIPGLNSKVLNAQLDQFELAKRQALGLPLEKPKSFADEKLTSVEKEKINRPYQSMRGLSGHVMNGADHS WAVDTEVLGH (PAT 00148) SEQ ID NO: 10 MMREYSNEDDCTKEKTNLVKSENVEADNYLEMEHLTYLAKLISMTERENHHLNSIKLIDDIIELHNDRKGNKL LWNDNWQDKIIDRDLQSIFKKIDEMVSEFGGLEAYKDIVGESPYDPTEPVCGYSAQNIFKLMTEGEYAVDPV KMAKTGKINGNQFAEKLEHLNSSNNYVALINDHRLGHMFLVDIPSTNRERVGYIYQSDLGDGALPALKIADW LKSRGKESINVNKLKKFLNDEFTMLPENEQKGLIAEIFDLNKDIDSVKSGKIKKDKAVDIYLREYDINDFISNVE KLKTKLA (PAT 00149) SEQ ID NO: 11 MIFKMLNLAVFYLLGNIFHYLICQKFICYFCSVLKSVTMFLTKVAVQIALYLNILPTMAGIAGLHAEVQALNNLFI SGDRGTEKRENWKYIRNMLESTIFTQRLTAGQAGKDFAACHNCSGILSSPVNVITGKVESAGGNFFINIISI (PAT 00150) SEQ ID NO: 12 MEREYSEKPKNLSQLSRKTAISERRAMFERNASSNNEQPVPQFARSYTSNRSVVNINPGRSSIAVVTANST SPVNISTPAAASPDKLLPSTSCDTTSSTLTVGKYKLELTSQGKVVVFRGDNRTPEQIVAAGGFGEQSNYLYK MEIPGLKPQDMNERTLGEKIRQDSRGN (PAT 00152) SEQ ID NO: 13 MKYDPRLRTVWEDDFDYEKNFKKQTDYINYKDLEKQLKENVDYYALLDENEAIIFLKELGCDIKSFLNDTAFP VTDVLSNFAGNIKDALGVFKVAKNFKPINIGIFTYIINELKGKGIKAIEYLGKNGERYIKLTDRPGIRKYLNATRY LINNKKIMEVGIGSVAMEGSIVKGARFGVIYSAAYRSVELMFKSEYDLTNFFVNLSMDMAKIIVATIIAKSTVAA ATSFVVTAALSTTAIAIGVFIIGALVVWGLMWLDDEFKISETIIRRLKEHKVKTPISTYHSDQIFNAWGRYYRG (PAT 02308) SEQ ID NO: 14 MPNKKHSENTHQGRKPLIKSEANNEHDIENSSLGIGLDLNSTIGNNSASLSQIQDYSFWKENISEYYKWMVV VKAHLKQLDWTLKSMDSSESAGTNIAKNIGTTALQTLLNTGGSIAGGAIGGAIGSAIAPGVGTIAGMGIGALA GTGLNYLNDTVIEKLNEKLEIAYPYPKTRNMIFDINNYDKNPIIKAIKKKTNKDNLKVTAGSSLTSQLVGKVTSP IKFPAYKLSDLAISHNRALAGLSSDKARHILDFTDSIREVLNESHSDAVAFMRKNYGDNAMGLSGLSSRIKGE KLTLATLARTRNKIENRINSINKQTLKLSSKNSNE (PAT 02309) SEQ ID NO: 15 MEREYSEKEKHKKRPIQLRNSIEQHEEETANNSLGLGLDLNQATNPPKVPKDNYNEENGDLFYGLATQRGR YIKSVNPNFDPDKINSSPMIIDVYNNNVSNTILNKYPLDKLVKLSGNPQKYANNIKVENNLQQDVASSKRGWY PLWNDYFKIGNENKKFNIADIYKETRNQYGSDYYHTWHTPTGAAPKLLWKRGSKLGIEMAASNEKTKIHFVL DGLNIQEVVNKQKGSTPLEQGRGESITASELRYAYRNRERLAGKIHFYENDQETVAPWEKSPELWQNYIPK NKNQNESSTPQRNNGALYRLGGPFRKLRASLRKRS (PAT 02310) SEQ ID NO: 16 MMEHEYSKEEEKKRQQSKPNNATHDESNLPLELEKHSNARTSATAYSKWFTYENDMEVELTTERVREIFS NKQPKIIIAGDGHNKPPFQYTKNIPDVNSSFDAGTLQLYIEATDEQINENNPEYIPKEFMAKPGLFTNKNRRA EIVGWEDSELSNAMKEMFELSDKSTREKLTPEETSSFYKLHETAIRHFFRPEFNQLRDEFFEILAKAGSNRE LDKIALEMIGFTSGTWRDEYINPTLAEKIAKHAAEKENHTFVVSIGDAHLSENPMQEYLNKRRNGGEFKHQII FTRDKRPILPDNMKTGKKNS (PAT 02956) SEQ ID NO: 17 MSNYEYDIVTQHDTYQIKDNEYTVVNGKYWQYEQEGNKNNNKISISLMKDNQNDPVWITSDIKEISLYIIENL FSYHKFSAELQHTLKNAVKAVFNEYSEIKYSELLHNINNIFNLFFIKTYNTSDINTAINILTAKIEIYDKLEKINQD KTDLNNTKVDIWEELGINAEEPLLKIYRQAFSTGDIDDEVYSDALLTFMSDGNLKLGDKEKSDYNQRIKDKTD LFESYKKGIEKVASLITTNNINPGIPITYPETEKSINIGDDLLLAQLAKEEIALKKQNRTEYSQQDIFELQTLQAA KYHLLILSSLGALLYQIAPNVEKMTKGHGDYRDIIFSQEQAESLFKKHNIQYDTNHVLSQESKHIEMEGCIILTA AIlYRMRKENATVEQALNYSTLETIKLFENDKKKLNPFNTNNVKPAGYFSFIDFKKRDKFDSQYNFNEQFNVY KNKYSHYESISFSKLILSSPAAQLTAEEIVNPPEETFLYSVEQGMGNVAMIKMYQGNWLVVSTIQGGVKARK YSQQQVDSQPTLRAMSRPNALFLIERKIMIGIGIFMENQIVNTGKRLFPTGYERAKTLSGFAETSRYKNSYNA FWNDYYGITSGMNVGISFTGSPKFNFYKEENLLSVTATIIQQGLNDIAIKSKQALDITSGWHIAATILIPFYNVIY KSTTDSEYELTGEDIGSIVFDTANVLLVVATLGMSLTESMAAKVTQTTLRLRQAGLTGRALITAVVRTLPEHGI ITLRQSSGIILGGLIDLIEPLPIRSTLTLTYRGVISAVGAMRNSIKLEKSFADIFGKSTRGLGKLKHEWKVSNLPL EEIVPHSNGGEIYKGIYSIRHTNPETAVKQNFYIKEAGANYQVKWDDANHTWRVVNPTYPEQFSYWPAVKL DKNGHWWTHADISNKFLILEKSKRIDQELEAAHSNINNDNILDAFIHINTAFKDCERYDIDKLSDITDTLTHFFE KSLKPGDKKAIFSTEIMSIQQAWIREVILPLQNNSSISIEKINAIKTELPYLLRKTFPIESQLPNQLVANKIALAIEE IPNTRIPKYTSGNISKTVQYTSLLENNHVDIPPVGITITGNDTFINQVTRVLSEIDEIPSGNIVIQELEKQGLNIQP PTMNDIVREKNGQFYANNSAGSHIAFDPENHLIGTEEKLIDEPWRTREPAIALYHEMLHIYYNRYPTWFTSID NKVIDQKVSGGFSLLEESRIVGTKYYVNDKDTLFDFNDSDYLLENNSALLTENRFRAEYAIFKNKSEYVIRPY SGKGDSQIPLTKTKININESHRNVMGVGSGKPEKMPNESATDYRNRVREWRKANKQPEADIGTGDMRKTK AEARVKLLKENYPQFEPQKIELGGAFQLWTVPNEPANKLMLSSHGYFFSDSAATQVPAGKTIQFLGPHGKT LLEAPENPLNSPFDVTLGNSGFTVQPYATIESGNKAGLGSVKIGDKTFTVNDIQNIATDDVENYLLATGVEAN ASNHGKVRNYGIKYYEKMPDEEVKAAIWKNRADETSTHKYDALLVSPEAGNRKKLSDIFALMKTDERMSKY DEITFVACREELNRINMKSIHDTGLGGGYEPKLEPTVILSRRRREATFTADGAIIYSIIAVNLHHNFITEEIVGIA PFLFIDN (PAT 02957) SEQ ID NO: 18 MEHEYNEKEKQRNSAIKLNDAIRNNEENMDMTSPLELNSQNTNRKSRGLRERFSATLQRNLPGHSMLDRE LTTDGQKNQESRFSPGMIMDRLMHFGVRTRLGKVRNSASKYGGQVTFKFAQTKGTFLDQIMKHKDTSGGV CESISAHWISAHAKGDSIFNQLYVGGQKGKFHIDTLFSIKQLQMDGYLDDEQSTMTEYWLGTQGMQPNIQR NDDTDEHSSKVVGETGTKGTKDLLHAILDTGDKGSGYKKISFLGKMAGHTVAAYVDDQKGVTFFDPNFGEF SFPDKTSFSHWFTDDFWPKSWYSLEIGLGQEFEVFNYAPKEP (PAT 03171) SEQ ID NO: 19 MFKYDTSEKMAKFGKGKTSDGMLLDTLYLEIPDEKAVMSAYKSQILDELRNFSEKTHSFFSGKKPLYSKKYL ANLAAHAGYVHVTDYNSIGNYKDGFVNFKDNSRNLAEGKLFPGIRLIKRPKLSIVRDKETERWKKQESDEAD AYEITDIESFISGVRDMYSRANVDLHPVIESLIRNHIVNNDHVLPTMAGIAGLHAEVQALNNLLILADGRAGKIV GGRKIEEYMQDMLKSFIFTQRLTTKQAGNDFAACHNCSGILSVPANVITGKVASAGSNFSLILSRYKNSQES PI (PAT 03172) SEQ ID NO: 20 MLKHANPQTVSTQRTKSTAKKPSSSSSFDRQFELSNSENQPGEGNKDWTIKGWRQRFADRSLNKGHISPL MNKGLLVGSEEALINVPVVAHRYDSSHQLTDAGPLKADSHSNNLDPFYGVVTGFRGDQVTSSESGSGSIG GHWGKNTLDSNITGINVVNGASGTVGIRIALKDIQHGAPVIVTSGALSGCTMVYAVKNGYFFAYHTGQKPGD KEWKTGRQGVVATYRSHQALSPDSEPMAVGEQNNDLVNIFASYDQGIITYMGKPGVIIDNTAENVGVFNYD EVKLEKPDIRAGYSYALLAKDDKGKVNVKVLSEDVIVPLGNKGKTIKAINSLKKRLL (PAT 03177) SEQ ID NO: 21 MPRYANYQINPKQNTKNSHGKSSSSNFSSGYFSSSNNSLDDSLIRQQVKREFIWEGHMKEIEEASRLGNFA VSFRAAGGPTLRALGKGAAAKGHDILEKTIKPGSINKAYPKDEASNVIKKVQEAGIEGYVGHWDKKTGRLLGI YMSSGHGLSDEQVNGKIYPIDLNNLEASLSALKTKENWAALPFTGDYDMHDMISFTGQPHSVPSNSSEERK HDRINRLVARSDPNRPFGDIEHNVIRHGAQVSYPAFAMDKEKEEIKKHGGIVKAVAEPGEFPVAIVSKGKWTI ANNIDELNQFYNSIGAKMKVSWKPGAENPGFVSNPQRPGMARFSRKR (PAU 02009) SEQ ID NO: 22 MMREYSKEDDCVKEKTNLAESENVEADNYLEMDCLNYLAKLNGMPERKDHSLNSTKLIDDIIKLHNDRKGN KLLWNDNWQDKIIDRDLESIFKKIDEMVSEFGGIEIYKDIVGENPYDPTEPVCGYSAQNIFKLMTEGEHAVDP VKMAQTGKINGNEFAEKLEQLNSSNNYVALINDHRLGHMFLVDIPSTNREKVGYIYQSDLGDGALPALKIAD WLKSRGKESINVNKLKKFLSNEFTMLSESEQKELIAEIFDINKDIANVKLGKIKKDKAVDVYLREYDLNDFISNI EKLKTKLV (PAU 02010) SEQ ID NO: 23 MPIIGHKEDLIRTERSSVDLTRSSNNRQTDNLELNIPQHKRDNKDIEHAVIYGFSQHRGPEMQKAFADNKNP VTIDEYNAGLGIMGELSLSDYFRISQDLKENRLPELNEKNIQNHSLKYFDAMGVNMKSADPNVKEEAKEQQ RAYTRSWGFYMMENKEKLDIQSKINNLIPKKKSFFSKSPGEDEYKKLDEFILKNSNGSNLTIPKQRKILMKFA SAKNAVDVTKNLSGEEQTWLKDIIATAFFRQTSKLGMSWFIEQLASPDFRFVIVGFNGEELTTDQIRSNKPW KHGNRRKEGASEYAEPITFSEIRHAHRKGYDSKINFIKK (PAU 02095) SEQ ID NO: 24 MISTFDPAICAGTPTVTVLDNRNLTVREIVFHRAKAGGDTDTLITRHQYDLRGNLTQSLDPRLYDLMQKDNT VQPNFYWQHDLLGRVLHTVSIDAGGTVTLSDIEDRPALNVNAMGVVKTWQYEANSLPGRLLSVSEQSANE AVPRVIEHFIWAGNSQAEKDLNLAGQYMRHYDTAGLDQLNSLSLTGAHLSQSLQLLKDDQMPDWAGDNES VWQNKLKNEVHTTQSTTDATGAPLTQTDAKENMQRLAYNVTGQLKSSWLTLNGQLEQIIVKSLAYSESGQK IREEHGNGVVTKYSYEPDTQRLINITTQRSKGHVFSEKLLQDLLYEYDPVGNIVSILNRAEATHFWRNQKVSP RNTYTYDSLYQLIQSTGREMADIGQQNNKMPTPLVPLSSDDKVYTTYTRTYSYDRGNNLTKIQHRAPASHNI YTTEITVSNRSNRAVLSHNGLTPREVDAQFDASGHQISLPTGQNLSWNQRGELQQATTINRDNSATDREW YRYNAGSARILKVSEQQTGNSTQQQQVTYLPGLELRTTKSGTNTTEDLQVITMVETERTQVRILHWSAGKP NDIANNQVRYSYDNLIESNVMELDTKGKIISQEEYYPYGGTAIWTARNQIEASYKTVRYSGKERDKTGLYYY RHRYYQPWLGRWLSADPAGTVDGLNLYRMVKNNPIRYQDESGTNANDKAQAIFKEGKKIAINQLKIASNFL KDSKNSENALEIYRIFFGGHQDIEQLPQWKKRIDSVIYGLDKLKTTKHVHYQQDKSGSSSTVADLNVDEYKK WSEGNKSIYVNVYADALKRVYEDPLLGREHVAHIAIHELSHGVLRTQDHKYIGVLSSPGSHDLTDLLSILMPP ANEQDRTEKQRRATGARKALENADSFTLSARYLYYTAQDPNFLSSLRKAHRDFNNKKTDRLIIRPPERR (PAU 02096) SEQ ID NO: 25 MEREYNKKEKQKKSAIKLDDAVGNNEENMDMTSPLELNSQYTNRKRPGLRERFSATLQRNLPGHSMLDRE LTTDGQKNQESRFSPGMIMDRIMHLGVRTRLGKVRNSASKYGGQVTFKFAQTKGTFLDQIMKHKDTSGGV CESISAHWISAHAKGESIFDQLYVGGQKGKFHIDTLFSIKQLQMDGYLDDEQSTMTEYWLGTQGIQPNRQK NDNMNEHSSKIVGETGTRGTKDLLRAILDTGDKGSGYKKISFLGKMAGHTVAAYVDDQKGVTFFDPNFGEF NFPDKVSFSHWFTDDFWPKSWYSLEIGLGQEFEVFNYEPKEP (PAU 02097) SEQ ID NO: 26 MVYEYAKTNDRKRKLSTQSDNYEEKSFSPVLDLSRNNQNTPNMEDEYETPQNFINRTGREKLFRAIRMVAS NKRDPITKDQVSVPPDGNLFTELKDKHLDRAAEYKKLKTWPTHASIIATSPSANTPIAQHVSGDDALSPYIST GDKPGAVQNTVRNWNGIGPASERRLRPEKTWSPIIEIDVNKLPDTTKIFDLNKPNNTFFSTTNSDIAQNAFAD KEVLISPEIPGLAITRVINDPEEIKQIANLNPSQSLIEKKNTIPEEKIIFEEKKSVPIHDSDADIPSSSFVFPKRKKP RNIRSRTDS (PAU 02098) SEQ ID NO: 27 MVFEHDKTVERKRKPSIQLGNDKEKSSEQALELPQSKQNNPLLHDLITSNNLRKEAAVFAKQIGPSYQGILD GLEHLHNLSGNEQLTAGFELHRRITRYLEEHPDSKRNAALRRTQTQLGDLMFTGTLQEVRHPLLEMAETRP AMASQIYQIARDEAKGNTPGLTDLMVRVWKEDPYLAAKSGYQGKIPNDLPFEPKFHVELGDQFGEFKTWLD TAQNQGLLTHTRLDEQNKQVHLGYSYNELLDMTGGVESVKMAVYFLKEAAKQAEPGSAKSQEAILLNRFA NPAYLTQLEQGRLAQMEAIYHSSHNTDVAAWDQQFSPDALTQFNHQLDNSVDLNSQLSFLLKDRQGLLIGE SHGSDLNGLRFVEEQMDALKAHGVTVIGLEHLRSDLAQPLIDKFLTSENEPMPAELAAMLKTKHLSVNLFEQ ARSKQMKIIALDNNSTTRPAEGEHSLMYRAGAANNVAVERLQQLPAEEKFVAIYGNAHLQSHEGIDHFLPGI THRLGLPALKVDENNRFTAQADNINQRKCYDDVVEVSRIQLTS (PAU 02230) SEQ ID NO: 28 MKGIEGVIMLSHDILPEKLLVSEKKHENVGSYFSDDIGEQSEQTEVSHFNLSLDDAFDIYADISIENQQELKNK DNNTNIWSSLGRGDDDHNLKKIINDAFKEKLPQLMEYRRKGYNVIGLDKEGIKKLEGMLKAVPPEIQQPTMK NLYSAAQELLNTLKQHPLLPENQDMIQQSNLVIRNLSDALEAINAVSKVNQVEVWVEEVHKTNKAQSDRLIAA TLEELFFKVKDKRLPGSNDDYCQQEREETERKIKDLLLYDGYQLTAEHFKFGRLRKSLLAESRVTRLKLAEY LEKKSVGILTAARDAKMYAMKILLAQTRNNGFNAKDLINAGQVNDRLLSFQQYARHIRAVDGEIDGIILSNPLV VACIKETNDEPAHIKIARAILPVSEELGTVSKVLRETKEKVQPSKPKEELNHPHQDVWVNRGDELWKYIKKTS WNIKETSVHVTQMVGYEASKTASRAKHKLKESSYSESINGAVKGTALLLLDEIQQAENRIRQIPQFAWDVQE AVEQHSSVIQRTAYPDELPELSELLNEQLKHEEARWQAVKKQSRDKLQELIAPITRLAQEKWAQDLYFQLG EELRKERQDRWKDIQQFDEIMAEAVGQFAEMARELDSEAVRLAEHGHSGGKELQEKVAKWLRDLSKLKGK VKAGVAKITGTSLDNFSRSGMLARGMSEWAEDLKQSYLQETLQEGSAVAAELFERTLMEVVEENRTHFAK ESDPEAERFLKRLALALKHAAENTTVYPPTPEEILAGSRSLPEDIRHWAEKKVVSGAISAAFRGGFKLVTGTF SLPVRVVIRGAKTGGTLYRGVRAINRSVRLGQGPATQVKSKFINQELSKTAFRLTLSLSPLVAWGMAASITA GRLYNEKDYPEKIIKNIVIDLPEELLWIGGYAGINAAIRAHAEKAIQQAIQHALDEQADKLALRINKEIAGKSADV NVEIIPQETSVSPAETAQSTPEPLSDFASTSQLTMPELIDIQDNNSAQQPKVRRKRDVSVESEISIDNLNIINA NTREDKVNSEIKSELRSELKRFENSDANSPMSDVERAIFIDLFLYKNKYEVSESQQDYKNTWLKFRRELESQ ENKEIKEYLRFRSIIEAYEIYDKKRLDDDTIPEAGTIIKEVIDFFQKLKKENPITFMKLAEAMVKFQYYYEEEDEN EDRYFKMAEIYYFLNKTENEKKSKTFHLDIIDKYPNENNRLLDEFFLNKNNNNPDLDEIIYKLQSMQEKYRES YEMLSKVENIHQVLSDDSKNEENIFLDNRIIAAQVFDGSINISLQDKKKWLNRYDQIRNEEGSDGWKLMHIES ILINLRRINTAINLTAMKSESALLLIDKLLNFQKKARENILHISETPHEDFTSYSQFKTRKELGNDDSKYYAQFD NYKDNHDAEKEAKEILSQVVARASLSFSELFDKVESIKLFSFVYKNRDGGAPLAAPGRTVVIKFPGKDTGGL VISNLFLRNHVKRISTKEMEDLKPLTEGMYTRATQHRSLGSYYHIGSQSEHTNALEILSGMNKEELKTHLKK QGIWFGEPALFSNEYPKQENTGHLENTTLKNAIIGVSTIQNNAAANYLRSTMYESTGWEKLGDRFIPFYEIGR RKHYDREYEINSEQLTLDIITSIAIAYPAARGIVATIRSSAIPSILKSGLRGSALFKSLSLELGKMGFNASKVFGG AVYELIEPYPINSHLNRHNVFNKVKDTAWEFHTDVGLKGGGLKDFIDRFTKEPKEITISGYKFKRIKYNQENF DTMQRMALDYAYNPDSKGKIAQAQQAYKTGKEDYNAPQYDNFNGLSLDKKIERYISPDTDATTKGVLAGK MNESIKDINAFQTAKDAQSWKKSANKANKVVLTPQNLYLKGKPSECLPESVLMGWALQSSQDAKLSKMLM GIYSSNDITSNPLYKSLKELHANGNASKFNASATSISNINVSNLATSETKLFPTEISSVRVDAPKHTMLISKIKN RENKIKYVFYDPNYGMAYFDKHSDMAAFFQKKMQQYDFPDDSVSFHPLDYSNVSDIKISGRNLNEIIDGEIP LLYKQEGVQLEGITPRDGIYRVPPKNTLGVQETKHYIIVNNDIYQVEWDQTNNTWRVFDPSNTNRSRPTVPV KQDTNGEWFKHSETGLKGGGPIDDIRKYIARKSAIKIFNQSINYSATKWPPEPIDKNIHMIWIGTKNISEKNIKL SIDTAKKNPDYNTSIIYDSGISGHEGAKKFMLEKFQDSNVNIIDFRKKSYFSQLKQEPSFAYYEQVIAENKYAQ ASDILRLLVLKYEGGIYKDIDDIQVKGFGSLTFPKGIGVMREYAPEAGKATAFPNTPIAVTKNNPIINKTLDLAV SNYQRGEKNVLKLAGPDVFTQALYQEIPGLDSKVLNAQLYQLELAKRQALGVPLEKPKNFADEQLTSAEKE KINRPYQSIRGLSGYVENGADHSWAVDTNIPSTSTQTSTIVTPLAPKTEMLPPVPSSSTKSSTSAPVLQEKIS YNLATDIDATDYLNQLKQKTNINNKISSPAGQCESLMKPVSDFMRENGFTDIRYRGMFIWNNATEQIPMNHF VVVGKKVGKDYVFDVSAHQFENKGMPDLNGPLILAAEDWAKKYRGATTRKLIYYSDFKNASTATNTYNALP RELVLESMEGKTFITSPNWYQTFKRTHNIHPEVTVSDPATFSLNYSVNPTAENLSPPPPPPIPSHGQVPKTV TPPPPPMRSPLSLSQPLERLPANKTKPIGFNPGENKASFSKLEEAGKHYYKDDKSRQAAPVNTMSDFDNRY LSHTTEAPAPSNVAHLAPGNIYNTKVTAKGAEKPAYDIYISKDGESLITSSSYKVDDITTDSKFGKPLPYSEIM FNSLKKSGVDPKNLKRSVQASIENKVTQDVISAIGTRIQRGQVIRVSPTENPDAFYTLLGTDNCKATLHMLNQ HAEEFGHKWTSIEFKGTGYLVMNIGTSTQTSTIVTPPPMPGTSQLVQ (PAU 02805) SEQ ID NO: 29 MPNKKYSENTHQGKKPLIKSEANNEHAIDNSPLGIGLDLNSILGNNSASLSQIHDYSFWKENISEYYKWMW VKAHLKQLDWTLKSMDSPESAGANIAKNIGTTTLQTLLNTGGSIAGGAIGGAIGSAIAPGVGTIAGMGIGALA GTGLNYLNDTAIEKLNEKLEIAYPYPKTRNMIFDINNYDKNPLIKAIKKKTKKDNLKVMAGSSLTSQLLGRITPI KIPAYKLADLAVSHHRALAGLSSDKARHILDFTNSIREVLNESHSDAVAFMRKNYGDNAMGLSGLSSKIKGD KLTLDTLARTRNKIENRINSINKQTLKLSSKNSNE (PAU 02806) SEQ ID NO: 30 MEREYSEKEKHKKHPIQLRDAIEQHAEETANNSLGLGLDLHQAINTPKVPKDNYNEENGDLFYGLAAQRGR YIKSVNPNFDPDKTNSSPMVIDVYNNHVSNTILNKYPLDKLGKLYGNPQKYAKDIKVTNSLQQDVAASKRGW YPLWNDYFKAGNENKKFNIADIYKETRNQYGSDYYHTWHEPTGAAPKLLWKRGSKLGIAMAASNEKTKIHF VLDGLNIQEVVNKQKGSTPLEQGRGESITASELRYAYRNRERLAGKIHFYENDQETIAPWEKSPELWQNYIP KNKSQNESSTPQRNNGALYRLGGPFRKLRASLRKRS (PAU 02807) SEQ ID NO: 31 MVHEYSINDRQKRHSFSSANPIDPEVTNRENSRHRFPKDNYNKGHGDLFYGLAPERGKYIKEANPKFDPNN PENAAMIIDVYNDEISRVILNNNANKISTNRLLNFIYNFRKNRLENLMKNPEKYAKDIKVKDNLRENISPKKIEK YPLWNDYFEAGIRNKKFNIAEIFKETASQYNSDYYHAWHIGGNSAPRLLWKRGSKLGIEIAASNQRTKIHFIL DGLKIEDWNKTKGPAPLKAGPGESITASELRYAYRNRARLAGRIHFYENGKETIAPWDKDPELWQKYTPKN RSGMEL (PAU 03332) SEQ ID NO: 32 MLKYANPQTVATQRTKNTAKKPPSSTSFDGHLELSNGENQPYEGHKIRKIKGLRQHLADRSLNKGHISPLM NKGLLVGSKDVSIDIPVIAHRYDSSHQLTDAEPLKADSHSNHLDPFYGVIAGFRGDQVTSSESGSGSIGVHW GKNTLDSNIMGVNVVNGASGTVGIRIALKDIQHGSPVIVTSGALSGCTMVYSVKNGYFFAYHTGQKPGNNE WKTGRQGVVATYLSHQALSPDSEPMTVGEQNNDLVNIFANYDQSVITYMGKPGVLIDKMAENVGVFNYDEI KPEKPAIRAGYSYALLAKDDKGKVNVKVLSEDVIVSSGKQGNTVKAINSLKKRLL (PAU 03337) SEQ ID NO: 33 MPRYANYQINPKQNIKNSHGKSSSSDFSSGYLSFSNNSLDDPFIRQQVKREFIWEGHMKEIEEASRLGNFA VSFRAAGGPTLRALGKGAAAKGHDILEKTIKPGSINKAYPKDEASDVIKKVQEAGIEGYVGHWDKKTGRLLGI YMSSGHGLSDEQVNGKIYPIDLNNLEASLSALKAKENWAALPFTGDYDMHDMISFTGQPHSVPSNSSEERK HDRINRLVARSDSNRPFGDIEHNVIRHGAQVSYPAFAMDKEKEEIKKHGGIVKAVAEPGEFPVAIVSKGKWTI ANNIDELNQFYNSIGAKMKVSWKPGAENPGFVSNPQRPGMARFSRKR (Plu1651) SEQ ID NO: 34 MPNKKYSENTHQGKNPLMKSGANNEHDLQDSPLGIGLDLNSMLVNSSTSLSQIQDYSFWKENISEYYKWM VVVESHLKQLDWTLKSMDSPESAGTNVAKNMGVTALQSLLNTGSSIAGGAIGGAIGSAIAPGVGTIAGAGIG ALAGTGLNYLNDTAMSKLSKKLEIAHPYPKTRNMILDINNYDKNPIIKAIKKNVNKDNLKVTAGSSLTSKLVGT VTSPIKFPAYKFAELAVSHHRALEGLSDDKARHILDFTNSIREVLKESHSDAVAFMRKNYGDNAMGLSGFSS KIKREKLTLNTLAKTKNEIENRINSINKQTLKVSSRSRNE (Plu1671) SEQ ID NO: 35 MLSTEKHNKDTKHPRNREKKFSIQPENSTQDDEDIKNNSLGVGLDLDQMIRNTSSTLTNAPQKPEDGYYYHI SRGNNLQSFLQNGFKPQGSPGPTLSEEDFSRRKIGIIKLIYSIIATTINKNRKAKKISKDNFLMPQEFWHEFKN FYQNIPTQTNIDDQLLKKSITESIDKLDQNKFMEKHSDRKQTIINNEREAILQQDERINEIISSRAKMIQQREAE NTEGYIYLAPHKNTLLEYMKHLQEEKNLFLILAVKEDIFTEKGLEQDPQEPHGAVRYKGALSTEELNFVNQE GQICAIPASIGEMDYGDFILNQQQVIDFCKK (Plu1672) SEQ ID NO: 36 MPINDLKKKFEISPQAAQAIGAPARSNSSKQAEHQTEHLELDTSKNRRDRKDLNAQATPNQQHTKKLETEV NNGGNKSKAQAHTPDLVMKKESSVTPNTRKSPNEKIKAEDIFHRYKDRFSPSDRELPFEIMNEITNNGIAFS SEKAPESHLDKVKDKKFTLRHYTSGNGQEKPTFNEIGSNFNLVNEGIKTLKRTQGSNTNEDDWNRLGNTAF TFFLLAIDGEVSDRKFLSNTTHFAEIDIENPAELKELGLDETEFFASPDLLHEKNLSQAPAVKGKLSDLKSLLL KQSGIKPVQLQSLGAKGILERIDSKFNGSLEIKIPGNVKVKEWKKVEK (Plu1690) SEQ ID NO: 37 MPNSKYSEKVNHSANGAEKCSIHSNQYNINNCTLGLGLDLNKKLRTGNERNIEGAQPFIPFPSKQKQYSTSP IAMADILNESALTSQPIITDLINPQKIKMSDGVKNILNNKEGGGDLVFKALQIKPSDETLPFNALKIVDTYQEEM PNKDMSISAYWAPQGGYVDIPAQPDISRHPQYVFTPNFSGCSFVVDKMNEDTLRVRHVQGGQEDVEYNN QNIDHGMGMITAMEFRDYGYHEADDKVIENTYGFAFLKFNQEKKQWQLHYQKIAAAPNIINIKTKSSWLPFS KPSIEADTFTFKNMKVPGYSRKNINNN (Plu1691) SEQ ID NO: 38 MPKLTELLSRFENPIQNQPNHISKKNPISNSKVLNNSEEKTAPLELKHDDSKIKSQVSIPNLVKKNEKPAASNT PNNSHEKVKAEDIFNRFKSKFDPYDRELPFDIMNKITNNEIKFSSEKSKDDYLAKVKDKKFTLRHYTAGTGQE KPTFDEISSNFNLVNKGIKTLNRTQGSNTNEDDWNRLGNTAFTFYLLAIDGEVSNRKFLSNTTHFAEINIEDS EELKELGLDQAEFFASPDLLHEKNLSQAPAVKGKLSDLKSLLLKRSGISSVQLGRLDAKAILKSIDNEFGNSL EIKIPGNVKVNKWNKI (Plu1712) SEQ ID NO: 39 MPRYSNSQRTPTQSTKNTRRTSPSSNSSTEHLSLSNAPTNDSSVRQEVKEKFIWEGHWEGHMEAIEKASIL GNFAVSFRAAGKPTLEALGKGAAAKGHDILEKTIKPGSIEKAYPENEASDVIKKVREAGIEGYVGHWNKETG RLEGIYMSSGHGLPNGQVNGKIYPIDLNNLEASLAPLKEKKNWAALPFTGDYDMHDMISFTTQPHSVPSNS SEEKKIIDRINEYIAKSDSNRPFEDIEHNVIRHGPQVSYPAFAMDKEKKEIKERGGIVKAVAEPGEFPVAIVSK GKWTIANNINELEQFYNSIGAKMKASWKPGAGNPGFVSNPQKPGMARFSRKK (Plu1713) SEQ ID NO: 40 MFSTYSSKNDNQTINKINTEEKHENTETDNHLEINLEHTGKSKPDIEPKDVTTGTINAGTLLYKTTAIPEFLDN AKSLGLAEYEKRHKDIQDYLNLGKAEDAEKLKNKSQWAGQYFALEKSYDEYANEAPDSYNNLLKNAGKDLL ENTEEVKVFLYTFKVTKDIKVLKPHNNSNSYYVGDTEGWEKAKEIMNDVQSQSEKNDNPFPELKNLEDKNF LLEELGEKGYAWMGPLHAKEGAEKGTEFSYELAISPNLLRQHLTLESEELLGTYKNRYGYWDKK (Plu1714) SEQ ID NO: 41 MKKTDEKYGQYEYKDEDITSYPIAWTNPDNGKIYIGINSPEYSHLNNKGESELNLAKIISTIIHESLHASSHQH KGLQSQTDTGADNLNYDEYVTDYFAREVYKQILPDKDYVANCFTKGLGGENKIWGGNIVEFMIQ (Plu2400) SEQ ID NO: 42 MVYEYDKTIERRRNPSIQLNNNEKSSEQALELSQNNPLLHDLITSNNLRKEAAVFAKRIGPSYQEILDELEHL HHLSGNEQLAAGFELHRRITHYLEEHPDSKRNTALRRTQTQFGDLMFTGTLQKIRHSLLEMAETRPEMASHI YQIAREEVKGNTPGLTDLMVRVWKEDPYLAAKTGYQGKIPNDLPFEPKFHVELGAQFDDFKKWLDTAQSK ELLTHTRLDEQNKQVHLGYSYNELLDMTGVESVQMAVYFLKEAAKQAEPGSTKSQEDILLHRFANPTYLAQ LEHSRLAQIEAIYHSSHDTDVTAWDQQFASDALTQFNHQLNNTVDLNSQLSLLLKDRQGLLIGESHGSDLNG LRFVEEQMEVLKAHGVTVIGLEHLRSDLAQPLIDKFLASGNEPMPAELAALLKTKHLSANLFEQARSKQMKII ALDNNSTTRPTVEGTQHGLMYRAGAANNVAVERLRQLPAGEKFVAIYGNAHLQSHEGIDHFLPGITHRLGL PALKVDENNRFTAQVDNINQRKRYDDVVELPRIQLTS (Plu2401) SEQ ID NO: 43 MEHEYSEKEKPQKCPIQLRDSIEHDKEDINTTTPLELNSQYTNRKRAGLRERFSTTLQRNLPGHSMLDRELT TDGMKNQESRFSPAMIMDRMMHFGVRTRLGKVRNSASKHGGQVTFKFAQTKGTFLDQIMKHKDTSGGVC ESISAHWISAHAKGESIFDQLYVGGQKGKFHIDSLVSIKQLQMDSYLDDEQSTMTEYWLGTQGIQPIMQKND VDEHSSKVVGQTGNKGTTDLLRAILDTGDKGSGYKKISFLGKMAGHTVAAYVDDQKGVIFFDPNFGEFSFP SITSFSRWFTDDFWPKSWYNLEIGLGQQFEVFNYELKKS (Plu2514) SEQ ID NO: 44 MYDSKKKNSEPTTKKKFERSNYSQWDDSINHYEDMNRARIKNRNDILTTVDYFGEKKKTMHTFEYQSDIKH DTNFNNKNKSLFESFAASFVLQNPSFFSGVIDKLSKKLFNIISKIDERNNFQKKLYDFIEKDTSPEGQFGRFTL GKNEILNVLQVKSDTPQLFVKKMLLIKSLGAFIIDFSSKDIGNYDFIFDGKGREVNDIIEKNRPTNLFKVRGRTN IKSSQHRSDIGILDTPTFDSLTEEQKSFLTIPELTKRRPLFRTFTHELDAEDKRVVESVFVNRTFDCDSPLIGS VSGSTSCVLVAADILFPDMTMVERKKLAIATFAFLVGGGYHSATEVFDVAYPGLDLNKEIEELIENNPIQENA GVATLRQLIGNSGF (Plu2515) SEQ ID NO: 45 MPISNLAKESEVRAVKDIPCKNIETDNHLEIGLSSGLSRSKDTSKFKKNSINTIKLIDDIIALHNDPKGNKLLWN DNWQDKIINRDLANIFEKIDESVSELGGLEMYQEMVGVNPYDPTEPVCGLSAQNIFKLMTEGEHAVDPVEM AQTGKIDGNEFAESVDQLSSAKNYVALVNDRRLGHMFLIDIPSNDQETVGYIYQSDLGQGALPPLKIADWLN SRGKDAVSLNKLKKLLSREFNLLSDDEKRALISETLDIHKDVSNVELDRIKRDRGVDIYLTEYDVNNFYENIET LKSKLSNYDKKLSKPK (Plu1649) SEQ ID NO: 46 MLANVLPNLASFLKYEKETPLFFIEDGFNFQNLNPGRVPLIKTPEQRKAGDTQSPAFLCSGVILRGTIHSNDY KFWQPSPSSIKSGGVSFSYLRKDAKFKRLAYGYKNGFIIFPEHIAPEDRVDFSVLCAFPIDGYTNERANQGC GENITKAKDKGKSCQEQNVTNSDDWIKNYRKVNSQDFFQCGFNVTKDVNNPAIAFYQMLESIKKLPRTPNT PPKQNEIRISTWEESDPNKLPIEALFYSENSGLADAQKDQRDYKNATGKFLPIVKMLLPRTLNEDALFKFNIK DQVINP
TABLE-US-00005 Leader Sequences (e.q. with SEQ ID NO: 47 - 92 correspondinq to amino acids 1-50 of SEQ ID NO: 1 - SEQ ID NO: 46, respectively) Sequence ID Amino acid sequence SEQ ID MMREYSNEDDFIKEKTNLVKSENVE NO: 47 ADNYLETEYLTYLAKLIGMTERENH SEQ ID MFQNRIRNEKTTQSGKGKTLDRMTD NO: 48 SLYLEIPNVEAVTLAYQKLTSKYRK SEQ ID MEREYSEKQKNPSKLSRKTAISERIA NO: 49 ALERSGLSNSNQPVPQFARPYTSN SEQ ID MSNYEYDIVTQHDTYQIKDNEYTVVN NO: 50 GKYWQYEQEGNKNNNKVSISLMKE SEQ ID MEHEYNEKEKQRNSAIKLNDAIRNNE NO: 51 ENMDMTSPLELNFQNTNRKSRGLR SEQ ID MPNKKYSENTHQGKKPLMKSEANN NO: 52 EHDIQNSSLGIGLDLNSMMGNSSTSL SEQ ID MEREYSEKEKHKKRPIQLRNSIEQHE NO: 53 EETANNSLGLGLDLNQATNPPKVP SEQ ID MMEHEYSKEEEKKRQQSKPNNATH NO: 54 DESNLPLELEKHFNARTPATAHSKWF SEQ ID MLKYANPQAVPTQRTKNTAKKPSSS NO: 55 SSFDGQLELSNGEWSKHSEMGLKRG SEQ ID MMREYSNEDDCTKEKTNLVKSENVE NO: 56 ADNYLEMEHLTYLAKLISMTERENH SEQ ID MIFKMLNLAVFYLLGNIFHYLICQKFIC NO: 57 YFCSVLKSVTMFLTKVAVQIAL SEQ ID MEREYSEKPKNLSQLSRKTAISERRA NO: 58 MFERNASSNNEQPVPQFARSYTSN SEQ ID MKYDPRLRTVWEDDFDYEKNFKKQ NO: 59 TDYINYKDLEKQLKENVDYYALLDEN SEQ ID MPNKKHSENTHQGRKPLIKSEANNE NO: 60 HDIENSSLGIGLDLNSTIGNNSASL SEQ ID MEREYSEKEKHKKRPIQLRNSIEQHE NO: 61 EETANNSLGLGLDLNQATNPPKVP SEQ ID MMEHEYSKEEEKKRQQSKPNNATH NO: 62 DESNLPLELEKHSNARTSATAYSKWF SEQ ID MSNYEYDIVTQHDTYQIKDNEYTVVN NO: 63 GKYWQYEQEGNKNNNKISISLMKD SEQ ID MEHEYNEKEKQRNSAIKLNDAIRNNE NO: 64 ENMDMTSPLELNSQNTNRKSRGLR SEQ ID MFKYDTSEKMAKFGKGKTSDGMLLD NO: 65 TLYLEIPDEKAVMSAYKSQILDELR SEQ ID MLKHANPQTVSTQRTKSTAKKPSSS NO: 66 SSFDRQFELSNSENQPGEGNKDWTI SEQ ID MPRYANYQINPKQNTKNSHGKSSSS NO: 67 NFSSGYFSSSNNSLDDSLIRQQVKR SEQ ID MREYSKEDDCVKEKTNLAESENVEA NO: 68 DNYLEMDCLNYLAKLNGMPERKDHS SEQ ID MPIIGHKEDLIRTERSSVDLTRSSNN NO: 69 RQTDNLELNIPQHKRDNKDIEHAV SEQ ID MISTFDPAICAGTPTVTVLDNRNLTVREIVF NO: 70 HRAKAGGDTDTLITRHQYD SEQ ID MEREYNKKEKQKKSAIKLDDAVGNNEEN NO: 71 MDMTSPLELNSQYTNRKRPGLR SEQ ID MVYEYAKTNDRKRKLSTQSDNYEEKSFS NO: 72 PVLDLSRNNQNTPNMEDEYETP SEQ ID MVFEHDKTVERKRKPSIQLGNDKEKSSEQ NO: 73 ALELPQSKQNNPLLHDLITSN SEQ ID MKGIEGVIMLSHDILPEKLLVSEKKHENVG NO: 74 SYFSDDIGEQSEQTEVSHFN SEQ ID MPNKKYSENTHQGKKPLIKSEANNEHAID NO: 75 NSPLGIGLDLNSILGNNSASL SEQ ID MEREYSEKEKHKKHPIQLRDAIEQHAEET NO: 76 ANNSLGLGLDLHQAINTPKVP SEQ ID MVHEYSINDRQKRHSFSSANPIDPEVTNR NO: 77 ENSRHRFPKDNYNKGHGDLFY SEQ ID MLKYANPQTVATQRTKNTAKKPPSSTSFD NO: 78 GHLELSNGENQPYEGHKIRKI SEQ ID MPRYANYQINPKQNIKNSHGKSSSSDFSS NO: 79 GYLSFSNNSLDDPFIRQQVKR SEQ ID MPNKKYSENTHQGKNPLMKSGANNEHDL NO: 80 QDSPLGIGLDLNSMLVNSSTSL SEQ ID MLSTEKHNKDTKHPRNREKKFSIQPENST NO: 81 QDDEDIKNNSLGVGLDLDQMI SEQ ID MPINDLKKKFEISPQAAQAIGAPARSNSSK NO: 82 QAEHQTEHLELDTSKNRRDR SEQ ID MPNSKYSEKVNHSANGAEKCSIHSNQYNI NO: 83 NNCTLGLGLDLNKKLRTGNER SEQ ID MPKLTELLSRFENPIQNQPNHISKKNPISN NO: 84 SKVLNNSEEKTAPLELKHDD SEQ ID MPRYSNSQRTPTQSTKNTRRTSPSSNSS NO: 85 TEHLSLSNAPTNDSSVRQEVKE SEQ ID MFSTYSSKNDNQTINKINTEEKHENTETD NO: 86 NHLEINLEHTGKSKPDIEPKD SEQ ID MKKTDEKYGQYEYKDEDITSYPIAWTNPD NO: 87 NGKIYIGINSPEYSHLNNKGE SEQ ID MVYEYDKTIERRRNPSIQLNNNEKSSEQA NO: 88 LELSQNNPLLHDLITSNNLRK SEQ ID MEHEYSEKEKPQKCPIQLRDSIEHDKEDIN NO: 89 TTTPLELNSQYTNRKRAGLR SEQ ID MYDSKKKNSEPTTKKKFERSNYSQWDDS NO: 90 INHYEDMNRARIKNRNDILTTV SEQ ID MPISNLAKESEVRAVKDIPCKNIETDNHLEI NO: 91 GLSSGLSRSKDTSKFKKNS SEQ ID MLANVLPNLASFLKYEKETPLFFIEDGFNF NO: 92 QNLNPGRVPLIKTPEQRKAG
TABLE-US-00006 (Photorhabdus asymbiotica strain ATCC43949 PVCPnf operon, pvc1 - pvc16; e.g. corresponding to genes PAU 03353 to PAU 03338 of the sequence of GenBank accession no. FM 162591.1) SEQ ID NO: 93 ATGTCTACAAGTACATCTCAAATTGCGGTTGAATATCCTATTCCTGTCTATCGCTTTATTGTTTCTGTCGGAGA TGAGAAAATTCCATTTAATAGTGTTTCAGGATTAGATATTAGTTATGACACCATTGAATACCGAGATGGTGTTG GTAATTGGTTCAAAATGCCGGGTCAGAGTCAGAGCACTAATATCACCTTGCGTAAAGGCGTTTTCCCGGGGA AAACAGAACTGTTTGATTGGATTAACTCTATTCAGCTTAATCAGGTAGAGAAAAAGGATATTACCATCAGTTTA ACTAATGATGCAGGTACCGAATTATTAATGACCTGGAATGTTTCTAATGCTTTTCCCACTTCATTGACTTCACC TTCATTTGATGCCACCAGTAATGATATTGCAGTACAGGAAATTACGCTGATGGCAGATCGGGTGATTATGCAG GCTGTTTGAAGCATTGATATTTAATCATCTCATATAAGGGAACTTTTATGACAACCGTTACCAGTTATCCTGGC GTTTATATTGAAGAATTAAATAGCCTGGCCTTGTCAGTTTCAAATAGCGCCACAGCGGTTCCTGTTTTTGCTGT GGACGAACAAAACCAATATATTAGTGAAGATAATGCAATCCGTATTAATTCGTGGATGGATTATCTTAATCTGA TTGGCAATTTTAATAATGAAGACAAATTAGATGTTTCTGTGCGTGCTTATTTTGCCAATGGAGGTGGATATTGT TATCTCGTCAAAACAACGAGTTTAGAAAAAATTATTCCAACCTTGGATGATGTAACCTTATTGGTTGCTGCGG GCGAAGATATTAAAACGACAGTAGATGTTTTATGTCAGCCAGGAAAAGGGTTATTCGCAGTCTTTGATGGCCC TGAAACAGAGTTGACTATCAACGGTGCGGAAGAGGCAAAACAAGCCTATACCGCCACACCATTCGCTGCGGT TTATTATCCTTGGTTGAAAGCGGATTGGGCTAACATAGATATTCCACCCAGTGCAGTGATGGCGGGAGTTTAT GCATCGGTGGATTTATCCCGTGGTGTATGGAAAGCGCCTGCCAATGTTGCGTTGAAAGGGGGCCTGGAACC TAAATTTTTAGTCACGGATGAATTGCAGGGTGAATATAACACTGGCCGCGCTATCAATATGATTCGTAATTTGA GTAACACAGGTACTACGGTTTGGGGTGCAAGAACCCTGGAAGATAAAGACAATTGGCGTTATGTTCCAGTGC GACGCTTGTTTAATTCTGTGGAGCGGGATATCAAGCGTGCCATGAGCTTTGCTATGTTCGAGCCTAATAATCA GCCTACTTGGGAGCGGGTACGGGCGGCGATTAGCAACTACCTTTATAGCCTGTGGCAACAGGGGGGATTAG UTGGCAGCAAAGAAGAAGACGCTTATTTTGTGCAAATTGGTAAAGGTATAACGATGACACAGGAGCAGATTG ATGCAGGGCAAATGATTGTTAAAGTCGGTTTGGCTGCTGTACGGCCTGCGGAATTTATCATTCTCCAGTTTAC GCAAGATGTAGAACAGCGTTAATCATATGATTATGAGGAGTTATCATGTCTGCTATTCTGAAAGCGCCTGGCG TTTATATTGAAGAAGACGCTTCCCTAGGGTTGTGTGTGAGTAAGAGGGGGAGTGGGGTGGGTGTTTTTATGGG AAAATTTACTCCGACAGTGGTTGATTCAATCCAAGTCTGTACCCGTATCAGCAACTGGCTTGAATTCACTTCC TGTTTTTGGGTAGGTGGAAGAGTTGAGATTGTTGTGGAATGTAAGAGTGAATGTGAATGTGAATGTGAAAGTTA GGAGTATATTGAGAGAATGAATTTATGTGGAGGTGTGGAAGGATTGGGAGTGTATTTTGAAAATGGGGGAGGA GCTTGCTATATCTACCCATTAAATGATGCTGAAGATGAATTGGTTCTGGCGGCCATACCAGAAGTCATTGAAC AGAAAGGTGATATTACTCTGTTGGTTTGCCCGGAACTCGATCTGGATTACAAAACTAAGATCTATGGCGCAGT GAGCTCACTGTTGAATGATAACAAAGTGGGCTATTTCCTGATTGCGGATAGCAATGATGGAGAATCTGTGTCA GGAGTATGGAATAGTGGTAAGGGGGGGGGGTATTATGGGGAGTTGGAAAGTAAGGTAAAATTTTGGAGGTTGG CTGGGGATAAGGACATTCGTATCAGCGGTTATCAGGATGATGATGAAACACATAAACCGAAAAACTTGGATG AGCTCAGGACAATCAACGAGGCGTTGGCACAGGATATTGATGCAAGATTGCTCGAGGAGAAACAACGTGCT GTCATCATTCCGCCAAGTGCTGCCATTGCGGGCATTTATTGCCAAACGGATAATCGTCGCGGTGTTTGGAAA GCGCCAGCCAACGTTGCGCTCACAGGGATCGGGAGTTTGCTTGATAAGGTAGACGATGAACGGCAGGGAGA GATGAATGAGAAGGGAATGAATGTGATGGGTTGATTTAGGGAGGGTGGTTTTATGGTGTGGGGAGGGGGTAG TTGTGTGGACGCTGCCAACATCAGCTGGCGTTATATTCCTGTTCGTCGCCTGTTCAATTCCGTTGAACGAGAT ATCCGCCAGGCGCTGCGCGCTGTGTTGTTTGAAACTAATAGTCAGCCTACCTGGGTACGTGCTAAGGCTGC GGTTGATGAATATGTTTATAGGGTTTGGGAGAAAAATGGATTGATGGGTGGTGGGGGGGAAGAAGGTTATTTT GTGCAAATTGGTCAGGATATCACCATGTCCGAGGCTGATATTAAACAGGGTAAGATGATCATGACTGTTGGTT TGGCAGCAGTGCGGCCAGCTGAGTTCATCATTCTGCAATTTACGCAGGATGTTGTTCAGTAATCTCCATGACT AAACGCCAGGCACTGTATTGACAGTGCCTACTCTAACCATCTTGGAGGAGGTGATGATGATGGAGAGACTCC AACCGGGTGTGACTTTAACAGAAAGTATAATCACGATGGGTCAGCAAGAGATACCCAGTGCTGTGCCGGTGT TTATTGGTTACACCGTTCGTTATCCGGAACAATCGGAAGCATCAGTCCGTATCGACAGTTTGGCCGAGTATAC GAGGGTGTTTGGTGAGGAGGATGTGATGATGTTTGGTGTGAGGGAGTATTTTGATAATGGGGGGGAAGAGGG ATTTGTTTTAGGGGTGAAGGAGAATATGGGATGAGTGGAGATGAGGAGAGGTGAAGGGGAAAATGTGATAGG GGGATTGGGGTGTGGTAGGGTTAGGGAAGGGATTGGTGGGGATAGTGAGATTAGAGTGATTTTGGTAGGGGA TATGGCTCGGCTTAATGACAGTGATATTGATGACTCCTCAACCCAGGTAAGCCTGTGGTCCCAAGGCTGGGA GGCGCTGCTGCAATTGAGTCAGGTTAGGCCCAACCTCTTTGTGCTGTTAGATGCGCCGGATAATGTTGAACA GGCGCAGAAGTGTATGACAACGCTATCGTCAGATTATCGTCAATGGGGGGCAGCATATTGGCCTCGTCTGG AAAGTAGGTATGAGAAAGAAATATGTGGGAAGGAGAATGAATGTGAGGGAATTTTGGAGGGGAGTGTTGTGTG ACCCACAGCCGCGGTCGCAGCGGTAATTCAACGCACGGATAACGACGCGGGTGTTTGGAAAGCACCGGCC AATATTGCCTTATCCCAGGTTATTCGACCTGTTAAATCTTATCTTCAGGGAAGTGTACTGTTTAACAGCAGCG GCACTTCGCTCAATGTGATCCGCAGTTTCCCAGGTAAGGGCATACGGGTATGGGGATGCCGCACTCTGGAA AACACGGATAATACGCAGTGGCGCTATCTGCAAACACGTCGGCTGGTTTCCTATGTAACAGCGCATTTGACC CAATTGGCTCGCATGTATGTCTTTGAGCCAAATAATGAACTTACCTGGATGAAGTTAAAAGGACAAAGTTACA ACTGGTTACGGCAATTATGGTTGCAGGGTGGCTTGTATGGTTCACAGGAGGATGAGGCATTTAACATTCTGT TAGGCGTAAACGAGACGATGACTGAGGATGATGTTCGTGCAGGAAAAATGATCATGAAAGTTGAGTTGGCTG TGTTGTTTCCTGCCGAATTTATTGAGATCAGTTTGGTGTTTAATACCCAAACAGAGGCGCTGTCTTAAGAAGG AAAAAGTAGGATGAAGGATTATTAGAGAGGGGTGGTATGGGATGGTTTTATGGGGAGTTTTATTTTTAAGGGGA TTGGGGATGGGGTGGATATTGGTTTTGAGGGTATGTGTGGGGTTAGTGGGGAAGTAGAGGTGAGTGAGTAGA GTGAGGGAGGAGAAAATGCCCGTAATAACTATTTAGCTGAGAAAATCCAACACGGTACGTTGACTTTGGAAC GGGGCGTGATGACAGTCTCGCCATTGACCTGGATGTTTGATCGGGTATTGAGTGGTGAAAAAATCGCTTATG CCGATGTGGTGGTGATGCTACTGAATGAAAATTCACTGCCATTGTCCAGTTGGACGTTGAGCAATGCGCTGC CGGTACGCTGGCAAACCAGCGACTTTGACGCTAACAGCAATGCCATATTGGTGAATACCCTTGAATTGCGTT ACCAGGATATGCGCTGGCTTGGAGTCAAAATATGACAGTAGAAATCAGAGAGTTACTTATCCAGGCAAAGGT AGTGCCATCAACACGACCGACTGAATCAGAACGGCAAAACCATTCTTTGATACAGGAAAGTCTGGATGAGGC GACTTGGGTGGAAACGATAAAACGCGAAGTGTTGGCCGCATTACGCGATGAGGAAGGGTGGCGTCCATGAG TCTGATTGAACGTGGTTTAGCTAAGCTGACAATTAATGCTTATAAGGATAGGGAAGGGAAGATACGGGCAGG AACGTTGCAGGCCATGTATAACCCTGACTCCTTGCAACTGGATTACCAAACGGATTATCAGCAATCCCAAGC GATTAATAGGGAAAAGGAAAGTAGGATTTATGTAGAGGGGAAGGGGGGAGGGTTATGAGTTGAATTAATTTTT GATGCCACGATGCCGGGTAACAAAACCCCCATTGAAGAGCAGCTCATGCAGCTCAAGCAACTGTGCAGTGT GGATGCAACCAGTAACGAGACGCGATTCCTGCAAGTTAAATGGGGCAAAATGCGTTGGGAAAGTCGGGGTT ACTTTGCTGGCAGGGCCAAGAGTTTGTCTGTGAATTACACTTTGTTTGATCGTGATGCGACTCCCTTGAGGGT ACGGGTAATATTGGCATTAGTGGCTGATGAAAGTCTGGTGTTGCAGGAGACTGAACAAAATCTGCAATCTCC GGCAAAAATCGCATTACGCATACAGGATGGGGTATCTCTGGCTCTGATGGCAGCCAGTACGGCATCAACATT GTCAGGCGGTGTGGATTATCTGACGCTGGCCTGGCAAAACGGTCTGGATAATCTCAATGGGTTCGTTCCGG GTGAAATATTGCAGGCCACCAGGGGAGACGAATCATGAGCCACCAACTGAAAATTATTGCAGATGGTAAGGC ACTGTCACTTTTGGCCGCGGTAGATGTGGACACCTGTTATCGGGTTAACAGTATACCTTCTGCGACATTGAAA CTGAGCGTACCGGATAGGCCACTCTCTTCTTTCAGTCAGACGGATGTTCAGACAGAACTGGCCCACTGTCAG GTAGGGAAAACCCTGCGTCTGGAATTGATTGATGGTAGCAAAAAATGGGTGCTGTTTAATGGTCTTATTACCC GTAAGGCTCTGAGAATTAAGAATAAGCAATTATTGCTCACTCTGGTTGTCAAGCATCGGTTGCAACTGATGGT GGATACCCAGCATTCACAGCTGTTTAAAGACAAAAGCGAAAAAGCGATCTTAAGCACGCTATTGAATCAGACC GGAATCAATGCTCGCTTCGGAAAGATAGCGGCGTTAGATCAAAAGCATGAACAGATGGTGCAATTTCGTTGT TCAGACTGGCATTTTCTGTTGTGCCGACTGTCGGCAACCGGTGCATGGTTGTTACCTGCCATAGAAGACGTT CAGTTTGTTCAACCTGATGCTCTGAAATCAAACTCAGCCTATACCTTGAAGAGCAGGGGGGATGAGAACAAA GACATCGTTGTCAAGGATGCTTACTGGCAGTTTGACAATCAAATCAACCCCGCTTTGCTGGAAGTCAGTGGC TGGGATATCAGTAAGCAGCAGGTACAATCAGGCGGTCGCTACGGAAAAATCGCGTTGGGTAAGGCGGCACT CTCTCCTGATGGATTGGCATCCCTTAATAAAACGGGTTGGGACATTTGTTATAGCAGTCCGTTAACAACCCAG GAAAGCGGTTATCTGGCACAGGGATTATTGCTTAACCAGCGCATTTCTGGGGTGACAGGAGAATTTTGCTC AAAGGAGATGGGCGTTACCAGTTGGGAGACAACATTCAGCTGACTGGATTTGGTTCACAGTTAGATGGTACG GCAAGCATTACTGAGGTTCGCCACCGTCTTAATCGGCGAATTGATTGGGAAACCACGGTGAGCATTGGTTTA CAACATGAATATTTGCCGATATTACCTGATGCTCCCGAACTACATATTGCGACAGTAGCGAAATATCAGCAGG ACAGTGCGGTGTTAAACCGTATCCCCATTATTCTGCCGGTACTGAATCGTCCCAATGAATTTTTGTGGGCCAG ATTGGGGAAACCTTATGCTAGCCATGAAAGCGGTTTCTGTTTTTACCCAGAGCCAGGTGACGAAGTTATTATT GGTTTTTTTGAAAATGATCCGCGTTATCCAGTTATTTTAGGTGCTATGCATAATCCGAAAAATAAGGCCCCTTT TGAACCAACCCAAGATAATAGGGAAAAAGTATTGATCGTTAAAAAAGGTGAAGCGCAACAACAATTAGTCATT GATGGCAAAGAGAAAATGATCCGAATTAATGCGGGTGAAAATCAAATAATGCTTCAGCAAGATAAAGACATTT CTCTGTCAACGAAAAAAGAATTAACACTGAAAGCGCAGACAATGAATGCCACGATGGATAAATCATTGGCAAT GTCCGGGAAAAACAGTGTTGAAATCAAAGGCGCAAAAATTAATCTTACCCAATGAAAGGTGACGATGAATGG AAAATCAAATACTGACACAACTCTATGGTCGTGGTTGGGCTTTTCCTCCGGTCTTTTCCCTTGAAAAGGGGGT AGAGATGGCTGAAGGGGCGGAAGATGTGAGACAAAGTTTGCAGATTCTGTTTAGTACTGAGCCGGGGGAAC GTCTTATGCGTGAAAATTATGGCTGCGGATTAAATGATTTTATGTTTGAAAATATCCGCAATGAACTTATTGCT GAAATTGAATCCCATATCCATGACAACGTATTACGATATGAACCCCGGGCTGATATGACTGATATTCAGGTTC GTCAATCCCCTGGCATGGGGAATACTTTGCAAGTGCAGGTCATGTATCGCCTGAGAGGGAGTGATATCAATC AACAAATCCAGGGAGTACTTGCACTGAGTGAAGGCCGGGTGACGGAGGTAGTATGAGTGAAGCGATTGTGG TGGATGGTGACGTGTTACAGTTTGATCCCAACTTTGGCAATCGGCAGGTGACGGTTCCCAGCCCAGGAAAAA TTAGCGGCACAGGACATGCGCAGGTAAGTGGAAAAAAAGTGTGTATTCTGGGGGATGAGAAACAGGTCAGG GTTTCTGCAACCTATATTACAACAACACATACTACGCCGGGAACAGGAACCATTACTATCAGTGCTCTGGATG CTGGCCAGCAGGCCCTTCAGTGTACCAGTGGGGCGGCTTTAATTATCAAGGGGCAGCAATTTACGGCGATG TTTACGCCTGAATTGCCAGCCATGAATAATACAGTGACTCCGCCACAACCGGATGTTACGACACCTTCATCAG GAAAAGGACGTTTTATCACTCAACAAAATTTTGCTACCGTAAATTAGAGTATTGACTGAATTAAATAGAATTAA CGAAGGTGTAAATAATTATTTATTTGCTGACGAATCGCTGTGACAAATAAACACAGGTGATGTTATGGAATTAA ATGAGTTAACTAACAAATTGTCAAATTTGGTGCCAATGACCGATTTTAAATTAGATAATCGAGCCAGTTTGCAA TTGCTTAAATATATTGAAGCGTATACGAAGATAATACCCTTTAATTCTGGCGATAAATATTGGAATGACTTTTTC TTTATGTCAGGAAATACGCCAGAGAAACTTGCAAAATTATATCAGAAAGAAATAGAACCCAATGGGGAGTTAT TACCTCAGCAGGCTTTTTTGTTGGCGGTTTTGCGTTTATTGGAAACACCAATATCCTTATTAAATGTATTACCT GCTGCTCATCGTGAGCTCTATTATCGGGAGCTTTTAGGCTTGTCTTCCCATGCGGCACAGCCTGATCAGGTT GCTTTATCTATGGAACTGAATTCGACAGTGATGGAACAGCTGCTCCCTGAAGGAACCCTGTTTGAGGCTGGT CAGGATGAACAAGGCAATGCATTGCAATATGCCCTGGATGCCAGTTTGCTGGCTAATCGTGGATATATCAGT GACTTGCGCTGGTTACGGAATGACGGGGAAAAGCAATGGGTTACTTCTGCTCCATGGGATTTACAGGCACAG GTGTCACTGCCGTCTGATGGGATACGATTATTTGGTAAGACAAATAGTGATCAGCAGGTATTTGGTGGGGTG TTGATAACGTCATCACTTCTGGCGATGGAAGCGGGGATAAGGAAGATCATTGTTACTTTTGAGCAGGAGATG AACACCCAAGAACTGGTGGCACAGGTCAGCAGTGGAAATCAATGGCTAACATTGACGTCTGAGGTAAATAAG AAAGAGGTCACACTGACACTGTCAGACAAAGAACCGGCAATCAGTGCGCCAGAGGATCTGGATAATCTCTTT TTCACGCAACCGGTACTCAGGCTACAGGGAAAGGATAGTCAGGCACTGCCGGAGGTGACGGGTATCAGCGT TTCGGAAAAGGATGATACTAAGGATACCTCTTTTGAGATGTATCACTTAACACCATTTGGTTATAGCAGTGATA TAGAGCCATTGGAGGAAAATCCAGCGTTATATTTAGGCTTTACTGATGTAAAGCCAGGGCAAACACTGGCGC TGTATTGGAAATTAAAATCCCCGCAGCAACCAACCGTTTCCTGGTATTACCTGGATCAACATAATCAATGGGC TGAATTGGATTCATGGGTCAGTGATGGAACCCAGAATCTGTATCAGGATGGTACTTGGCACGTTGAGTTGCC TGTGGATGCATCCAATCAGGCAGAGCAGATGCCAGTTGGACGCTATTGGTTGCGGGCAGTGGTGGAGGTAC CCGCTCATGAGGGGGCGTTGGGGAAGGCTCCTTGGCTATATGGTCTAATCTATAACGCCATGACGGCAACC TTGGTTAATGTAGATAGCATCAGTGACAGCCATTTCTTAACCCCTTTGCCTGCCAGCAGCATACAGCGGCCC GTTGAACCCATCATTGTGTTGGCATCGGTCAACCAGCCTTGGGCATCATGGGGTGGACGTATACCTGAATCC TACAGTGCCTTTTTTGAACGGATAGCTCAAAACCTGTCTCATCGAAACCGGTCCTTAACCTGGGGAAATATGG TGACATTACTCAAAGAGCGTTATGTCAGCATCTTTGATGTTAAGTATCCAGGTAATGATGAACTCACCAGAGT GCCAGCATTGGAGCAGCAGCAACTAACAGTGATTCCAGCAAACCGGTACAACGATAGCGATGATTCTCTGCG TCCGGTACTGAATCCTGCTCGTCTGCAAGAGATGGCTGATTGGTTGCAGCAGAAAGACTCTCCCTGGGCCTC TATTGAGGTCAGGAATCCAGAATACTTGGATGTGAAAATCCATTACGAGGTGATTTTTAAACCTGATGTGAAC GAAGATTTTGGCTATCGCCAGCTACAGCAGCAACTGTGTGAGGTGTATATGCCTTGGAGCATAGATGAGCAG CGGCCCGTTGTATTGAATAACAGCATTAATTATTTCCAGTTGTTAGCCACTATTCAACAGCAACCGCTGGTTG AGCGAGTCACTCGTCTGACACTACATCGGGCTGATTCTTCTGATGAGAGTGATGGTACAGCATCTGTGGAAG CCAAAGATAATGAAGTGCTTATTTTAGTCTGGGAAGAGGACGATAATCTGCAATACCGAGGAAATGACTATGA GTAATCAGGATGCACTGTTTCATAGCGTTAAAGACGATATTCACTTTGATACCTTGCTGGAACAAGCTCATCA GGGATTGAAAAACAGGCTGAAAAACTGTGGAGTGATACGGCAGAGCATGATCCGGGTATGAGATTTTTGGA GGGAATCAGTTACGGTGTGTCAGATTTGGCTTACCGACATACATTACCCCTGAAAGATTTACTGACTCCGGC GGGGGATGAGGAGGAGGAAGAGGGAATTTTTGGTGGGGAATTTGGGGGGGATAATAGAGTGAGTTGTGGGG CGGTGACAGCGGATGATTATCGCAAGGCATTGTTAGATCTACACAGCAGCGACAGCCTGGATGGTACTCAG GAGGATGAGGGGGATTTTGTGTTGGGGAGTGTGGAAGTGGTGGGTGAAGGGGAAAAAGAGGGTTATAGGTAT TGGTATGATGCAACCAAGAGGGAATATAGCTTTGTCAACAGTGAAGGGGCTAAAGAGTTTACCTTGCGGGGG AATTACTGGTTGTATCTGGAACCAACCCGTTGGACTCAGGGTAATATTGCCGCTGCTACCAGACAACTGACA GAATTTTTGAGTAAAAATGGGAATATTGGTGAATGTGTGAGGAAGATTATGTGGGTAGAAGGGGTTGATGTGG GAGTGTTGGTGGATGTTGAAGTGGATGATGATGTAGGTGGAGAGGATGTGGGGGGTATTTTTGGGGGGGTGT ATAGCACCGCAGAGCAGTATCTGATGCCTGGAGCACAGCGTTACCGTACGGAAGTACTGCAAAATGCTGGG ATGAGCAATGATCAAATCTTCGAAGGTCCATTATTGGAACATGGCTGGATACCAGAGCTGCCGGCAGCCCGT GATTATACTCAAAGGCTCACTCTCAATCTTAGCCGGTTGGTAAATAGTCTGCTTGAGATTGAGGGCATTAAAC ATGTGAATCGTCTTCGTCTGGATGATAGCTTCGATAAAACTGCTATTGAACCCGTTAAGGGGGATACCTGGTC GTGGTCGATCAAAGAGGGCTATTATCCACGTCTTTGGGGAGAAGACCCACTTAACCAATTGGCGCAACAAAA TGGCCCGCTTAGGGTGATAGCCAAAGGAGGGATTAGCGTCAGTGTGAGTAAAGAGCAAATCCAGGCCAGTT TACCCAGTCAATCACTGATTCAAAATGAGCCGGTAATATTGGCTTACGGCCAGCACCGTGACGTTGGCAGCT ATTATCCCGTCAGTGATACTTTGCCGCCTTGCTATGGACTACAACATTCTTTGTCTGAAAGTGAACACTTATTG CCACTTCATCAATTTATGTTGCCATTTGAACAATTATTGGCCTGTGGTTGTCAACAGATAGCCATGCTCCCGC GGTTACTGGCTTTTCAGCGCGAAGGTTATGAGGTTTGGGGTGATCAGTGGCCCTTTAAGTCAGGCTCAGTGA ATGATGACGCCCATCAAGATTATGCCCCTGCATTAAAGGATTTGTTAGGACAGATTGCGCTGGATAGTGATCA TGAATTGGATATTATTAATTACTTGCTGGGTTACTTTGGCACACAGCGGGCACCGCGTACCTTTACGACACAA CTUGATGATTTTCGTGCGGTCCAACAGGGTTATCTGGCCCAGCAACCGACATTGACTTACCACCGCTCCAAT ATTCGTATCGATCAGGTATCGTCGCTACAAAAACGTATTGCTGCTCGCATGGGGCTGGGCGGTGAGTTGTTT AAACCTCAACCGGATCTGAGCCAACTGCCTTTTTATTTGATTGAACATCGAGCGTTGCTGCCAGTCAAACCCA ATAGTCAGTTTGATAAGGAACAGAAACCAGCCTCGGTGACAGAGGAGGGGGGCAGCCAAACAGGTCAACAT TATGTGGTCATTGAACAGAAGGGCATTGATGGCAAGCTGACACAGGGGCAAGTGATCAATTTAATTCTGTAT GAAGGAGAGCAGGGAGAAACCCAATTTACGATACGCGGTCAGATGGTATTCAAAACCGAGGGGGATAAGTT TTGGTTGGATGTGAATAATAGTGCGCAACTGGAATATAATCTGGCGCGGGTAATGACAGCAGCCAAGGCGAG TAAACTCTTTTGGCAAAACAGCCCGGTATGGATGGAGGATATGGGCTATCGTCTGGCCTATGCTAGTGACCA ATCCTCATTGCCTGTGAATCAACGGCGCTTGACCCGCACAGTGCAAACTCCATTCCCGCCGATGGTTGTTGT AGGTAGCGAAATCACCCTGTTAAAGCAGGTGGGGATAGTCAATTTAAAAAAAGCGGAGTCAGAAAAACTTTAT GCAAAAGTTGTTAGCTTTGATCGCATTGAAGGGACCTTGATTATTGAGCGTTTGGGTAATTCCACTCTGGCTT TTCCTACCTCGGAAGAGGCGTGGCGGTATAGTTGGTATTTTTCGGGGGAGAAATATGAAAGGACTGACCGCT TTTCATTTGTGATTAGCGTAGTAGTGAACAGTGACTTAATTAAATTGCCCGGTGTTGATCCCTATAAATTGGAA GAATGGGTGAAAGAAACGATTCTTACCGAATTTCCAGCTCATATTTCTATGATTATCCATTGGATGGATCGGG AAGCCTTTTTAAATTTCGCCAATACCTATCAGCGTTGGCAAAATAATGGTACGCCACTGGGGGATGCGGCTTA TTCCATTCTAGAAAGTTTGACACTTGGTAAATTGCCATCTGCCTTAAAAGGTGTTGGCACAATGCGTATTGCC ACATCTAGTCAAAGAGAAGAAGTGGTGGGTAGTAATGGTGATCAATGGAATACAGATGGAATAACCCAGAAT GAATTATTCTATGTTCCTAAAGAGAGCTAGGAAAAATAAATATCTGCCACTAATGATGTTGAATTAAATATGTTT TCTGGAGTTAATCATGAACGAAACTCGTTATAATGCAACTGTACAAGAACAACAAACATTATCTAATCCAAAAG CTGTTGGACCTGACATCGATAAATTAAAGGATAAATTTAAAGAGGGCAGTATTCCCCTGCAAACCGATTTCAA TGAGTTAATTGATATTGCCGATATTGGACGTAAAGCCTGTGGTCAAGCGCCACAACAAAATGGCCCAGGAGA AGGATTGAAATTGGCTGATGACGGTACGCTTAATTTAAAAATAGGCACTTTTTCCAATAAAGACTTTTCTCCAT TAATATTAAAAGATGATGTTTTATCTGTAGATCTTGGTAGTGGTCTGACTAATGAAACCAATGGAATCTGTGTC GGTCAGGGCGATGGTATTACAGTTAACACTAGCAATGTAGCTGTAAAACAAGGTAACGGAATTAGCGTTACTA GTAGTGGTGGTGTTGCCGTTAAAGTTAGTGCTAATAAGGGACTTAGCGTTGATAGTAGTGGTGTTGCAGTTAA AGTTAATACTGATAAGGGAATTAGCGTTGATGGTAATGGTGTTGCAGTTAAAGTTAATACTAGTAAAGGAATTA GCGTTGATAATACAGGTGTTGCAGTTATAGCTAATGCTAGTAAGGGAATTAGCGTTGATGGTAGTGGTGTTGC AGTTATAGCTAATACTAGTAAAGGAATTAGCGTTGATGGTAGTGGTGTTGCAGTTATAGCTAATACTAGTAAA GGAATTAGCGTTGATAATACAGGTGTTGCAGTTATAGCTAATGCTAGTAAGGGAATTAGCGTTGATGGTAGTG GTGTTGCAGTTATAGCTAATACTAGTAAAGGAATTAGCGTTGATGGTAGTGGTGTTGCAGTTATAGCTAATAC TAGTAAAGGAATTAGCGTTGATAGTAGTGGTGTTGCAGTTAAAGTTAAAGCTAATGGCGGAATTAAAGTAGAT GCTAATGGTGTTGCAATTGATCCTAATAATGTACTCCCCAAGGGAGTGATTGTAATGTTCTCTGGCAGTACTG CACCAACTGGTTGGGCGTTATGTGATGGCAATAATGGTACACCAAATTTAATCGATCGATTTATTTTAGGTGG GAAAGGGACTGATATTAATGGAGTGAGTACTAATACAGCTTCAGGTACTAAAAATAGTAAGTTATTCGATTTCA GTTCTGATGAAGCTACATTAACTATTGATGGTAAAACACTGGGGAGAGCATTATCGTTACAGCAAATACCTAA TCATGCACACTTTAGTGGAATAATTATGGATACAGAGAAAGTTAATTATTATGGAAGTAAAAAAATCACAACAA ATGTGTGGGGTGTAACAACAGGAGATAATACTTCAGTACGATATATTTATAAGTCATCAGGTGTACTTGACTC TAACAATAATGTCTCCAACAGTACCTTAGGCGGAAACAGTCTGCAGACGCACGATCATGATATTAAGATAACG GGCACAGGAAAACATTCTCACAAAAACAAAGTAACAGTCCCTTATTATATTCTGGCTTTCATCATAAAGCTTTA ATATATATGAAAAATTGAAAATATAAATTATCCATTAATAATAAAGAGGATATTAGCATGACTTCGGAGCCAAAT CTGTTAAACCGGATTACAATTACTATTGAAGCTAATAATCAACAAGTAGCTAGAAAAGTATTGCATGGCTCCTT GCTTAATCAAGCTAATATAAATAAATTATTTAATTCATACTTTAATGAATATGAAATTAATAGGGGTGTTTATTTA GAAACATTAATCCTGAATCTTGGTACGATAAATTTCCATGATTTTAATTCATTGTTTCCTACTCTCCTAAAAGCT GCATTGAATAAAGAATTCAGTCAATATCAGATAAACAACCATAGGGAAGAAATGCTATTTAATGAGACAATATC AAATCAAGCTACTGATAAGTCTTACATATTTGGCGATAACAAATTAATTGATGCAGAGAATTTCATTCACTTTTT ATATCAAAAGCATTCCACATTAAATCTAGTAGAAGCAATGGGAAATAATGGTATTGAAAAATTAACAAATCAGT TAACACAAATAGAAAATAAATTTGCGTTATTATTGGCAAAAAGTTGTTTGTCTGAGGAAGGCTTAAAACGACTC TTGGCTATCAAACAACCCGATTTATTAATCGCTATCAATCGCAGATTATCTGAAAGAATAAATAGACCACAATA TCAGGAGAAGCTTGTTTCCTGCGGACAACTGATATTTAGTGCTCTGGGATATATACAACAGTACAATATACAG GAAATTCCTAAACCGGATGAAAAAGTTATTGCACGCATAACAACTGAACTTAATAATAATGGTTTGCTTAATAC AATACCTATTATTACACTATTTCGTCAGAGTGGGATTAACGATTCATCACTAAATGATTGGCTAAAGAAAATCT GGCAGGTGAGATCAATTTCACAGTTATGCAGAAAGTATCTTTCTGCTAAGGAATACCAATATCTGTCAGAACA TTTTGTTTCAAAGAGCGTCGATAAAAATAGATATGATGAAGAGCCCGTAAATCAGAGCATATTATCAAGGTTG AATAATAATTCCATTAAAGAAGGAAATAATCACAGTCAACTCTGTACTCTCAGTAGACTATATTCTGAACCCGT TGTATTACCTGAACAAACCATTCTACGTCAGGTTAGTAATACAGTAGATCAGAGCATATTATCAAGGTTGAATA ATGCCTCCATTAAAGAAGGAAATAACCAAAGTCAACTTCGCACTCTCAGTAGACTATATTCTGAGCCCGTTGC ATTACCTGAACAAACCATTCCACGTCAGGTTAGTAATACAGGTATATTAATTCTATGGCCAATGCTACCTACAC TATTTAACCAGCTTGGTCTACTTGAGAAAAAGAAATTTATCCATCGTCAGGCCCAGTTTAATGCCGTTGATTTT CTTGATTACCTGATTTGGGGAACCGAAGATGTGAAAGTGGAACGAAAGGTTTTGAATAATGTTCTATGTGGGT TAATGGCTGATGAAATTACTGAACCAATGCCTATTGAACCAGAAAAACAATGGATAATAATTCAATGGCTGGA CGCTATTATCTCCCAACTTTCTGGCTGGAAAAAGTTAAGTCGTAATGACGTCCGTCAATTATTTCTACAACGAC CAGGAGAATTACTGATCAATGAACAGGAAATTAAAATCACAATACAGCAACAACCATTTGATGCTCTGTTAACT GATTGGCCGTGGCCAATGAATATGGCTTGTTTTAGCTGGTTGAGTCAACCATTAACCATTAUGTGGTTATAAC GATTGAGGAGAATGAGTTAGTGTGAGTAAAAAATATGAATATATGGGGTGTTTTTTATGATTGATTGAATGAGG ATAAGGAGGGTGATGTATGGTTTTTATTTAGGGAAGTGGAAGGAATAGATGTGGGTGTTGAAGAGGATTTTTAT TGTGTAGAAAGTCAGCGAAGTGAGCTCCTGGATGAGTTTCTGCTCACTGAGGCGGAAGTGGTGACCAGGCT GGATAAGCCACTTGGTAAACCTCATTGGATAAATGATGATTATCTGGCGATATCGCAAAAGGGCAATGTAAGC GTAATGGGAGGGTGGAGATTAATGGATGTGATGGAAGGGTTTGAAGTGAGTGATTTTGAGGGGGATGTTTTAG TATTAGGGTTATTGGGGGATTTTGATAGGGGGTATTATGGAGTGTTTTGGGTGATTGAAGGGGGAGAAGAGGG TGGATTAGGTTGTTTTGGGGTGGGATTGGAAGTGTTTTGGGAGTGGGGGGTGGAGAAAGAGGTAGAGGAAGG GAGTTTTGTGGAGGGGGGAGGTTTGATGGGTTGGGAGGTATTATGGATGGATAGTAGTGAAAAAAGGGTGGG GTGGGTGGAGAGTGGGTTTATTAGTGAGAGGGGGGTATATGAGTTTTTAGTGGGGGATGAGTAGATTATGGGG GCTTTAGAACATTGTGCTGAGTGGTTAACACCGACAGGGATTGGCTGTTATCCTGAAGGATTAAAACAAGTAC TGGGTAACGTATTGTTATCTGACAACGATAATATTAGACCGATTGTCTTATTACGGGGAATGGCCGGCAGTGC CAGAGCTTATACCATTACTAATATGATGGCTTCAGAAGGGAAGCAAACACTGCTGGTAGATATATCCAAACTT GGTGATAGGGATGAAAAAAAGATTATTGTTGAGATAAAGGATATTTTGGGGGAAAGGGGGATGGATGGAGGAT GTTTATTATTAGGGAATTTTTGGTTGTTAGTGGAAGAGAATAAAGAAGTATTGGAGTGGGTGTGAGAGTTATTG AATCAACCTGAATTAAGAATTGTTTGCCTGATTGAGCCTTATTCCCCATTGGTATGGCTGAAAAAGATACCGG TATTAGTGATTGAGATGGGAGTTTTAAGGGGTGGGGAAAAAGGGAGATTGTTAATTGGGAGGTTAGGGGATAA TTGTTGGGAGGATATTGATAGGATAAGTTTAAGGGAGGGTTAGAGTTTTAAGGGAGAAAGGGTGGGATTGATTT TGCAAGAGGCCCAGCTTTATCAACAGCAGCGAGATCCGCTGGATATCTTGCAGCAATGCGATATACGCCAGG CATTAAATTTGCGTGCTCAACAAAATTTCGGTCAATTGGCACAGCGGATTATTCCTAAGCGCTCATTAAAGGA TTTATTGGTATCCGATGAGATTGCTCAGCAGTTACGGGAAATACTCATAGCAATTAAGTATCGGGAACAGGTT GTGGGGGGAGGGTTTAAAGATAAAATTGGGTATGGGAGTGGTATGAGGGGGGTGTTTTATGGTGATTGAGGG ACTGGAAAAACCATGGCAGCAGAAGTGATTGCTGACCACATTGGCGTTGACTTAATAAAAGTGGATTTATCTA GAGTAGTGAATAAATAGATGGGTGAAAGAGAAAAAAAGTTATGGGGTATTTTGGATTTGGGGGAAGAGGATGG AGGGGTATTATTCTTTGATGAAGCTGACGCACTGTTTGGTAAACGCAGTGAAACTAAAGATTCCCAGGACAGA GATGGGAATATTGAAGTTTGTTAGTTATTAGAGGGGGTGGAGAATTAGGGGGGTGTGGTGATTTTATGGAGGA ATAATGGTGGTGATTTAGAGAGTGGTTTTAATGGTGGTTTTAGTTTGATTAGGGGTTTTAGTTAGGGGGATGAA AAAATGGGTAAAAAAATGTGGGAGGAAATTTGGGGTAGAAATATAAAAATATGGGAAGATATGGATTTTAAGGA ATTAGCTCAACGAACAAGCGTGACTGGCGCGAATATCCGCAATATTGCTTTATTGTCTTCATTCTTTGCTTCA GAGCAGGGGAATGATGAAGTCAGTAATGAAAATATTGAAATTGCATTGAAGCGTGAATTAGCTAAAGTCGGA GGATTAAGATTTTAAAAGTTATGAGAATGAAAGTATTGAAATATTAAATAAATTTATTAGGAAAAAGTTATGAGG ATATAATTTAAGAGAGGTTTTTTATGTTAAAGAGGGAAAGTATTATTGATGTGAATAAGGGAATGGATGGGATG GTGGGGGGATATGTGAATGAAGATATTGGGATTGGTTTTGATGTAGGTGAATTGGATAGTATGGAATGTGATGG GATGGTAAGTATCTTTCTTTATGACATTCATGAAGATTTACAGCTTCGCTCGGCAGAATCAAGAGGGTTTGAT GTTTATGCCGGGAGGTTATTGCCTGGTTGGGTAAATATTAAATGTAACTATCTGATTACCTATTGGGAAGCTT CTAAGCCAGCGACTGATGCCAGCAGTCCGGATAGCCAACCTGATAACCAGGCAATACAAGTGATGTCACAAG TATTAAATGCCTTGATTAATAATCGTCAATTGGCAGGTATTCCTGGTGCTTATACTCAGGTTGTACCGCCTAAA GAGAGTTTAAATAGCCTGGGGAATTTCTGGCAATCACTGGGTAATCGCCCACGGCTTTCTCTCAATTATTCAG TGACAGTACCTGTTAGCCTAAACGATGGTCAGGATAGCGCGACTCCGGTTACCGCGGTTTCTTCTACAGTGG AACAAACGGCATCGCTCAGTCAAGAAGTGGTTAGTCATGCTTTACGCGAATTACTCATTACGGAATTAGGAG GAGGAGAGGATAACCGGTTGGTACTGAGTAAAGTTGAATTATCCGCAGTGAAAGAGACGATGACTCAAGACA GTCCGGCTCAGATGATTATATTGTTGTCTGTTTCAGGCATTACACGACAGGAATATTTGAAGGAAATTGATAAT ATCTTTGATCGTTGGGTAAATAATGCTGAAGTTATTACCACTATTGATGATTGTGGGATTAGAATTGAAAGTAT AACGAAAGATAATCTTGTAGGAATTTAA (Photorhabdus asymbiotica strain ATCC43949 PVCIopT operon, pvc1 - pvc16; e.g. correspondinq to genes PAU 02112 to PAU 02099 of the sequence of GenBank accession no. FM 162591.1) SEQ ID NO: 94 ATGGCCACAACCACAGTTGACTATCCAATACCGGCTTATCGATTTGTTGTCTCCGTTGGTGATGAACAAATCC CTTTTAACAGCGTTTCGGGGCTGGATATTACTTATGATGTCATCGAGTATAAAGATGGCACCGGTAATTATTAT AAAATGCCGGGTCAACGTCAGTTAATCAATATTACACTGCGTAAAGGGGTATTCCCTGGCGACACTAAACTTT TTGATTGGCTTAATTCCATTCAGCTTAATCAGGTTGAGAAAAAAGATGTTTCAATTAGCTTGACCAACGAAGTT GGAACTGAAATTTTAATGACCTGGAGCGTAGCCAATGCATTCCCAACCTCATTAACATCTCCTTCTTTTGATG CCACCAGCAATGATATCGCTGTTCAAGAAATAAAACTGACTGCCGATCGAGTCACTATTCAGGCAGCTTAAAG CATCACGATGATTGATATATCAGACGGGACAAAATGATCCTCAAAATTTGGCACAACGGCTACCCGTCCAACT AAATTTACCCTCTTACAGTTCACGCAAAATATCGCACAATACAATTGGAGGCAATATGCCAACAACAACTTATC CCGGCGTTTATATTGAAGAAGACGCCTCACTGTCACTTTCCGTTCGCTCAAGTGCAACGGCGGTGCCCGTTT TTACCGTTGAAGATGACAGTCAACTTCATACTCCTACCAGAGTGAATAGTTGGTTAGAATATCTGACAAAAAAA GCAGATAAAAAATTCAATTCTACCGACAAACTTGATATCGCATTGCGCGCTTATTTTATTAACGGCGGCGGAT ATGGTTATCTCGTCAAAGCGGGTGAATTAACAAATCAAATTCCAAAACTTAACGATGTCACATTACTGGTCGC GGCTGGAGAAAATATCAAAGATGCTGTGAGTACACTTTGTCAACCGGGCAAAGGCTTATTTGCCATTCTGGAT GGCCCAACCGAAGAGTTAAAGTCTGATGGCAAATCCAGAGATCCGTATGATCAAAGCCCTTTTGCCGCCGTT TATTACCCCTGGCTAGTTGCTGATTGGGCAGACAATATTCCGCCAAGCGCGGCCATTGCCGGTATCTATTGT TCAGTTGACCGTACCCGCGGTGTCTGGAAAGCCCCAGCAAATGTCATATTACAAGGCGGGGTGAAACCGAA GTTTAAAGTCACCGATGACTTACAAGGTATTTACAACACCGGTAAAGCCATCAATATGATCCGTGAATTTCCG AATACCGGTGTCACCATCTGGGGCGCCCGCACACTTAAGGACGAAGATAACTGGCGTTACATCCCAGTTCG CCGCCTGTTTAACAGTGCAGAGCGAGACATTAAAAATGCCATGAGTTTCGCGGTCTTTGAACCTAACAGCCA ACCCACCTGGAAAGCTGTACACCGAGCTATTGATAATTATCTCTATGCCCTTTGGCAACAAGGAGGGCTAGC AGGAAACAAAGCTGAACAAGCTTACTTTGTGCAAATTGGTAAAGGGATAACCATGACCGATGATGATATCAAG CAAGGGAAAATGATTGTTAAAGTGGGTATGGCCGCAGTGCGCCCGGCTGAATTTATCATCCTTCAATTTTCAC AAAATGTAGCACAGTAACCGTACTGAGGCGCGGTTTAACACCGCGTCCATTCAGTCTATTGAATGGAGGAGA CAATAATGATAACGGAGATAAAACAGCCGGGCGTCACCATCACGGAAAATTCGATATCCCCGAAATCAGATA ATGAATTTATCGGCGTCCCCGTTTTTATTGGCCATACCGAAAAAAATTCAAGCCATAAAACGGCTGTTAAACTA AATAGCCTGATGGACTTTACCCAAGCTTTCGGTGCATCAGGATTAACCTATTATTCAGTACGCCACTTTTTTGA AAATGGTGGACAGCAAGCTTATATCTTGTCACTGGGGATTAATCAACAGCTAAAAGATTTTCAATCATTGATTA CCGCCCTGCAATGGAACTGGGTAAAACAAGCCATTGCCGCAGAAAACGAAATCACATTGATTGTTGTGCCTG ATATTACCCGTTTTAATGATCTCAGCGCTCAAAAAAGCCTTTGGCTACAACTCTGGCAATCAATACTTGAACTG TGTAAAAGTCGGCGTGGCATCATGGGATTACTGGACGCGCCTGATGATCCAACATTAGCAACTGAGTGTTTA AAACAATTCTCTTCCACTGATCGCCAATGGGGCGCCGTATACTGGCCAAGGCTAAAAAGTACCTACCAAGAA AACGGTACATACATTGTACTTTCACCTACTGCTGCGGTCGCCGCCGTTATGCAACGCAATGACAGTCAGAAA GGCATATGGACTGCTCCCGCCAATGTGGCTTTAGCCAACGTCATCGGTCCGGTACGTTCTTACATTGAAGCT GGAACCTTGCTGAATCAAGAAGGCACTTCGTTGAATCTGGTGCGTAGCTTCCCCGGCAAAGGCATTAAAATC TGGGGCTGCCGCACTCTGGATAACATACCTCATTCTCCCTGGCGTTATATCCAAATTCGCCGTTTGGTTTCCT ATATCGAAGCTCATATAACCCAACTTGGCCGCGCCTTTGTCTTTGAACCCAACAACGCCATCACCTGGATGAA ATTTAAAGGTCAGGCCCACAACTGGCTACGTCAATTATGGCTAAAAGGTGGATTACGGGGCACTCAGGAAGA TCAAGCATTTGAGGTGTTACTGGGTGTTAATGAATCCATGAGTGAAACGGATATCTTGGCCGGAAAAATGATC ATGAAAATCAGGCTGGCGCTGTTAATTCCGGCAGAATTTATTGAGCTGAGTCTGACGTTTGATATCCGTAACA ATACCGTACCTAGCTAATCTAAACAGGGGAAAAACATGTACAACTTATACACCCCGTCAGTATCTCACCGTTT TATCGCCAGTTTTCTGTTTAACAACATTCCCAGCCCACTTGATATCGCCTTTCAGCGTATATCTGGCCTGAGC CGAGAACTGCAAACCACCCAACATAGCCAAGGTGGAGAAAACGCCAGAAACGTCTGGTTATCCGAGAAGAT CCAACATGGCAGCCTGGTGCTGGAGCGCGGTGTTATGACCATCACTCCCCTCACCTTGGTTTTTGATCGCGT GCTGCGCGGTGAAAAAGCCGTGTATGCCGATGTTGTCATCATGCTACTGAATGAAAATGCGTTACCCGTGGC GAGCTGGACAGTCAGTAACGCGCTACCGGTTCGTTGGTCCACCAGCGACTTTGATGCTAATAGCAACACCGT ACTGGTGAGTTCTCTGGAATTACGTTATCAGGATATGCGCTGGTTAGGAGTAAAAGCATGACGGTAGAAATTA AAGAACTGATTATTCAGGCTAAAGTCACCGATTCTACGAGTGATCAACTCGCCCCAAGAACATTAGCCCAAGA AAAGCTGGATAACGCCCGTTTGATTGACATAGTGAAACGGGAAGTGTTAGAGGCATTACGTGAAGGAGGCCA TCATGAGTTTAATTGAACGTGGTTTATCCAGACTCACCCTAACCGCTTTTAAAGACCGAGAAGGTAAAGTTTC CGTGGGTCGCTTACAAGCCATGTATAACCCCGATACGATCCAGCTTGACTACCAAACCCGCTACCAACAGGA TGAAAGTGTTAATCGTGCCAGCCAAAGCAGCCGTTATGTATTATCCCAACCCGCCGGATTATCCTTAGTTCTG CTGTTTGATGCCTCGATGCCCGATAATAACATGCCGATAGAAACCCAGCTTGCGACCCTGAAATCCCTGTGT GCGATTGATGCCAGCACCAAAGTACCCCACTTCCTTAAAATCAAATGGGGCAAAATGCGCTGGGAAAACAAA GGTTATTTCGCCTGCCGAGCCAGTAGCCTGGCCGTCAACTATACCCTGTTTGACCGGGATGCCACACCATTG CGGGCCAGCGCCACTCTATCTCTGGTAGCGGACGAAAGCTTTATTATTCAAGCTACCGAACGGCAGTTAAAA TCACCGCCGGCCACTGCGGTTAGCGTAACTGATATGCTCTCCCTGCCTTTGATTGCTTTAGATGCTGGAGCG TCTCTGGCTGGTGGCATTGATTATCTCTCGCTGGCCTGGCAAAACGGTCTGGATAATCTTGATGACTTTACCC CCGGACAAACACTGCAAGCGCGGGGGGATGCATGAAGATACCCATGATAACCCTCAAAATAGGTGGCAAAA CGCTCAATCAATTGACTGTCATCAGTCTGACAATAAACCATCAAATCAATGGCATTCCCTCGACCAACATCAC CTTGGGGATCGCTGGCGATGCGAGCCATATTTTCGACACCAAAGCCCAAGCTGAACTGGCAAGTTGTCGCC CCAATAATGAACTCACCCTACAGATCCAAAAAACCGTGGTGTTTAAAGGGAGCATCGTTCGACAAGCACTTGA ACTGAAAGGTCAAGACAGCATCATTACCCTGACAGCAAAACATCCACTACAAAAGTTAACTCATAGCCTCCAT TCACAATTATTCAGTCAACAGAGTGATGAAGCGATTATCAGGAAATTATTCAATCAGGCGGGTATCCAAACAA CGATAAAGCAGGCTCCTCAACTTAAAACCGTTCATGAACAAATGGTGCAATTTCGTTGCAATGACTGGGCATT CCTAAAAAGCCGATTGATTGCCACTAATACCTGGCTGTTGCCCGGCAATGAATCGGTTACTTTGATAACACCT AAGGCCCTGAATCAATCGACAGTGCATACTCTTCATCGACAGGCCAGTGCTGAAGATATTGTGTTATTTGCAG CGGATCTCCAATGGAATAACCAATATAGCCCTAAAACGGTGAGTGTACGTGCCTGGGATATTGCTCAACAAA AGCTTTCCCCAGCAATTAATACCCAAAACAGTCAGCTTGGCAGTCATAAATTGGCCGTGGACAGTATCGCCG CACTGGCTGATAAAGAGTGGCAATGGGCTTACAGCTATCCATTAGATAATGAACAAGCCAAACACCTTGCTCA AGGCATTATGAATAACCTGCGAAGCCATAATATATCTGGCAGTTTTGAAATCGAAGGTAATCACCGTTATCAA CCGGGGGATGTCTTGGCGTTAAATGGTTTTGGTCAGGGGATGGACGGTCAAGGGATTATCACCGGAGTCAG TCAGATAATTAATCAGCGGCAAGGCTGGCACACCCTATTAACCTTAGGCATGTTACCCGATGTAGAACCGCC GGTGCCTCAGGTGAAAGAGTTGCATATCGGTATCGTGGAAAAATACCAGCAAGACCGCCAATCACTAAGCCG TATCCCAGTCAGAATACCCGCATTAAACTTGACCAAAGGTGTCCTTTTTGCCCGGCTAGGTAAACCTTATGCC AGTCATGAAAGCGGATTTGCTTTTATCCCGAACCGGGAGATGAAGTGATTATCGGATTCTTTGAATGTGATC CTCGTTTTCCAGTGATATTAGGTTCCATGCATAATCCGAAAAATAAACCACCGTTAGAACCCAGTGAAAAAAAT CCGGTGAAAACTTTAGTTATCAAGCAAGGGGATAAACAACAAGCATTAATATTCGATAATAAAGAAAACACGG TGGCACTTAATAGCGGCGAAAATAAAGTCTCTCTGCAACAGGATAAAAACATTACGCTCAATTCAACTAAAAA TCTCATCACTCAGGCCCAAGAAATTAATATACAAGCGGAAAAATCTCTGTCAGCCACAGGAAAATCTGGCGTC GATATTAAGGGCGCGAAAATTAACTTAACCCAGTAATGAGGTATTGAAATGACAAGCCAAATATTAGCCAATA TTTACGGTTGCGGCTGGAAATTTCCGCCACAGTTTTCTATTGAAACTGGCGTAGAAATGGCCGAAGGTGCCG AAAACGTTCGCCAAAGTATGAAAATCCTTTTTTTAACTGAACCCGGTGAACGAATTATGCGTGAAGATTATGG TTGTGGTCTGAATGATTACATGTTTGAAAATATCAGTGATGAATTATTATCGGAGATTCAAACCCGCATTGAAG AACGAGTATTGCGCTATGAACCCCGTGCTGAAATCACAGATATCCAAGTAACTCAGAAAACAGACTCACCGA ATACTTTACATATTCAAGTGACCTATGCCCTGAGAGGCAGCCAAATCAGTCAACAGCTTGAAGGGGTTCTTGA GATCAACGAAGGTCAGGCAAAGGTGAGTCTATGAGCAAACAACTCATTATTGATGGCGACAGCCTGCTATTC GAGCCATTATTCGGCAACCGGCAGGTCACTATTTTGATGCCAGCGACCATCAGAGGCAGCGGACACGCGCA AATCCAAGGCAGAAAGATAGCGATTGTCGGCGATGAAAAAAAGGTACAACTTCAAGCGCAATACATTACCCC AAGCCACCCGGTACCTGGCATAGGCACAGTTACCATTGCTCAATTAGATACCAGCCAGCAAGTCAACTTTTG CCACAGCCCTGCCACAGTGATAGTTGTCGGGCAGCAATTTACCGCTCGATTTACCCCATCACAGCCGGCAAT TAATCCGTCAACCGGGCCAGATGTCACAACACCCAGTATGGGCAAAGGCCGTTTTATTGCCAGTCAACATAC TATCAACGCCGGATAAATAACTCTGCAAAATCATTATTCAATAACGTTCCTATTCTGCAATAGCTATCAGCAAT ATATTCAAATAACAGGTGGTATAATATGGGACTCACCGAATTAAAAAATAAACTCTCTGCTATCGTACTCGATA CGGATTTTAAACTTGATGAAAGAAGTACACTGGATATTTTAAACTGGCTACAAGAATATGCTAAAAAAATCCCT TTCAATCAAGAGAAAAAACAGTTCTGGGATAGTTTCTATTTTATTCAGGAAAATAGTCCTGAGAAATTAGCCGA TCTTTACCAAAACGTTAATAAAACGAATGGCCATTTACCGGCCCATCAAGCTTTTGTTTTAGCCTTTTTAAAAC TTTTAGAAACCACCAAAGTATTATTTAATACTTTTCCGGCACGACATCGTGATCTTTATTACCGGGAATTATTA GGTCTAAAACCCAGAAATGCCCAAGCAGATAGTGTTGCTTTAGGCATTACCTTAAATACAGATAACACAGAAC ATCTTATTCCTAAAGGAACCTTGTTCGATGCCGGGCAGGACAGGGCCGGAAATCCGCTACAATACGCATCAA ATGCAGATTTACTGGCGAATCAAGGAAAATTGAGCGATCTGCGTTGGTGTCGAAAAGATAATGATAGCTGGC AATCTGCAATACTACTGAACCACTCAGATAATATTGAATTACCTGAAAACAGTATTCGACTTTTTAGTCCAACG CCGGATGATATTCCCGTTTTATCCGGTTATTTGATAACTTCGTCTTTATTTGCTATGCCAACGGGGGAACGCA GTATTACATTGACTTTAGCAGATAATTGGCATGGTGATATTAAGCACATCACCGCTAAAATCAGTTCGGGAGA TCACTGGCTTTCACTATCAGTAAAAAAAGAACAAGACAATAGTATTCACTATCTTAAACTTTATTTATCAACCAA TGATGACCCCATCGGTCCTCCTGATGCTTTGGATAATATAGCGTTTGATGTACCGGTATTAAAGCTGGGCACT GTTCAGGGACCTATACTACCCAAGATTACGGGTATTGAAATTAGCATTAACGGCAACAGTAATGTACATTATT CCTCTGATAACGGTATTGAAAAAATAGATGCAGCTAGTTTTCCCTTTGGACAATCACCGTCACCAGGTTCCGG TTTTAATCTGATTGCCCCTGAATGGTATGGTACAGAAAGCGCCAAAATTACTCTTACTCCTCAATGGACTGGA TTACCCAAAGAGGGGTTTAAAGAGTGGTATCAAGGATATAGTTCTACCCCCGAAAATAATGCATTTAAAGTAC AGGCTTATTTAATCACACCTCAAAAGAGAGAAAAATTTAATGAAGCTCAGTCATTATTTAATGAAAGTAAAGAC AAGAAACCACAAGGAAAAAGCCTAACTTTTACCTTACCTGCAATGGATTATTCCTTTGCAAACAGCCCATCAT CTAATAACTGGCCCGCATCAATACGCATAGAACTAACCGAACAGGATTTTATGCATGCCCAATATTGGCAAAA TCCTACGGGTAAAAAACAGCCCTATACCCCCAAAATGAACACATTACAAATTCAGTTCAGTGCCAAAGTTAAA CCCGAACAATTTTCCGTTTATTCTCTCACGCCTTTTGGTTGGGGAAAAACAGGAGAAAATAGAACATCATTAA CCCATGATACATTCTATTTAGGTTTTACCGATGTATTACCAGGACAAACTTTATCCCTGTACTGGCAGTTAGAA GGTATTAAAAAGCTCCCTTTATCCTGGTCTTATCTGAATCAAGAAAATACCTGGAGTCCATTGGATAATCAGGT GCATGACCAAACCCACAACCTATTTGATCGAGGAATCTGGCGTACCTCATTGCCACATGATGCTTCAAACCAA GCCTCTCAAATGCCAAAAGGACAATATTGGGTGAAGGCACACATTTTACAAACGAATCAAGCAACCCTGACT GATCTGTATTGGTATCGAAAAGATAATGATGTCTGGAAATCCGCAACACCTCTTAGCCTTTCAAATAACATGAA ATTACCCGCAAACGGTATTCAGATTTTTAGCCCAACATCTCATGATGTTCCAGTTCGATACGGCTACCTAATTA CTTCATCTTTATTCTCATTCCTCAAGAAAGGACGCAATATCACATTAATTTTAGCAGGAGATAGCTGGGAGGG TAATCCTGAAAACATCACCGCTAAAATCAGTTCAGGAAATCACTGGTTAACACTATCCGTCGAATATCTGAGT AATACTAATAGTCTTAAGTTGCAATTATCAGATAATAATAATGATCCCATCAGCCCCCCTAATGCTCTGGATAA TATGACGTTTGACACGCCATTGTTAAAACTAGAAGCCACTCAGGATTTCACTTTGCCCTGGATTTATAAGGTAT GCGTTAATAGCAACAATATACTCTCTACCTCTGACAGCTCAGATGCAGCGATTACTCGTTTCCCCTTTGGCCA ATCACCATCGTTGGGTTCCAGCTTTAGTCCGAAAATCGTTTTCCCGGAATGGTTTGAATCTGAATACGCATCA GACACCACGATCACGATTACCCCTCAATGGGTTAACCTGCCCACAGAAAACTTTTCATCGTGGTATGACGGA TATATTAATAAACCTGCCGATAATAGCGTATTTAAAATAGAGGGTTATTTACTTACTCATTATCAGGGAAAAATC AAACTCACAGAAGCTGAGACAGGAAGCGAAACCCAAGCATTATTCAATGGAAACAATGCACCACAAGGAAAA AGCCTGACTTTCACTTTACCTAATAGGTATAACTTCTATCCGCGCAACCATCAGTCAATGAAGATAGAAATAAA ACTCGTTAAACAAGACTTTATGCACACTCAACATAAGAGCAATCCCACAGGCAAAAAACCACCCTATACCCCG CAAATCAGTGCCTTACAGGTGGAATTCAATGCTACAGCTTTCCATCGAAAATTCTCCGTTTATCCTCTCACGC CTTTGGCTGGGGCAAAACAGGAGAAAATAGCACACCATTAATTUATGATACATTTTATTTAGGGTTGAGGGA TATATGAGGAGAGGAAAGTTTCTCTGTATTGGGAGGTAAAGGGGGTTAAAGAGGTAGGTTTGTGTTGGTTTT ATCTAAGTGAAGAAAATAGCTGGAAATCATTAAATAGATCAACTTACAACCAAACCCACAACCTGTTTGAATCA GCAGAACAAAGTATCCTATTACCACGGGATGCTTCAAACCAAGCCTCTCAAATGCCATTAGGACGGTATTGG CTGAAAGCACAGATAGAACAGGAGAAAAAACAGATAAAGATAGCGCTTCCTGATTATTATCCAAGAATCAGG GGGCTGTTGTATAACGCTACCATCGCCACTTTAATCAACGCTGAAGCTGTTGAGCAATCTCACCTTATCAACG GATTGGCTGCTAACAACATTAAACAACCGGTTAACTCATCCGTTGCCATCAACGAAGTTATTCAACCCTGGAC ATCCTGGAACGGTCGCCCAAAAGAAACCGAGTCAGCATTCCTGGCACGAGTTCCTGCCCGGCTCTCTCATC GTAAGGGAGTGGTAAGGTGGGGTAAGATTGGGAGTTTATTAAAAGAGAATTTTAGTAGGTTATTGGATGTGAA ATACCCTTCTGTCAGTGAATTAACCAAAATTCCAGCGCCAGAAAAGCGACAATTAACCATCATCCCCGACAAC CGCTATAAAGATAATGATGATTCACTACGCCCAGTATTGAACCAAGCCAGACTGACCGAGATGGTCGAATGG TTAGATCGATTAAGTAGCCCTTGGACAACTATTGAAATTAAAAATCCCACATATGTTAACGTTCTGATCCACTA TGAACTGATATTTACCTCGGATGTTAACCCCGATTATGGCCTCCATCAGCTACAACAAGAACTCAGTCGAAAA TATATGCCGTGGGGAGAAAATGCAGCTATTGGCGTAACACCCGGTAATCGTATTGACTACTTCCAGTTATTAG CCTCAATTCAACAATCACCGCTGGTTGAACGGGTCACCAACTTAACGTTAAAAAAAGGCAGCCAGCCTACCG TAAGTGAAAGTATAGAAGGGGGGGATGATGAAGTAGTGATTTTAGTGTGGTGATAAAAAGTTGGGGAAGGTAA GGAATTAACAAATGAATAATCGAGATATGCTATTTCCTATCATTAAAGACGATATTACCTTTGATTCTTTATTCG CCCAGGCAAAAGCCGTTATTGAACAACAATCGGGGCAGCTCTGGAATAATACAGGTGAAAATGATCCCGGCA TTACTTTATTAGAAGCCTGTTGTTATGGCGCATCCGATCTGGCCTATCGCCACACATTGCCACTGCGAGATTT GGTTAGTGGTGAAGAAAATGAAGGAATAGATGATGGGATTTTTGGGAAAGAATTGGTGGAGAAGAAATAGTG ACCTGCGGCCCAATTACCGCGGAAGATTACCGTCGAGCTTTGTTAGATTTGCGTAGTGATAACACCGTTGAA GGTTATTTTTTGTTTAATGATGGAGAGGTGATTGGTGAAGGGGAAAATGAAGGGTATTGATATTGGTATAAGAA AGAAAAAGGGGAATAGAGTTTTAGTGAAGAGGAATAGAGGGAAGAATTAGAGTTAAGAGTGAGAGGAAAGTAT TGGGTGTATTTAGTTGGGAGTGGGAAAAGGGAGGTGGATAAGAGGGTGGGTGAAGAAAGAGTGAAGATTTTTG TGAAAGATAACCGAAACTTAGGAGAATCGGTCAGTAAAATTATTTGGCTAGAACCCATTAAACTGTCATTGAA AATTGATATTCAGCTTGATGATGACGCCAAAGATATTGCTGATATATTTGCTAAAGTTTATATGATTGCAGAAC AAATGGTGCTTGAAAAACCATTACGTTATACCACTCAAGCGATGAAAGAACTGGGTTACAGTCAGGAACAAAT ATTTGAAGGCCCTTATTTACACCACGGTTGGATACCGAAATTACCTCAAACCAAAGATTATACTCACCCTACC GTATTAAATCTCAGTCCTTTAATTAATCAGTTACTGGCTATCAAAGGGGTGAAACATATTACCCAATTTACATT GGATAAGCCTGATAAAAAAATTTCTAAGTTACCAAATGATAATTGGTCTTGGGAAATCGCTCCGGGATATTAC CCAAAACTATGGGGAGATACTCCATTAGAATTAATTACCTCACCAACAAGCCCACTCACCATCACGGCAAAAG GGGGAATTAAAATTGCTATTACTAAACAACAGATAGAAAAAAACATAATGACAGAACCACTAATTAATACACAG CCAGAATTATTGAACTGGGGTAAACATCGCAAAGTCCTGGATTACTATCCGATAAGCAATAAATTACCCGCTT GGTATGGATTAGAAAGTAATAGGGAAGAAGAGGTAGAGTTGGATGAATTTATGGTGGGTTTTGAAGAAATGGTA GGGAATAAGTGGGGTGAAGTTGGTTTATTGGGAAGAGTATTAGGTTTTAAAGAAGGAGGAAATAGGGTAGATG GGATTGAATGGGGTTTTAAAGAAAATAGGGTTGGTGAAGATGTTGATAAGGAGATAGTATGTAATTTAAAGAAT AATGCTACGAAAATCGATAATAATGCCGATGACTACGACAAGGAACTCGTTATTCTAGATTATTTGTTAAGATA TTTTGGGGGTGAATGTGGAATGGGAGGAGTATGAGGAGAGGGAGGAGAATGATGATTAAGAGAAGGTGAGAGT AAAAAAGATTTTGTATGTAGTGAGGGGGAATATGTGGGTGAAGAGGGAAAAGTGAGTTATGAGGGTAAGAATAT TCGGATTGATAAAGTATCAGCACTGCAAAAACGTATCGCTGCCCGATTAGGTCTGGGAGGAGAATGTTTCAA AGCAGAGCCTGACTTAGCTCACCTTCCTTTCTACCTCATTGAACATCGTAGGCTCTTACCAGTAAAACCTGAT ATAAAATTCTATATTGAGCAACAACCTAATTCTCTGGAAATTGAAAATGATAAATTAAAAATCACACAGAAAGAT TCAGCGGGTCGGTTACTGCAAGGTCAAGTTATTAACCTGGAATTTCGTGAGGGCTATGATGAATTTACATTGC TAAACTTAATGATAACTGAAGTGACAAGAGATACATTCACCATTAGCATTAATAATAGCCGTGATCTCAGAGAC AATCTGGACAAAGTGCAACACGCGTTTGAACAAACGAATAATCTGAGCTGGCACAATAGCTTAATATGGATGG AAGATATGGATTATCAATTGGTTTATGCCAATGGAGAACAACTGGAAAAAGCGGAAAATGAACGATGGATTAC CATTAACAATCAAAGTGCTTTCCCTGCTATGATCGGAGAGAATGATGAAATCACACTAAAAATTCAATCCGATT ATGAACTTAAAACCAAAGTCGTGCGGCTTGATTATAACAACAAAAAAATTCTGATTATAAAAGATGCGACATCA ATAAATAATTTTGGGGGAAAAAGAGAAGGATGATATTATTGTTGGTGTTGTGTAAAAGAGAATGGGTAGGGATA TTCGGATGAATATAAATATGAACTTACTTATATTGATACAGATTCTACAAAAGAAAATGAGTGCTGGATTACTAT GAGGGATGGAAATAATTTGTTTTGTGGTGATATGATGGGAGAGAATGAGGAAATTATATTGAAAGGTAAGGGTA ATTATGAGTTTAAAACGCACGTAGTAAAATTTGATCGTATTAATAGACAAATATTACTTAGGAAAAATACAGAC GTGGAAAATAATTTTGGATGAGAAAAGAAGAGATGGGAGTATGGGTGGGATTTGTGTGGTGAAAAATATGGGG AAAGTGAGGATTTTTGATTTGTTGTGAGTGGAGTAGTGAATGGAGAATTAATTGAGAGGGGGAGAGTGGATGT CTATAAATTAGAGTCTTGGGTAAAAACTGAGATTTTATCTGAATTACCCGCGCATATCTCACTCGTTATTCATT GGCTATCATCGGAAGAATTCGAAAAATTTGCCAGTACTTATAAAGTTTGGCAAAATAATGGCGCTCCTTTAGG TGATCACGCATATAAAATTCTAGAAACATTAACACTTGGGAAAAAACCTTCTACTTCAGCAAGAAGGTCCAGC AGCTATATAGAAGCACAGTAATAATTCTTACAGAACATTAACCCATATTTATCTTATAATATCAAACATCATAAA AACAATCTTCAGCTCATTATAATGACATATTTCATACTCAGGTTTCTTCATATCTGTTAATTACAAAGAGAATAT TAATATGATCTCAGCACCAAATCTGTTAAATCGGATTATCATTACTATTGAAGCGAATAACGCACAGGCAGCTA AAAAAGTATTGCATGGCTCCCTGCTTAATCAATCCAGTATAAACAAACTCTTTGATTCATACTTTAACCAATAT GTTGTAACAGACTATCTACCTGAAGACACTCACCCTGAATCTTGGCGAAATACGATTAAATAGTTTTAATTC ACAGTTTGTTATTCGGCTTAATACTATTCTGAGTCAAGCATTGAGCCAATATCAGGTAAATAATCAAACTGATA TTGAGAAATTTATTTATTACTTATATCGAAAAGATTCTATATTAAACCCAATAGAGGAAATCAATAATCGTGAAA TTACTGACATCAATATTAAGCAATTAATTAACCAATTACCCCAGATACAAAACAATTGGACACTATTATTGGCA AAAAGCTGTTTATCCACACATAGCCTGAAAAAACTCCTGGCTATCAAAAAAACAGCTTTATTAACCGCCATTAA TCGTAAATTATCTGAAAAGATCAATATATCACCCTATCAGCAGGAATCGGTTTCCACCTGGCAATTGATACTGA ATGCGCTGAAATATATACAGCGACATAATACACAGGAAATACCTGAACCCGATGCGAAAGTCATATCACTCAT TACAACGGAACTCAATGACAATGCCATTAATACAGCACCAATTATTGCATTATTTCGCCAAGTTATAACCAACC ATTCCCCACTGAATAAGTGGCTGGAACAACTGTGGCAAACAAAGCGAATTTCACAGTTATGTAAAAAACAGCT GTCAATTGAAGAATACCAACATCTATCGGAGCGCTTTATTGCCAAACACGGGAATAAAAATAAATCTGATAAA AAATCATCCATGACTTCCGAACCGCTGTTATTACCTGAACACCCTCCACCACGTCAGGTCAATAATGCTGGAA TATTAGTTCTGTGGCCGATGTTACCTACTCTATTTAACCAATTCGGCCTGTTTGAAAAACAAAAATTTATTCATC GTCAAGCTCAATTTAGGGCTGTTAATCTACTTGATTATCTCATTTGGGGAAACGAAGAAACACAGACAGAACG AAAAATATTGAATTGCGTTCTGTGTGGGTTAATTGCCGATGAGGACACGGAATCAATCCCTATTGAGCCAGAA AAACAACAGGTAATAGAACAATGGTTAGATGCAGTTATCAGTCAACTTCCTGCCTGGAAAAAATTAAGCCGCA ATGATAGCCGCCAATTGTTTTTACAACGCCCGGGGGAATTGCTGACAAATGAGCAGGAAATCAAAATTACGG TACAACCTCAACCATTCGATGCACTGTTAAATAACTGGCCCTGGCCGTTAAATATCGCCAAACTTCCCTGGCT GGATCGCCCTTTATTAATCAACTGGTAAAACATTGACAAGGTTTATATGAAAGAACATCAATATAGAATAGTCG ATCTACGCTGGATTTATTCCCATTTGGAGCGCATCGATCTGCTGTTACAACGTCACTATTACCAAAAGAGAGA CAAATACGATTCATTGCCAGAAAGTTTTTGCTTGAAGAAGATGAATTAGAACAACGTCTAGCAAAACCGTTG GGTATTCCTCATTGGCTAACAGCAAATACCGGCGCTGGTGATACAGAAACAGAAAATCATTCTGCTTCCGGC ACATTATCACTGCTAGTCACGCGTTTTAAACTCACTGAATTTGAACGTGATGTGTTATTGCTAGGTTTATTACC GCATTTTGACAACCGCTATCATGCGTTATTTGCTACTCTGCACGGTAACAGTAAAAAACAGTGGCCCAGTTTT GATTTAGCGATTGAATTATTTAGCCAACATCAAAGTAACTGGCAATTATTTCAACACCACTTTTTACCGCAAGC TCCATTAATCAATCACCATTTATTACGACTCAATAACCAAGAGGAACCCATTTGGCTACAAACTCAATTTTTAA CTCACAATGCAGTCTGGTCTTTTTTATCCGGTCAGCGCGTCATTTTACCTCCCTTAATATCCTGCGCTTACTG GCATATTCCAACCTCACAGACTTGGTATCCACCAATCCTTGGTCATGCATTTGAAAAAATATTGCTGAATGAAA CGGACGAAATACGCCCGCTGGTGGTTCTTAAAGGAAAACAGGACAGCGCCAGAGAACTGGCAGTCAGTAAT ATTATGGGAATTCACGGCATTAACACTTTAACGTTCGATTTATTTCACCTGCCAGATGAAGAGTGCACCACCT CAATACTCAATCTGCTAATAGATGCAATACGAGAAACCCGGCTACATAATGCCTGTTTATTAATCCGTAACTTT TCTTTGCTGGCAGAGGAAAAGAGAATATCGCATAGAGAATTATCAGCTCTACTGAATCAACCCAAATTACGTG TGGTTTGTCTGGCAGAGTCAGAAGAATCATTAGCATGGGTTAAACACCTGCCGATAGTGCAAATTAATATGCC ACCGGCGACGCTGGCAGATAAAAAAACGATGCTGGAAGCCAGTTTGCCAGATAATGTCACTAAAGGAATTAA TATAACTCAATTATGTCAACGTTTTTCATTTACAGCAGAAACATTACCGTTAATTATCAAGGAAGCTCATCAATA CCAAATCCTCCGACAACCGGAAGATCAATTGAAAGAATCTGATCTACGTAAGGCATTAAATTGCCGCGCCCA ACAAAATTTCGGTAAATTAGCCCAGCGTATGACACCAAAACGAAGTTTTAATGATTTGGTTATTTCCGCTGACT TAACTCAACAGTTGAAAGAAATCATCGCAGCAATTAATTACCGTGACCAAATTCTGGGCGCAGGTTTTCGGGA AAAAATCAGCTATGGTACTGGTATTAGCGCCCTATTTTACGGTGAATCCGGGACGGGGAAAACCATGGCCGC AGAAGTGATTGCCAGCTATCTTGGTGTTGATCTGATTAAGGTAGATCTTTCTACCGTGGTGAATAAATACATC GGTGAAACCGAAAAAAATATCTCCCGTATTTTCGATCTGGCCGAAGCGGATTCCGGGGTGCTGTTTTTCGAT GAAGCCGATGCCTTATTCGGTAAACGCAGTGAAACCAAAGATGCCCAAGATAGACATGCCAATATTGAAGTTT CTTATTTATTACAGCGACTAGAAAATTATCCGGGATTAGTGATTTTAGCGACTAACAATCGCAACCATTTGGAT AGTGCGTTTAATCGCCGCTTTACCTTTATTACCCGCTTTACTTATCCCGATGAAGCATTACGCAAAGCAATGT GGCAGGCAATTTGGCCTGAACAACTTAAGTTATCAGATCAACTTGATTTTGAGCATTTGGCTAAACAGGCAAA TCTGACCGGTGCTAATATCAGAAATATTGCCTTATTATCATCAATATTAGCTACAGATAATAATAGTGATCAAAT TGAAAATAAACATATAGCGCGAGCATTGATACTTGAATTAAATAAAACGGGCCGATTGATTTTTTAATCATTTA TACCCAATAAATTTCGAGTTGCAGCGCGGCGGCAAGTGAACGAATCCCCAGGAGCATAGATAACTATGTGAC TGGGGTGAGTGAAAGCAGCCAACAAAGCAGCAACTTGAAGGATGAAGGGTATATAGAATTGGAGTGAATATG ACAAATATAATTAACCCTAATAATGCGATTCTTGAAGTTAATAACGCATTAAATGATATTTTATCTCAGTATTTA ACTAATATTGATATCCGCTTTGATCTACCAGAAATAAATTCAATCCCATCAACCCCTACAGTGAGTATATTTCTT TATGATATACATGAAGACCTACAATTACGTTCTGCTGAACCAAGAAGTTATCATCCTACCACCAGCTCATTATT GCCGGGATGGGTAAATATTAATTATAACTATTTAATTACTTACTGGCATTCAAGTAATCCATCAAGCGACAGTT CTACCCCTGATAGTCAACCCAATAATCAAGCGGCACAAGTCATGACTGCTATTTTAAATGCATTGGTTAACAA CCGACAATTACCTAAAATTCCTGGCGCATATACCAGAGTCATTCCACCTCAAGAAAATCTAAATAGCTTAGGT AACTTTTGGCAAGCGCTTGGCAATCGCCCTCGCCTTTCTTTATTATATTCAATTACCGCACCGGTAAAACTGC AAAATATTAAAGATGTCATAAAGCCCATTAGCCAAATTTCCACTTCTGTGGATCAAAAATCAAATCTGGATAAT TCGCAAATCAACCAAGCCTTATTTAGCAAATTGGGTGCCGATTTAGGTGGCACACAAGATGTTCGTCTTGCTC TTGCGAAAGTGAATCTGACAACCAAACCTGCTAAAGAAAATAATGAAAATCAAAATAATAAAAATGTAATTATT GAAGTTTCTGGCATTACCCATTTGGATTATTTACCCAGAATAAAAGGTATTCTTTCAACATGGGTAAATAGTCA TAGTGCTGTTGTTAGGATAAATGATATTGGTATTATTGTTTCAGAATATAAATATGATAAATTAACAGGCGTTTA A (Photorhabdus asymbiotica strain ATCC43949 PVCPaTox operon, pvc1 - pvc16} SEQ ID NO: 95 ATGAATACAGCTCAAGAAATTATTAACCGTTTATCGGGGAGAGCCGTTACGCTTGGTTGGGATGTTGTTATTG CTTATGACCGAAAAAAAATTAACACTCTGTTAGAGCAACAATATGTTGAAAAGGTAAAAAACGGGGAGAACTT CCCGCTTATCAACTGGGAGAACCAGAGAAAAACACTTCAATTTAAAGATCTTCAATTAGGTGTTCCACTTATTT CTTTTGAGAATTCAACACTGGAAAATTCAAGGGCGCTTGCCACGATAGAATTTATTTCAGGAGCTATTATTGAA TTTAGTGACTCCGGGCAAATAATCAACTATAAGAAGATTGAACCTAGTCATGGTTATGGCATGGTGCTGACTA TCGATCTCATGGCTGGTACAGGTTCAGTAGAAGAACAAGGTCGGGTGATAATAAATCTTAACGAAGGCGCCA TACTCGATTTGCATGTTATCCAACAACCGCCAGCAGAAGTGGTAGAATTTTTCCGCACTTGGTTGATGGCTAA TAAAATGACTTATGAATTAGGTAAGCTGGATCTGAGTAGTCAAGCTGGTCTAGTGCCTCGTTCTTTTCGTATTC GTACTCAGCGGGCGCCTGAAAAAATTCGTAAAGCGACGAGCGATGAAGGAAATGGCGCTGTTTTGTTGTTTG TTGCCACTAACTATAACCCTACAAGTGGAACTTTACCTGCCAAGGATTATCCGTGGCTAATCCCTGAGGAATA TTCAGGCGCATTGCTTATCGGTAATAAATGCTTATTTAAAGACATTCTGAAACCGAATCTGGATCAGTTGTTTG ATAAAGGGGAATGGACATTAAAAGTTCAGCAAACGGATTCTGATCAACTGCTGCATTATCTGGAGGCAAACTC TGCATATATAACAGATAAGCCTTATATGGCAGACTTTGAAGGAACTCAGGATGGAGTCTGGACAGGACGTTAT AAATTTGAGACTGGCCGGGGACATTATGGGGTGTATGAAAATGTACGCTTTCCTATCAATGGAATGTTGATGA AACCGGCTAAAACTGGATTACAGTTATCAATAGATTCACCACAAAGCCATCAATTTAATGTTGATTTCGGAATG AAGTGGTTCCATTGTGCTAATATAATGTGTGGTTATTCCTGGTTTAACGAGACTTACCCATTTTATCTTGATGG AAAATCATTTTATCAAGTTCATATTGACCCTGATAAAGAGGTGATTTATTTTACTGGGCCAGATGAAGATATTA ATATTGTAGGAAATTACAGCCCGCCTGCGTGGTGGCAATCTAAATGGCAAAAACATATCAGTGATGATTTTAC GGATATTTCCTCGGAAAAATTTAAGCGACTCAGTCAAATAAAATTGCCAGAAATATGCATGTTTGCCGTGAAC CATTTATTATTTCCTGGTCATAATACTTTGCTGTTGAAAGACGTTTATTTACCGGGTGATATGGTGATTTTCGG TGATATTAACCCATCACTTACCGCTTTTCGGGTTACGCCATTAAAAGCAACAGTGGTGGCAAAGGGAACCCAA CAATTTAAAGCCATAGAAACTAATTGATGATTATACCCTTCATCCTTCAAGTTGCTGCTTTGTTGGCTACGTTC ACTCACCCCAGTCACATAGTTAGCTATGCTCCCGGGGATTCGCTCCCTGGCCGTCGCGATGCATCTTGAAAT CCATAGGGTATATATTTAATTGGATAAGTCTTTTTTATTTTAACATTATAACCTGATTCTTTTTGGATAAAATTAA AGGATTATTAACATGTCTATTACACAAGAACAAATCGCTGCTGAATATCCTATTCCTAGTTACCGTTTTATGGT TTCTATAGGAGATGTGCAAGTCCCTTTTAATAGTGTTTCGGGATTAGATAGGAAATATGAGGTTATTGAATATA AAGATGGCATTGGTAATTATTATAAAATGCCAGGACAAATACAGAGGGTTGATATTACACTTCGGAAAGGCAT ATTCTCTGGGAAAAATGATTTATTTAATTGGATTAATTCCATTGAACTCAATCGGGTAGAAAAAAAGGATATTA CAATTAGTTTAACTAATGATACTGGCAGTAAAGTCTTAATGAGTTGGGTTGTTTCGAACGCCTTTCCGAGCTC ACTGACGGCCCCTTCATTTGATGCTTCAAGTAATGAAATTGCAGTACAAGAAATTTCATTAGTTGCTGATCGG GTAACAATTCAGGTTCCCTGATAACTAAAAACTTTAAGGAAAAATAATGTCTGTACAAACAACTTATCCCGGAA TTTATATTGAAGAAGATGCATCATTGTCTCTATCTATCAATAATAGTCCAACAGCAATCCCTGTTTTTATCGGTA AATTTTACAACTTGGATGGTTCCTTACCTAAAGTGGGAACATGTTCTAGAATTACCAGTTGGTTAGATTTCACT AAAAAATTTTCGGTAGCTCCTCCTCAAACCATTTCATTGATCGCGTCGCCAATTGCTGACACACAAGAAAGTG TACCCAAAGCAGTTCAATATACTTATAAGGCCGAGTTTGAAACCTCAGAAAATCTGGCAAATGGTGCCTATGC GGTACAACATTATTTCCAGAATGGCGGTGGTATTTGCTATATCATACCTTTAGTTAGCGTGAAAAAAGAGGAT GCTGCGATTGAGTTAACAAAATTACCTGAATTAATTGAAAGACAACAAGAGATTACGTTAATCGTCTGCCCGG AGGACGATAAGACGCTCACTGTTGATAGCAGTAAAAAATCGGATGTTTATAACAGCATCAATACATTATTGAG TAATAAGGTAGGTTATTTTCTCATTGCAGATTCAGATGATGGCAAAGCAGTTCCTGATACGTTGCCGGAAAAA ACTGCGGTCTATTATCCTGGTTTACTAACTTCTTTTACACAACGCTATGCCCGACCTGCCGATTCTGCTATCAA AGTGACCGGTATTACAAATATATCAACTCTGGCTGATATTCACACCAACTTGGCCGATGACTACTCAACAGCA AGTCAGGTTATTAATGATGTTTTGGAAAAAAATAATAAGCTCGCATCGTCTCCCATTATTTTACCTCCCAGCGC CGCTGTTGCTGGTGCTTATGCCGCTGTTGATGTGAGTCGTGGTGTTTGGAAAGCACCTGCGAATGTGATGTT AAGTAATGCCACGCCAATCATTAGTATTTCCGATGCGGAACAAGGTGTGATGAACCCATTAGGTATTAATGCT ATTCGTAGTTTTACTGGTAGAGGTACTTTGATTTGGGGAGCTCGTACTCTGGATAAAACGGATAACTGGCGCT ATGTTCCTGTACGTCGTTTATTCAATAGCGCAGAGCGAGATATTAAGTTAGCAATGCGTTTTGCAGTTTTTGA GCCTAACTCCCAACCAATTTGGGAAAAGGTCAAGGCTGCTATCAATAGCTATTTGCAGTCACTTTGGCAGCAA GGTGCACTGCAAGGCAATAAACCCGATGAAGCCTGGTTTGTACAAATTGGTAAAGGCGTGACCATGACAGAT GATGATATTAAGAATGGGAGAATGATTATCAAAATCGGCATGGCGGCAGTACGTCCGGCAGAATTCATTATTT TACAGTTTACGCAGAATATCGCCCAGTAACTTAGGTCTATACCCTATAGATTTCAAGATGCATCGCGGCGGCA AGGGAGCGAATCCCCGGGAGCATATACCCAATAGATTTCAAGTTGCAGTGCGGCGGCAAGTGAACGCATCC CCAGGAGCATAGATAACTATGTGACTGGGGTAAGTGAACGCAGCCAACAAAGCAGCAGCTTGAAAGATGAA GGGTATAGATAACGATGTGACCGGGGTGAGTGAGTGCAGCCAACAAAGAGGCAACTTGAAAGATAACGGGT ATATTTAATATGGGCGATTTATTGCCCATTTTTGTGAAAGGAAATGAGTTATGTCGCCAACGCTACCCGGTGT AACGATGACTCAGGCGCAGATAACAGCGTTCGGTGTCAGTACATTAAATATGCCCGTATTCATAGGGTATTGT ACGAGATTGCCTGCCTTTTCAGCGCCTGTAAAAGTAAACAGTTTAGCTGAAACAGAACAAATAATAGGGAAAG AAGGGCGTTTGTATGCTCTATTGCGCCACTTTTTCGATAACGATGGGATACAAGCTTTTATTCTGTCGTTAGG CGCACCTGCTGGGGAAAATGCTAATAGTTGGCTTGAGGCATTACAACAGCCCGATTTGTATGCGGCTGTTGC AGCAGAGCCGCTAATTACACTTTTAGCCGTCGTTGAGGCAAGTGAACTGAACCAAAAAGAAGGTAATGAGGC TGTGGAAGCTTGGCGACAGTACTGGAAAGCAGTATTAGCGTTATGTCAGGCACGCAGTGACTTGTTTGCCAT ATTGGAGGCACCAGATGATACCGCATTAATCAAGCGTAGTTTGCAGGATTTTCATCATAAGGCACGTCAGTTT GGCGCTCTCTACTGGCCAAGGCTAGAAACATCTTATCAATCCTCTCAGTTAAAAATTTTGTCTCCTATTGGTG CAGTAGCAGCGGTTATTCAAAGTAATGATGTCCGGCGAGGGGTAGGACATGCACCTGCCAATATAGCGTTAA AACAGACGATTCGCCCGATAAAGTCCCGCCTGGAATTAGAAGAGTTGTATGAAGAATCGGATGGTTCACTGA ATCTGATTTGTAGTTTTCCAGCTCGTGGTACTCGTATTTGGGGATGTCGTACGTTGGCGGGTATTGATTCACC TTGGCGTTATATTCAAACCCGATTATTGACTTCACACGTGGAAAGGCAACTCAGCCAGTTAGGGTGCATGTTG ATGTTTGAACCTAATAACGCAGTCACTTGGATGAAGTTTAAAGGCCATGCTGGGAATCTATTAAGGCAGCTTT GGTTACAAGGGGTGCTGTATGGGCAGCGTGAAGATGAAGCCTTTTCCGTTGAAATAGATGAAAACGAAACGA TGACTCGCCAGGATATTGATGAAGGCAGAATGATTGCTCGTATTCATTTGGCATTGTTAGCACCGGCAGAGTT TATCGCTGTGAGTTTGAATTTTGATAGTGGGTGAGGGATTGGGAGGAGTAGATAATAAATGGGAATATGTGGAT GAGAGTAGGAGGAGAGGTTTATAGGGGAGGGGTTTGAGATGGTTTTATTGTTAATTTTGTTTTTAAAGGTTTAG TTCCTTCTCCCGTAGATATTCGATTTCAACGTGTTTCTGGTTTAGGGCGTGAGTTACAGGTTGAACAGCGCCA TCAGGGGGGAGAAAACGCACGGAATCATTGGTTGGCTGAACGTATACAGCATAATAGCTTGATATTAGAAAG AGGGGTTATGGTCGTTACCCCTTTAACACTGATGTTTGATCAGGTGATGCGGGGGGAAACTCTCAATTGGGC AGATGTGGTAATTATTCTTCTCGATCAGGCTCAACGTCCGATAACAAGTTGGACCTTGAGTCATGCGCTACCG GTTCGCTGGCAAACAGGAGATTTAGATGCCAACAGTAACCAAGTGCTGATTAACACCTTAGAGCTGCGTTAT GAAGATATGCGCATTATAGGGGTAAAATTATGACTATCGAAATCCGTGAACTCATTGTTCAAGCCCGTGTTGT CGGGACTGATACCAAAACAACACGAACCGTTCCTTTATCTATTGTGCAAATGGAAACACTTATAGAACAACGT CTGGTTGAAAAAGTGAAGCGGGAGATATTAGACGTACTCCGGGAAGAACAAGGTGGTGGGTTATGAGCTTG CTTGAACGAGGTCTGGCTAAACTCACGATTACGGGTTGGAAGGAGCGTGAGCGTAAACATCAGATTGGTAAA CTAGAAGCAATGTATAACCCGGAAACACTTCAACTGGATTATCAAACTGATTATCTCCCTGATGTTAGCAATAA TGAGGTAAGAGTGAGTAAGGGGTAGGTTTTGTGAAAGGGGGGAGGGTTAAGAGTATGGTTGTTATTTGATGGG AATATGGCTGGTCTTACGACAACCGTCGAGTCCCAAATCACTACCCTCAAATCGCTTTGTTTAGTTAATGCAA GTAGTGATGAAGGGAATTTTTTGGAAATTAATTGGGGGGGAATGGGTTGGGAAAATAAAAATTATTTTGTTGGT CGGGCTAGTGGATTGTCTCTGACTTATTTGCGCTTTGATCGTAACGCAACACCATTGCGTGTGAGTGCGCAG CTCACATTAGTCGCAGATGAAAGCTTTGTGCTCCAGGATAACCAAGCCAAGTTAGATGCGCCGCCGGTATCA GTAGTTAATGTCCCGGATCTGACTTCATTACCTGCACTGGCGAATATCGCTAGCGTAACCACTATGTTGGGA GTGGATTATTTAATGTTAGCCCGCACCAATGATATGGATAATTTGGATGATATGCAGCCAGGTCAGACATTGC GAAGAGGGGAGGGATGATGAGTTTTTTAGATAAGAGTAAGTTGAAGGGATGAGATATGAAAGTGTTGGTTAAG ATTCAGGGAGTGGAGAAGGAACTCAACGAACTGATAGTAAGCGAATTGAAAATCTCCCGACGTATCAATGCC ATTCCGCAGGCAGTTGTAAAGCTAAGAGCGAAAGAGAGTGAAAGTGGTGTATATCAGTCTGATGTACAGCGG ATGTTGAAGAGTTGGGGTGGGGGAGTAAAGGGAGAGGTTGGTATTTTGAATAGGGGGGTATTGAGTGGGGAT ATTGTGCAGCAAAAAACAGAGTTAGTGTATGCGAAAACACACACTATCAAATTGGTGCTACGCCATGACTTAC AGGGGATGAGGGGTAATTTTGGTAGGAGAGTGTTTGGGAATAGGGGTGATGGTAAAGTGATAGGGGATGTATT GAATAGGGGAAGATTAAAGGGGGGATTTTGGGGGAGATGAGATTGGGATATAGATGATGAGGAAGTGGTTGA GTATGGTTGGAGTGATTGGGAATTTTTGTTGGAAGGGGTGTATGGTAGGAATAGGTGGTTGTTAGGTGAAGAA GATAAAGATAACACTCAGGGGAAAGTGACCATTATTGCTCCAAATTCTTTGCCCCTGAATGAGCGTTGGACAC TGCAACATCAGGCTGATCATCAGGCTATCCGGCTTTACAGCACGGAGCTGATGCTGGATAACCGGTTTGATA CAGCGGAGGCTGTTGTTAGTGCTTGGGATATTGATGATCAGGCATTACTCGTGGCGTGGAAAGAAACCCTTA GTGAAGTTGGGAAAGATGGGTTAGGGTGAGATAATTTTAGGGAGAGAAATAAAGATTGGAGTGAAGTGTTATT AAGTTGTGGGGTGTGTAGAAAAGAAGTTGAATTTTTAAGGGGTAGGGAATTAGTGATGGGGGGGTTGAGGGGG GTTGGTGGTTGAGTGAAGGTTGAAGGGAGTAGTAAGTAGGGTTTAGGGGATGAAGTGATGTTGTGAGGTTTT GGTGAAAATATGGATGGCTCACAAATACTGACGGGAGTGGATCATCGAATAACGGCAGAAGAAAGTTGGAAA ACAACCTTACATGTGGGATTAGAACTGCCGTTAAAGGCAGAGTATGTCACTCAGGTTAACGGTGTTCATATCG GGAAGGTTGGTGATTATGAATGAGATAGGAAAAAATGGGATGGTATTGGTGTTTTGATGGGTGGATTTGGAAG GAATATTGGGTTGTTTGGGGGATTGGGAAAAGGGTAGGGGAGGGAGGAAAGTGGATTTTGTTTGTATGGTGAA AGGGGTGATGAAGTGATTGTGAGTTTTTTGGAAGGGGAGGGTGGTTATGGTGTGATTATTGATTGGGTGGATA ATCCTAAACAACAGACTCCATTGCAAATCAGCAAAGAGAATAATCTCAAAATGTTGATGATTAAGCAGAGCGA TAAAGATGAGCAACAATTGTTATTTGATAGCCAGCAACAAACAGTCGCGTTAATCGGTAAGAAAAATATCGAG GTTAAAGGTGAGTATATCAACCTGACTAAATCAAAGGGGACTCGATAATGGCAAATACGCTTATTGGCCAGGT ATATGGTGAAGGATGGGGTTTTGGGATTAAATTTATTGGTGATAATAAAGAAAGGGGAGATGAAAGAGGGGGT ATTGTTATGGCTCAAGGGATTGAAGATGTCAGTCAATCGCTGGAAATATTATTTCTTACCGAGCCTGGCGAAC GAATTATGGGTGAAGATTTTGGTTGTGGTTTAGAAGATTTTGTTTTTGAAAATATTAGTGATAGGGTAATTTGTG CCATCAAAAATCGTATTCAGCAAGCAATATTACGTTATGAACCTCGCGCATATTTATTGAACGTTGATATTCAA ACCAAAGAAAACCAACCTGGACATCTGCTCATTCAGATTAATTGGAAATTACGTGGTAGTGATATATCTCAGC GTTTAGACGGAGTGCTTAGACTCCATTCAGGTCAAGCATTGGAACTGTTATGACCAATTATATTATTATCGAC GGGGATCTCATTCAAATAAATCCCAAATTTGAGGGTGATCGAACTCTTACGATTAATGGTATTCCTAAAATAAG CGGGAATGGAGATGCGCAAATTGAAGGAAAAAATATTTGTGTGTCAGGTGATCACTTAACTGTCTCAATTCCA GCCATTTATATAACCTCCAGACATCCTGTTGCAGGTAGTGGAAAAGTGAAAATTACAAATTTATCTGACGACC AAGTAGGAGAATTTTGTGTTAGTGGGGATGTTGTGATTATTGAAGGGAGTGAGTTTGAAGGTGAGTTTAGAGG GGATAAGCCGGCCACTAATCCAAGTAACCAAGATGCAGATAATCCTGCGCCTTCGAATGGGAGTGGGAGATT TATAGAGTGAGAGAAGTTGGTTAAGGGAGAAAAATAAAAAATTTTGGGGAAGGGGTTAATAAGTATGAATAAG GGGGGGGGATAAAAAGATGGATGTTGGTGAATTAAATAATAGGTTGATGAATGAGTTAGGAAGGAGGAATTTT AAGTTAGAAAGAAAGGAGGGATTAAGGGAATTAAAGTGGTTAGAAGGTTATAGAGAAAATATTGGTTTTTATGG GAATGATGATTATTTCTGGCATCAATTCTGGTTCTTAAAAAATCACACACCAGAAGCGCTCTTTGCTCGTTTGC AAGGTGAAAGGTTGGGTGATGGAGAATTGGGTGGTGATGAAGGGGTATTGGTGGGGTTTTTAGAAGAGGTTAA GAGGGGAGGAATGATGGTTGATAGTTTTTGAGGGGGTGATGGGGAATTGTAGTATGAGGAATTGGTAGGGATA ACGCAGAAAGATGCACAACCTGATCATGTGGCGCTTGGCGTGGTATTAAGTACTGGTATTGCAGAATATTTAT TACCGACAGGCACATTAGTGGATGGTGGACAAGACAGCAGCGGAAATTCACTGCAATATGCGTTGGATACCG ATTTATTGGTTAATCCAGGGCAATTAACAGATGTTCGCTACAGCTATTTGGATCATAAGACCTATAAAATCTTC ATCTTGCAAGATGATAAAGCGAATATCAGTTGGCCCTCTTCAGGCGCTCGTTTATTTGTAGCACCTGAGGGCA ACGGACAGGAAAAGGCACCTGAACAAAAGTTGGCACTTTACCTGGGATTTGATGATATACAGCCAGGGCAAA CTCTTCTTTATTTTGGCAATTCATGCATCAACTCCCCTGACATTAAAATGGTTTTATCTGAACGAGATAAATA ACTGGGTGAAGCTAGATAGTGTCAGAGATAACACGGATGGCTTTTTTATCAGTGGATTATGGCAAGCGATATT ACCTGATGATGCGGTGAAAATGTATTTTCCAGAGACAACTTCTGTAAAACGCTACTGGATTAAAGCTGAGGTG GAATCGCTTACTGAATCTGGCGATTTGTGGCAACCGCTATTAGAAGGCATCTTGTATAACGCTCAAACAGCAA CGCTGGTTGATGCAGACAACACAGATGAAAAGCACTTTCATGATGGGCTGATGCCTTTTAGCGTGCAGCATT TGGTCAACACCGTTTCAGAGGTAAAAAAAATTGAGCAGCCCTGGTCTTCTTGGGGGGGAACGCCACAGGAA GACACTACTGATTTCTTCCATCGAGCGGCAACACGTCTTCAGCATCGCCAGCGTGCGTTAACTTGGGATAAC CAAATTGCCATGTTGAAGGCTGAATTTCCGCGGATTTATGATGTCATCTCACCAAATATCACGTGGATGAACC AACTTCAGACATCAAATACGCAAACGCTGATCGTTATTCCTGATGTGAACTACAGCGACAACAAGGATCGCTT ACGGCCACAATTCAGCCCTGCCAGCTTGCGACAAATGAGTGACTGGTTACAGATTCACACTAGCGCATGGGC GAATCCACAAGTGGAAAATCCAATTTATATTGATGTCTCTGTGACCTATGAGGTGCAATTTAGTGCGGGTGTG AATCCTGATTATGCCCTCCGGCAATTACAACAATGGTTGAGTTCAATTTATATGCCATGGTATCACGCAGATA AAAAAGGTGTTGCCGCTGGCGATCAAATCGATTTTTACCAACTGTTTGCAGATATTCAGCGAGTACCTTACGT GGAGCATGTCAAAACATTGACATTGACCACAAAAGACACCTCATTAACCAATGGCGGGGTTATTAAGGCACA GCAAAATGAAGTGCTGGTGTTGGTATGGCAACAAGGAGAACAAATTAGGCAGGGAGAATCGAAATGAGGCA GCATAATGAGTTATTTCCTGTAGTAAAAGACGCGATAAGCTTTGAAAACCTGCAAGCTCAGGGTGAGAAGGTT ATTAGTGATCAGTCCGGTAACATATGGAGCGATAAAGATAAACATGATCCTGGTATAACATTACTAGACTCTTT AAGTTACGGTGTTTCGGATTTAGCGTATCGGCACTCATTACCTTTAACCGATTTATTAACCATTGCTGGAAAAG ATACGCTTTTTCCAGCCGAATTCGGGCCACAGCAGACGCTAACTTGTGGCCCTATAACACTGGATGATTACC GGCGTGCGTTACTTGATTTACATGGTAATGATGCATTTAAAATATCAGCTAGTGACCCCAGAGACTTTTTGTTT CAGGATATACAGTTAATTTGTGAGCCAAAAAGTAAGCGTTATAAATACTATTTCAATCCCGAAACGCTTGAATA TACATTCACGCCACCTTCAGGGGATAAATTTAAAACTTTAACACTACGAGGGAATTATTGGCTTTATTGGATAC CAACCCGTTGGGCAGGTAAATCAGCTAATTTGCCGTTAGTTAAGCGGGTGATGGAAGATTTTCTCCGTGAAA ATCGAAATTTGGGGGAAAATGTTGTTCAAGTGACACGGGTGATATCAACGCCTATTTATCCTGAGCTGGTCAT TGAGCTGGCGGATGATATTACAGATGCGGCATCAGTATTAGCATCAATCTATATGCTATTAGAACAGTGGGC GATGCCGATGCCTGCTCGCTTTACTACCGAAGCATTACAGGCCAAGGGATTAACAAACGAAGAGATCTTTGA TGGGCCGTGGTTGCGTCATGGTTGGATACCTCAGTTACCGACCTCTCAAAACTACCATACAGGCATGGTTCT GAAGATGAATCATCTGATTAACCAATTGCTGGCGGTTGAAGGTATAAAGCGCGTAGTTAGCCTGACGTTGCC AGAAACAGAATATTTGCATCAGATAAAAGATGATAATTGGTCCTGGCAATTAGATGTTGGTTATTATCCATTAT TATGGGGAGCTAATCCACTAGAGGTAATTACAGAGAAAAATAACAATTATGTCAAATTGTTCGCAAAAGGTGG GGTACGATTACAACCTGATCAGAAAAGTGTTGAGCGGTTATTATCACAGGAATCACTCATTAATAATGCTGCA TCCACGTTACCGGCTGGTAAGGTGCGTGATCTCAAAGCCTATACACCTATAAGCCGCAGGTTGCCTGCCTGT TATGGTTTGCAGAATACTTTGCAAAAGTTAAAACCTGAACAACGACACTTATATCAGTTCCTATTACCATTGGA GCAAATGCTTGCTGATGGATGTGCGCGGCTTGCATTTTTGCCACATTTGTTAGCATTTAGGGACCGAAGCGG AAATATCAGTGATACACTCTGGCCTTTCAAGAATACAGAGGACACAATTGCCCAACAGGTTCATCAGGAATAT GCCGGTACATTAAAAGCCTTTCAACAGCAGGAAATTAGCCTGTTTGATGATAAAAATAGACCGCATCATGGCA ATATCAATCGGGAATTAGATATTCTTGATTATCTGCTAGGGTATTTTGGTACACAACGTGCAAAGCGTCCATTA ACGCAGGATATTCATGATTTTCTGCAAACCCAGCGAGGTTATTTGGCACAGCAGCCGGAGTTGGGTTATCAG CGTGATAATATCCGTATTGATCGAGTTTCAGCTTTACAAAAACGTATAGCAGCCCGAATTGGGCTAGATGGTA CTATTTTCAAAGAATCGGTTGATTTAAGTAAGTTACCTTTTTATTTGATTGAACATCGTCAGCTTTTACCAAATT TACCCCATCTTGACTTTCAACATGATCAAACTCCCCAATCTTTTGTGATTTCCGACAACATTGTTAAAGTGAAA CAAGCGGGAATAGCAGATAAAATCGTTCGTGGACAGCTTATTGATTTTATAGATATTGAAAGCAAATTTACCG TTCGTGCCCAAATGATTGTCGCTGTAGAGGGAAATGAATTTTCTCTGGATACAAAAAATAGTATTCAACTTGAA AAGAATCTGCAGTTATTACAATCAGCGTCTGAGAAAAACAATTTACGATGGAGAAATAGCACGGCGTGGTTAG AGGATATGACGTATCGTATCAATTATACTGACGATCAGGTTATAGACGATAAAACAAAACAATGTCGTTTACAA AGTAATACTAAATCGCCTTTTCCAGCCTTAATTGCACCAAAAAATAAGATTACGATTATTAAGCAATCTTCTCC ACTCTCCAGTATTGCTGAATTTACTGATGAACCAGAATTCAAATTAGTTGCAACGGTGACAGAGATTGATCGG ATTGAAGGGATATTGACTATCGAACGGGATGACAACCAACTCCCTTTCCCGACTAAAGAAGAGAGTAATCAAT ATATATGGTACATATCTGATGAAAACTATATTTCAAGTGATCGTTTCTCTTTTGTGGTGAGCGTCGTGCTGAAT CGCGGTTTGGTTGAAAGGGAAGATATTGATCAATATAAGCTAGAGGAATGGATAGAGCGTGAAACACTTGCA GAGTTTCCTGCACATATTTCGTTAATTACTCATTGGCTGGCATCTGAAAATTTCGATGATTTTGCGAAGACATA TCAACGTTGGCAAAACAATGGGGCGCAGTTAGGGGATGAATCCTACACCATTTTGGAAAAACTGACATTAGG GCATTTACCAACAGGACTTACTGGCATTAGTAATATGTTTATTGCTACAGAAGCTCAGCGTCTAGAAGTTGTT GGCGAGAGTGGTAATGAGTGGAATACCCAGGCAATTATTAACAACGAACTATTCTATGTTCCCTCACAGAATA GTTAATACCGAGTGTTGTGATCAACTTTTATTATAAGCCGGAGGATAAATGGACAACAAAAATAACAAACCTAC TGATCAAGAGATTCTAAAAACATCACGGGCTGTCGGAGAAATTCCTTCAGCGGATAATTTAAAAAATCGTTTTA AAGCTCGTTCGATTCCATTAGAGACGGATTTTACTAATCTCATTGACCTTGCTGAAGTTGGACGATTGGCTAT CGGCCAGTCACCATCGCAGCAAAGTAAAACGCCTGGCACCGGAATGGAATTAACTTCGGATGGTAAATTACA AGTCAAGGCTGGGGCAGGTGTTGATATCGATAATAATAATCGTATTACTATTAAGTCTGGTCATGGAATTAAG GTTGATGGAAACGGCATTTCCGTTAAACCAGGTTCGGGTATTAAGGTTGATAGTAATGGTGTAAATGTCAATA TTGATGATTTTTGGGAGGAAATACGCAATAAAATTATGCCTAAAGGAACCATGCTGCCTATTTATGGCACACC TAACCCCTCTGCGCTGCCAACAGGATGGGAATGGTGTGATGGTAAAGATGGCAGACCTAATTTAAAAAAAGG GAAATATAACTTACTATCAGGTCAGTCTTCAGGTACTGATACTTTTTGGGCAGATAATAAGAATGGAGATACA GAGATCAACGTGTTATTTGTTTACTATATGATTAAGGTTGTGTAATATCTTAAGTAATATGCATTACTCTAAAAT GAATGATTTATATTTAAGTAACATAATAATTAAGTTGTGTTGTAGGGCTGTTTTTATGAGAAATATAAAAACGGA GGTAATAATTGGCTTCAAAATATCAGTGATGAAATAGAGTTATTTCGCTTTATAAAAATTTGTTTTATTTCTTTT AATAATTATTTATAGAAGGTAATGATATGTGCACACAAAAAAACGTGTTAGATAGACTGAAAGATAGAAATATT ACATTGGGTTGGGATGTTGTTGTTGCATATAACCAAGAAAGTGTTAATAAGTTATTGAAGCAACAATATGTTGA AAAAGTTTACTCAAATGAACATTTTGTTTTTAAAGATTGGCATGATGATAATAAAACGAAATTTATTGAGGGATT AACAGTAGGCGCTCCACTAGTTTCATTTGAGGAGGCGTCTTTATCCGATGCTAATGTAAAAGTGACACTTAAC TTTCTTTCTGGTAGATGGAGAGTTATACAAGCAAATACCGGCACACCAATTGAATGGAAAGAAATTGTTCCTG GCAGTGGCTATAAAGCAGAATTAGTTGTTCCGCTTAAATCAATAACTGGTAGTGTAAGTAAAAAAGATATCATA TTAAAATTCAAAGATGCTGTCGTAAAAAAAATAAATTTATTTGACAATCAAGAGCCTGATTTTATTAATTATTTC AAGCAATCGATCAGTGAGGGAAATTATACTTTAGGGCAACTGGTGACAGACAGCACACCGGGATTAATTCCT GCTGAATTTCATATTCGTACTCAACCCCATCCAAAAACACGTGAGCGTGGTTCTCAATATGTAGGAAATGGTG CGGTACTGTTGTTTATTAAAACGCAATATGGCGGAAGTGGAACATTGCCTGTAAATGATTTTGATTGGTTAATT CCTGATGATCATACTAGCGCATTAGTCATTTCGAGTAAGACCATGATGGGGCAAATATTGCCAAAACAATACA AAGATAAATTGCCTGGTGATCCTCAGTTTAGCCCACCAAAAAGAGTCAATGATAAACAAGACTCTGCTTATTAT ATTACGATTACCGATGGTGGATTTGATGGTAATAGCCCTATAGAGAAGTCATGGTTACGTTCTGATTATAGCA ATGGGATTTGGACTGGTGAACGTGGTAATGCTATTATTGGTGAAAAAGGAAAGCGGATACCACCACGTTTTC CATACCAAAATTTTGTTATTAAACCTCATGGTGAATCGTTATTTCAAGGATGGGAGAATAAGATAAATTACACT CAAAAGTGTGCAAGATATTTCCGACATCATAGTAATAGTATAACTTTCGAAGATACTGCATTAATGGATCTCAG TATTGGTGGACAAGGTAGTATCAATTGCCAGATTGATGGTGAACATTTCTATTTAAAATCAGATGATTTTTCCC CCAATGTCAGCTATGAACCAACTTCATTCTGGGATAAATTTATCGGTGGGGTGGATGCAAATGTGAAAGATGA ATTCAGAGATGAATTAGCACAACAGGCAGAAGCAAAGTTAAAACAGGTATTTAATATTGAATTGCCTGAAATC AGTCTGTTTTCTATTAAACATCTGCTCTTTCCTGGCATGGATGTTATGCAACTTAAACAGGGTTATTTCCCAGG AGATTTGATTATCTTTGGGGATATTTCACCTAAATTGACCACAATTCAGGTGGCTCCTTTGGAAGCCATGGTT GCCCTTAAAGAAAATCAAAAATTCACTGTCGTACCTGAAAATAAAAATGTTAGTTGGAAGTTGGATCATAATAG TGAGGCTATCAATGATCCGGGAAATATTGATGATAAAGGTATTTATACGGCACCGGGCAGAATCAGATCTGG TTCTGAAGTCATTAAAGTCACTGCAACTGACGGCGATGGAAATCAGGCATCGGCGGCGCTGACGTTGGTTCC TTCTTCTGTTGCATTAACACCTTCTTTTGCTTTTATCTCTGAAGCAGATAAGAAACCTATATTATTATTGGCGAA TGTCCTAGACGGAAAAGCAGTAACATGGAATGTGGAAAGCTGTACAGGCAGCCAATGTGGTTCTGTTGATCA GAATGGGCTTTATACTCCACCAGCAGGGCGTTTTAACGATGGATTTACTTTTGCATCCATCACCGCAACTGCA AAAGATGGTAGTCAAGCACGAACCATTATTTGTCTAATGGCATCAATGCCAGGACATGGTTTTTACAAGGTTG AACCTAATTTACGTTTGAATGTGAAAGTAGGGGAAGAAATTATCTTTAAAGCGCAGGCAGATAGCTATAATGG TGATCCTGATACTTGGGAAATTTTCCCTCCTCGCGGAAAATTAAGTGAACCTGAGTTTGAACCCAATAATGAT CCTGAAACTAATGATACAATTTTTGGTCATTATAAGGTGACCTATACCGCGCCGACTAATGTTACCTCACCTG AATTGCTTGTTGTCCATGTATGGGAGAAAAATAGGCATAATGAGAAAAACAAAGGTAAGGCAGGATATGCACT TATTGAAATTATCCCAGATGATAAATAGAAAATTTATTTAAATAAAAATCACAGCGGGTTTATCTCGCTGTGATT AAAGTCATCTTTTTTTATAGATTGTTTATCTCTAATAATAATTTTATTTTATAATATAAAGGAAATTAAAATGAATA ATGAATATAAAAATAACACCGTGAATTGGCGTATTTCACCTGATACGGTAGGAAGTATTGATAATAACGGTTTA TATACAGCACCTAATCGGGTAAAGAATATCGAATTTGTCCAAGTAATGGCAAGCGATGCTAATAATAATCAAT CTTCTGCGATTATTACTGTTATTCCCTCTTCTGTTGCGTTAACGCCATCGTTTACTTTTATCTCTGAGGCAAAA AAAACATCAGTCACTTTTAAAGCGACAGAACTTGAAGGGAAAAAAGTGACATGGAGTATAAATAATTATACCA GTAATCAGTATGGTTCCATCGATCAAAATGGTATCTACACACCACCGGAAAGTCGTTTTAACGATGGATATAC TTTTGTATCTATTACAGCAAAAGCGGAAAATGGCGCTGAAGCGCAAGCGCTTATTTGCTTGATGGCCAAAATT CCAGGGCATGCCTTTTTCGATGTTCAGCCTAATATATGTTTAAGTGTGAAGCCTGGAGAAGAAATCATTTTTA GAGCTAACGCAGATCGTTATAATGGTGATCCTGATTCCTGGGAAATTTTCCCGTCTCTTGGTAAATTGGGTGA GCCTGAGTATATAAAAAATAACGATCCAGAAATTCCTATTTATGGATATTATCAAGTGAAATATATTGCGCCAA CCAATATAAATTCTTCCCAAATACTCGTTGTGCGTACTTGGGAATATGACAAACATGATGAGCATAATCAAGGT AAAGCAGGATATGCATTCATTGAAATTGTGCCAGAAAATGAGCTTTAATATATATACCCAATAGATTTCGAGCC GCAGCGCGGCGGCAAGTGAATGAATCCCCAGGAGCATAGATAACGATGTGACTGGGGTGAGTGAACGCAG CCAACAAAGAGGTAACTTGAAAGATAATGAGTATAAATGACTTTAGTAAGAGAAATTATGGCTTCATTCAGAAC TATTTATTAGAGTAATTAACTTTATAAAGACATTTAATGGAAAATATAATAGAAAAATTTAATATTAATATTGAAG TCTCATCTGAAATTATTGGAGAGAGTTTATTAAACTCCCCTTTATTGATGAGTAGAGAAATCAGCAATCAATTA TCTGAAATATTATTAGATTATAAAGAATATAATATTGCATTGGATAAGTTAGTGTTAAATATAGGAGAAATACCC TATGAAATATTTGAACAACAATTCTATGGTCGTTTGGGAAAATTATTAAATGAAAAGTTAACAATAATAATAAAT GATAAATTATTGGTAAAAAACATATCAACCTCGTTATTTCCTGAATGTTTTAGTGAAAAAAGAAACCCATTATTA AATAGAGTCATAAAAAATTTACCTTCTAATTTGGTTTTTGAAGTTCATTCAATGGTAAAAATAGAATCAGTAAAT AACAAAAAACAAGCTAATATATTGACATCTTATCTGGCTTATTCTTTTTTTAATAAAAGCAAATTACAACAACAT TTATTTTCCACTAGTAATAATAAATTAATTGAGAGCTTATACGCACTTTTTCTAACGGATCAGAATCGAATACCT ACTGCTCATAAAATAGGAAAAGGTGCACTTATACTATCTGCCCTTATTTGGCTTTATTCTAATTCCAATGATTAT CTGCCCAAACCAGAAAGCACTCTGTTGTTACAAATAGAACAGGATATAAAACAAGGATATTTGCCTTTAACGT TGTTAATCACTTTCTTCCAGAACAGAAATGGCGGGCGTGTTTTTTGCGATTGGCAGTATGCGTTATGGCAAAT CGATATCATCAAAAATCACTTAGGCATTAAAATAACATCGAAAGAACCCCATTTACGGGAGAAAATAATGTTAC AACCAGTTAATGCTTCTGATCGATCCTCTGTGCTGATATCAGACGAAAAATTGACAATACCGTTAACAATTACA GGTGCGGGATTAGTGCTTCTCTGGCCACTATTAACTCCACTATTTTCGTCTTTTGATTTGTTAGATAAGAAAAG TTTTTCAGACAATTTGGCACAGGAAATAGCATTTAATTTATTGGAATGGTTAGTCTGGGGAGATGAGATGCTG TTACATCAGGAATCATCATTATCTTTATTACTCTGCGGAATAGATCACCAAACAATACTGGAGCGCCAGGTTCT TATTCCTGAGCACAAGGAAAAATTAAATAACTGGTTGCAAGGTATTTGTACTCAACTTTTCTCTTGGAAAAAGC TAGGGATCGATGATATGCGCCAACTTTTTTTGCAGCGTCAGGCTGCACTTTATTATGAAGATGATGGCCGGTTG GTTATTAACGGTGCAGCGTGAAGCTTATGATGTATTACTGACTCAAATGCCTTGGCCGTGGCCATTGAATATT GTGACATTACCTTGGCTAGCTGAGCCGATTAGTATCACTTGGGAAGGTATCTCTGAACCAACGGATTTGTCAT TTTGGTAATCCAATATCTCATTAGGAACTCTATGCATGTACGATTTATCTGATGATCTTGCCAGACAGAATATT TCACCGGAATATGAATTGACGGTTTTGCTGTCTCAGACTGCTATATTGGATAAACGAATTCGTTTACGAATTGA GGAATTAATGCAACAGCAAACAC;TATTGGGAGAAAGTGGACAGACGTCTTTTGATGATATTTGATTTTUATTC GTTTGGAGTGAAGAAGAAAAATGATGTTATTTGGTGTGAGGGGATGAAAATTGGAGGAAAGAGGATTTTGGTG CTGAGCCGATCCCATCTCGTAGCCGTCTAGGACAATTAGTTGAACGGTTTGACTTAACTCAATTTGAAATTGA TTTGATTTTATTGTGGGTGTTGGGTGATGTTGAGAGAGGTTATGTAAGGTTATTTTCTCTTGTTGGGGTAAGTG GAGGTAATAACAGCAAAAAGCAGATGTTAACGTTGGGATTGGCTTTGGAGTTGCTTTGTCCGAGTGTAGTAG AGCGCAATGCGCAACGTGCCAGTTTATTACCACAGGCACCGCTTTGGGATTATCGTTTATTTCAGTTGCGCG GTGATATGTCTGTTTCCTACGATGAAATACCGTTAGCAATCGATAATTCTCTTATGCATTGGTTATTGGGGCAT GATGGTGTGGGGATTTGTGTTGTGTGGGGGGGTGATTGGGTTGGTGTTGGTGAAGTGGGTGATATTTTGGGTG ATTTCACCAACCAATTGATAGAACTCTGCCAAATGGAACAAGAGGGGATGCTGACAATAATCGCCGGCGGAG CCGGAAGTGGCAGCAAAACAAGTGTTGCACGCGCAGCATCACAAGTAGGGCGCTCTGTATTGTTGTTATCGT TAGCATCAGTGACACTGAGTGAACATGAAACTATTACACTGATAACACTGGCATTACGTGAAGCACAACTAAG AAATGGGTGTGTTATGTTTGAAGGTTTGGATGAGTTTTGTGAAGGAGGGGGGGGTTTGGAGGTGTGGGTAGGA AATGGAGTGGGTGGTTGTTGGATTGGGGTGTTTTGTGAATTAGGTAAGGAAGGATGATTATTGGGATTGGATG CAATTTCACAAGTTGTATTGTCTATGCCAATGCCTTCTTTAATGGTGAAGGCTGCAGCATTAGCTTCAATGATG AGGAATTATTTTGGAGAGAATTGATTGGATGTTGAAAGTTTAGTGAGATGTTTGGATGGTTGTGGATTGATATTG AAAAAGGCCCTTAGTGAAGCAGAAATTTATCGCCGACTACGGGGGGAAACGGCTAGTTTGAGATTAGATGAT GTGGAAATGTGGGTGGGTTTTGGGTTAGAGGAGAATTTTGGAGGTTTAGGAGAGAGAATTAGAGGAGAAGGAA CCTTTGATGATTTGATCATCAGTGAATCTCAACAGCAACAATTACAAGAAATCCTGGCGGCTATTCGGCAACG AGATAGGATGGTAGAGGAAGGATTTGGTGGTAAAGTGAGGTATGGGAGGGGTATGAGGAGGGTATTTTTTGG TGAATGTGGGAGAGGAAAAAGGATGGTAGGAGAAGTGTTAGGTGGTGTTTTAGGTGTGGATTTGATGAAGGT AGATTTGTGGAGTGTGGTTAAGAAATATATTGGTGAAAGTGAAAAAAATGTGGGTGGTGTTTTTGATTATGGGG AAGAAGACGCCGGGGTATTGTTCTTTGATGAGGCAGATGCATTGTTTGGCAAACGAAGTGAAACTAAAGATG CAAAAGATCGTCATGCTAATATTGAAGTTTCCTACCTATTGCAACGCCTTGAAAGTTATCCAGGGCTGGTGAT ATTAGGGAGGAATTAGGGTAATGATTTAGAGTGAGGATTTAGTGGTGGGGTGAGTTTTTGGGTAGGATTGTGTT TTCCAGATGTTTCCTTACGGGAACGGATGTGGCGGATTATCTGGCCATCGGGAATTCAATTAGCCGACGACA TGAGTTTTTGAGGGTTGGGAAAAGGGGGTGAATTAAGGGGGGGGAATATGGGTAATATTGGGGTAGTGGGTA GTTGGCTGGCAGTAGATGAAGGAAATGAAAAAATTACTATGGCTCATATTGAATGCGCATTACGACGTGAACT GAGTAAAGTTGGGGGGATTGATTTAGGTTAATTTTTGTTTGTAATGGGGAGAGAAGTATGGTTAAAAATATGAA ATCAGATGAAACCTTACTGATATTAAATAGTAAAATAGAAGATGCATTAAAAGCGTATTTACCGGGCGAAGAT GTGGTTATTGGGTTGGATATGTTTGGTAAAAATGAAAATGGAGATTGTGGTAGGGTGTGGGTTTTTGTTTATGA TATTGAGGAAGATGTGGAATTAGGGGTGGGAGAAGGGGGGGAATAGGTGGGTGGGAGAGGAAATTTTGTGGG GGGATGTGTCAATGTTCGTTGTAATTATCTTATTTCCTACTGGGAGCCGGAACAGAGCGGAGGGCAGGGATC GCCAACCATACGTTCTAATAGTCAATCAATGAAGATAATGAACTGTGTATTGAATGCATTAATTAATCATCGTT CATTTCCTGGTTTACCCAGAACTTATACGAGAGTTCTTCCTCCTAATGAACAATTAAATAGCTTAGGAAACTTT TGGCAATCATTAGATAATAAGCCTCGACTATGTTTAAGTTATATGGTGACTATTCCTATTCAACTTACCCCGCC GAGAGAGAAGGTATGTGGTGTGATTAGGTGAAAAAGTGATATTAGTGGAAAAGGATGGGTTAAGTTTTATGTTG AGGCAGATGAAATTATCCGTCAGGCATTAGTTGATGCCTTAATATCTCAAACAACAGAATCTATGGATACGAT AACTAGCTGGCTGGCAAAAGTTGTTATTATTTGTCGACCACCAGAAATAATGAATAAACAAATGATTGAACAAA CTGTGAAATTAATTATTGCTGGAATTACAGAAGAGGGATTAGCTGGAAATATAAAGACAATCACTCAAAAGTG GGTGGAAGAGAAGACGATTATTGGTGAAATCGACGATGTTTCTCTAGTTATTTCCCAAGTTGACACGACAGC GTTGTCTGCTGTAACAATACCGACATCTGTTTAA (Pnf epitope) SEQ ID NO: 96 TGQKPGNNEWKTGR (PVCpromF) SEQ ID NO: 97 TATCATATGTCTACAACTCCAGAACAAATTGCTG (PVCpromR) SEQ ID NO: 98 ATCTCTAGAACAGATATTCCAGCCAGC (ParaINF) SEQ ID NO: 99 GGCGTCACACTTTGCTATG (ParaINF) SEQ ID NO: 100 TCGGTGGCAGTAAATTGTCC (F1 primer) SEQ ID NO: 101 ATGTCTACAAGTACATCTCAAATTGCG (F2 primer) SEQ ID NO: 102 GACTCCCTTGAGGGTACGG (F3 primer) SEQ ID NO: 103 TTCTGATGAGAGTGATGGTAC (F4 primer) SEQ ID NO: 104 TGAATAAAGAATTCAGTCAATATC (R1 primer) SEQ ID NO: 105 TAGTGGCTGATGAAAGTCTG (R2 primer) SEQ ID NO: 106 GGAAGCCAAAGATAATGAAGTG (R3 primer) SEQ ID NO: 107 CATTTCTTCCCTATGGTTG (R4 primer) SEQ ID NO: 108 TTAAATTCCTACAAGATTATCTTT (tBid amino acid sequence) SEQ ID NO: 109 RSSHSRLGRIEADSESQEDIIRNIARHLAQVGDSMDRSIPPGLVNGLALQLRNTSRSEEDRNRDLATAL EQLLQAYPRDMEKEKTMLVLALLLAKKVASHTPSLLRDVFHTTVNFINQNLRTYVRSLARNGMD (E. coli Sequence Optimised tBid bases) SEQ ID NO: 110 CGGTCAAGTCACTCGCGTCTGGGGAGAATCGAGGCTGATAGTGAGAGCCAAGAGGATATCATAA GAAACATAGCACGCCATTTGGCACAGGTAGGCGATTCTATGGATCGCTCCATCCCGCCTGGACTT GTCAATGGTCTTGCGCTTCAACTTCGTAACACTTCCCGGTCCGAGGAAGACAGAAATCGGGACCT TGCGACTGCTCTGGAACAACTGCTTCAAGCATATCCTCGTGACATGGAGAAAGAAAAGACTATGT TAGTATTAGCTCTTCHTTAGCTAAAAAGGTAGCTTCGCACACTCCAAGTTTATTGCGGGACGTTT TTCACACCACTGTTAATTTCATCAATCAGAACCTGCGTACTTATGTGAGATCTTTGGCGAGAAATG GTATGGAT (BaxBH3 peptide (aa59-73)) SEQ ID NO: 111 LSESLKRIGDELDSN (E. coli Sequence Optimised BaxBH3 bases) SEQ ID NO: 112 CTGTCGGAGAGTTTGAAGCGTATAGGTGACGAGCTGGACAGCAAT