CN114107232B - BALB/c mouse adapted strain of SARS related coronavirus WIV1-RsSHC014S and application thereof - Google Patents
BALB/c mouse adapted strain of SARS related coronavirus WIV1-RsSHC014S and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of coronavirus, in particular to a BALB/c mouse adapted strain of SARS-related coronavirus WIV1-RsSHC014S and application thereof, wherein the full-length gene sequence of the strain is shown as SEQ ID NO: 2, respectively. The mouse adapted strain can be well replicated and proliferated in lung tissues of wild BABL/c mice, so that the lung of the mice generates obvious pathological changes, the mice die, and respiratory diseases caused by SARS-related coronavirus can be well simulated to generate and develop processes. The research of the invention has higher reference practical value for the development of the medicine, the antibody and the vaccine of SARS-related coronavirus WIV1-RsSHC014S, and is more suitable for popularization.
Description
Technical Field
The invention relates to the technical field of coronavirus, in particular to a BALB/c mouse adapted strain of SARS-related coronavirus WIV1-RsSHC 014S.
Background
Coronaviruses belong phylogenetically to the order of the nested viruses (Nidovirales) the family of Coronaviridae (Coronaviridae) the genus coronaviruses (Coronavirus). Viruses of the genus coronaviruses are enveloped (envelope) RNA viruses with a linear single-stranded positive strand genome, and are a large group of viruses that are widely found in nature.
The coronavirus has the diameter of about 80-120 nm, the 5 'end of the genome has a methylated cap structure, the 3' end has a poly (A) tail, and the total length of the genome is about 27-32kb, so that the coronavirus is the virus with the largest genome in the currently known RNA viruses. Coronaviruses only infect vertebrates, such as humans, mice, pigs, cats, dogs, wolves, chickens, cows, birds. The coronavirus is firstly separated from the chicken in 1937, and the virus particles have the diameter of 60-200 nm, the average diameter of 100nm, are spherical or elliptical and have polymorphism. The virus has an envelope, spinous processes exist on the envelope, the whole virus is like coronas, and the spinous processes of different coronaviruses have obvious difference. Tubular inclusions are sometimes visible in coronavirus infected cells.
2019 the novel coronavirus (SARS-CoV-2, causing novel coronavirus pneumonia COVID-19) is the 7 th coronavirus which is known to infect human at present, and the other 6 are HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV (causing severe acute respiratory syndrome) and MERS-CoV (causing middle east respiratory syndrome), respectively.
The SARS-associated coronavirus RsSHC014 can effectively replicate and propagate in a corresponding cell line and a transgenic mouse by using human ACE2 as a receptor, and has certain cross-species spreading risk, but the existing neutralizing antibody and vaccine aiming at SARS-CoV or SARS-CoV-2 cannot provide effective protection effect for the strain, so that the research on pathogenesis of the strain and the storage of medicinal antibodies are very important in advance.
Animal models are important platforms for research on pathogenic mechanisms and drug development. RsSHC014 cannot effectively replicate and proliferate in a wild BALB/c mouse, a human ACE2 transgenic mouse is expensive and not easy to obtain, the transgenic mouse infected with the RsSHC014 can cause brain infection and death, and the pathological process and the generation and development mechanism of the human infected with coronavirus in lung tissues cannot be simulated, so that the wild mouse adapted strain is constructed for research and is important.
Disclosure of Invention
The invention aims to provide a BALB/c mouse adapted strain of SARS-related coronavirus WIV1-RsSHC014S and application thereof. The strain is characterized in that the genome contains a plurality of mutation sites different from the original strain, the BALB/c mice can be severely reduced in weight and even die due to virus infection, the respiratory tract has obvious virus proliferation, and pneumonia symptoms such as inflammatory infiltration, edema, cellulose exudation and the like can be caused in the lung.
In order to achieve the purpose, the invention provides the following technical scheme:
a SARS-associated coronavirus WIV1-RsSHC014S, its full-length gene sequence is shown as SEQ ID NO: 1 is shown.
The preparation method of the SARS-associated coronavirus WIV1-RsSHC014S comprises the following steps: (1) constructing a chimeric virus with RsSHC014 Spike gene on WIV1 as a framework, replacing the Spike gene of WIV1 with the RsSHC014 Spike gene, obtaining an infectious clone plasmid of WIV1-RsSHC014S by utilizing a BAC system, and constructing a chimeric virus genome sequence as shown in SEQ ID NO: 1 is shown in the specification; wherein, the coronavirus RsSHC014 is GenBank: KC 881005.1; coronavirus WIV1 is GenBank: KF 367457.1;
(2) virus rescue
And (2) taking 6 mu g of WIV1-RsSHC014S infectious clone plasmid obtained in the step (1), transfecting Vero E6 cells, collecting cell supernatant after 72 hours, freezing and thawing once, then diluting in a gradient manner, incubating new Vero E6 cells, culturing by using an agarose solid medium, and selecting one plaque for virus propagation after 5 days of culture to obtain the monoclonal chimeric virus WIV1-RsSHC 014S.
The invention also provides a BALB/c mouse adapted strain of SARS-related coronavirus WIV1-RsSHC014S, the full-length gene sequence of which is shown as SEQ ID NO: 2, respectively.
The preparation method of the BALB/c mouse adapted strain specifically comprises the following steps:
(1) mouse adapted strain passage
1) Selecting 3 wild BALB/C mice with 6-8 weeks old for infection, wherein the sex is not limited; the BALB/c mouse is easy to obtain and is applied to the construction of various coronavirus adaptive infection models, and related SARS-CoV and SARS-CoV-2 mouse adaptive strains can better simulate the development process of virus infection human bodies, so that the BALB/c mouse is selected to carry out RsSHC014 adaptive passage;
2) anaesthetizing the mouse with isoflurane, weighing, and carrying out anaesthesia by injecting alfuzin into the abdominal cavity at the concentration of 250 mg/kg;
3) the rescued WIV1-RsSHC014S was expressed as 1X 105 TCID5050ul each for nasal drop infection;
4) infected mice were kept on a constant rearing basis, and were weighed daily and observed for mental status. Dissecting after infection for 3 days, and grinding all lung tissues of the mice by using a grinding tube containing a DMEM culture medium;
5) grinding tissue at-80 deg.C with tissue grinder, centrifuging at 4 deg.C for 10min in 10000RCF centrifuge;
6) 3 mice lungs were mixed with 100ul of ground supernatants for infection 2 times, passaged to passage 19 in this manner, and steps 1) -6 repeated for each experiment);
(2) cell amplification and sequencing of mouse adapted strain
The mice have obvious pathological characteristics after being infected with 19 th generation virus, the 19 th generation lung is taken to grind the supernatant, the diluted supernatant is incubated with Vero E6 cells, and the cells are cultured in an incubator for 96 hours for virus amplification; and (3) taking cell supernatant, extracting virus nucleic acid by using a nucleic acid extraction kit, and performing second-generation high-throughput gene sequencing to obtain full-length genome information, such as SEQ ID NO: 2, respectively.
The amplified 19 virus generations were titrated at 1X 105 TCID50Each of the 3 BALB/c mice was infected by nasal drops at 50 ul. Body weight was recorded daily after infection and continuously observed until day 14 after infection, during which time lung tissue of dead mice was used for viral nucleic acid quantification and histopathological section analysis to obtain mice pathogenic strainsPhenotype.
The BALB/c mouse adapted strain of SARS-associated coronavirus WIV1-RsSHC014S of the present invention can be applied in neutralizing antibody, vaccine, detection reagent and related therapeutic medicine.
Compared with the prior art, the invention has the beneficial effects that:
the invention obtains the BALB/c mouse adapted strain of SARS-related coronavirus WIV1-RsSHC014S for the first time, the mouse adapted strain can be well replicated and proliferated in lung tissues of wild BALL/c mouse, the lung of the mouse is obviously changed pathologically, the mouse is dead, and the development process of respiratory diseases caused by SARS-related coronavirus can be well simulated. The research of the invention has higher reference practical value for the development of the medicine, the antibody and the vaccine of SARS-related coronavirus WIV1-RsSHC014S, and is more suitable for popularization.
Drawings
FIG. 1 shows the body weight change of mice during the generation of adapted strains;
FIG. 2 shows the body weight change trend of mice after P19 virus amplification and reinfection;
FIG. 3 shows the copy number of virus quantitative PCR of each tissue of mice after infection of P19 generation;
FIG. 4 is a pathological HE staining of lung tissue in mice after infection of P19 generation; wherein, P19-1 and P19-2 are 2 different viruses, and two different visual field images are selected after each virus infects the lung of a mouse to make a pathological section.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The viral sequences used in the examples below refer to the genome: bat coronavirus RsSHC014(GenBank: KC881005.1), bat coronavirus WIV1 (GenBank: KF 367457.1).
First, construction and rescue of rWIV1-RsSHC014S infectious clone
Constructing a chimeric virus with RsSHC014 Spike gene on WIV1 as a framework, replacing the Spike gene of WIV1 with the RsSHC014 Spike gene by using a reverse genetics technology, obtaining an infectious clone of WIV1-RsSHC014S by using a BAC system, and constructing the genomic sequence of the chimeric virus as shown in SEQ ID NO: 1 is shown.
The specific experimental process is as follows:
first, WIV1 viral cDNA was amplified: add 10. mu.l viral RNA, 20 ng hexa-base random primer, and add ddH2Supplementing O to 13.5 μ l, incubating at 70 deg.C for 5 min, and cooling on ice for 10 min; then 4. mu.l of 5 Xreverse transcription buffer, 200U M-MLV reverse transcriptase, 1mM dNTPs, 20U RNase inhibitor were added, incubated at 42 ℃ for 1h, at 75 ℃ for 15 min, and stored at-20 ℃.
The genome of rWIV1-RsSHC014S was divided into 8 cDNA fragments A-G (wherein the C fragment was divided into C1 and C2), RT-PCR amplified using the primers listed in Table 1 and cloned into the T vector. Viral cDNA was synthesized by reverse transcription using random primers. The KOD-plus kit is adopted for PCR, and the reaction system is as follows: mu.l of cDNA template, 1. mu.l of forward primer, 1. mu.l of reverse primer, 2. mu.l of MgSO4, 5. mu.l of 10 XBuffer, 5. mu.l of dNTP, 1. mu.l of KOD-plus polymerase, and finally ddH2O make up to 50. mu.l. The reaction procedure is as follows: pre-denaturation at 95 ℃ for 5 min; then carrying out 20 cycles of denaturation at 95 ℃ for 30 s, annealing at 60 ℃ for 30 s (0.5 ℃ per cycle) and extension at 68 ℃ for 5 min; performing 15 cycles of denaturation at 95 deg.C for 30 s, annealing at 50 deg.C for 30 s, and extension at 68 deg.C for 5 min; finally, extension is carried out for 10min at 68 ℃. After the PCR is finished, adding electrophoresis loading buffer solution into the product, carrying out electrophoresis and EB (Epstein-Barr) staining, and cutting a band with the corresponding size of each fragment. The nucleic acid fragments were purified using Gel Extraction Kit, the specific procedures being referred to the Kit instructions. In addition, to remove the Bgl I site at nt 1571 position, OE-PCR (overlap Extension PCR) was performed using primers FA, F-C1575a, R-C1575a and RA, and the synonymous mutation C1575A was introduced.
The amplified and recovered cDNA fragment was subjected to an A addition reaction and then cloned into a T load. Adding A to the reaction system: 1 μ l of 10 XTaq Buffer, 0.5 μ l dNTP, 0.3. mu.l Taq polymerase, 0.5. mu.l MgCl2And 8. mu.l of the recovered cDNA. The reaction was carried out at 70 ℃ for 30 min. The A-added fragment was inserted into the T-vector using T4 DNA ligase. A connection system: 0.5. mu.l of 10 XBuffer (NEB), 0.5. mu.l of pGEM-T EASY vector, 3.5. mu.l of A product, 0.5. mu. l T4 DNA ligase. Left at 16 ℃ overnight. Mu.l of the ligation product was taken, 60. mu.l of DH10B competent cells were electroporated, inoculated into 1 ml SOC preheated at 37 ℃ and cultured at 37 ℃ for 1 hour at 220 rpm, centrifuged at 800g for 2 min, 700. mu.l of the supernatant was discarded, resuspended cells were blown out with a pipette, and Amp + LB plates were plated with 10. mu. l X-gal and 20. mu.l of 1mM IPTG. Incubated at 37 ℃ overnight.
Table 1 (SEQ ID NO: 3-24)
| Primer name | Primer information | Function of |
| FA | tgaccgcggctaatacgatattaggtttttacctacccaggaaaag | Amplification of divided A fragments of WIV1 scaffold |
| F-c1575a | caggccatactggaattactggtgacaacgtggag | Removing the BglI enzyme cutting site in WIV1 skeleton |
| R-c1575a | gtcaccagtaattccagtatggcctgaaccaatatcagc | Removing the BglI enzyme cutting site in WIV1 skeleton |
| RA | atggtggccattatggctctaac | Amplification of divided A fragments of WIV1 scaffold |
| FB | tgttagagccataatggccaccatc | Amplification of B fragments of a partitioned WIV1 scaffold |
| RB | ctgtagccacgagggccctaagtttttccatagggac | Amplification of B fragments of a partitioned WIV1 scaffold |
| FC | cttagggccctcgtggctacagctcatagcgagc | Amplification of divided C fragments of WIV1 scaffold |
| RCm | acctgccgcctgggcagtttgtctgtc | Amplification of divided C fragments of WIV1 scaffold |
| FCm | ctgcccaggcggcaggtacagacac | Amplification of divided C fragments of WIV1 scaffold |
| RC | gctcataggcctcttgggcagtg | Amplification of divided C fragments of WIV1 scaffold |
| FD | cactgcccaagaggcctatgagc | Amplification of divided D fragments of WIV1 scaffold |
| RD | cagtgccaggtgggccctggagtgtagagtac | Amplification of divided D fragments of WIV1 scaffold |
| FE | agggcccacctggcactggtaagagtcattttgc | Amplification of E fragments of a compartmentalized WIV1 scaffold |
| R-EsBsaI | actggtctcttcgtttagttattaactaaaatatcactagacacc | The divided E fragment of WIV1 backbone was amplified and BsaI cleavage sites were introduced. |
| F-FsBsaI | tgaggtctccgaacttatggatttgtttatgag | The divided F fragment of WIV1 backbone was amplified and BsaI cleavage sites were introduced. |
| RF | aggtaggcctctagggcagctaac | Amplification of F fragments of a compartmentalized WIV1 scaffold |
| FG | tagctgccctagaggcctacctttc | Amplification of G fragments of a compartmentalized WIV1 scaffold |
| RG | actggcgcgcctttttttttttttttttttttttttgtcattctcctgagaagc | Amplification of G-sheets of a compartmentalized WIV1 scaffoldSegment of |
| F- SHC014 | agtggtctcaacgaacatgaaattgttagttttagtttttgctac | S gene for amplification of RsSHC014 |
| R- SHC014 | tcaggtctcagttcgtttatgtgtaatgtaatttgacacccttg | S gene for amplification of RsSHC014 |
Cleavage and enzymatic ligation of fragments: the pGEM-A is cut by Sac II enzyme, and the Bgl I enzyme is used for cutting after the CIAP treatment. The Sac II enzyme digestion system is 15 mug plasmid, 10 mug 10 XNEB CutSmart Buffer, 4 mug Sac II (20U/mug), and is complemented to 100 mug by ddH 2O; incubating for 3 h at 37 ℃; adding 1 mu l CIAP (30U/mu l), and digesting for 1h at 37 ℃. Then chloroform extraction and isopropanol precipitation: add 125 mu l ddH2And centrifuging for 2 min at the maximum rotating speed after shaking up and down for 2 min by using O, 50 mul 3M NaAc and 275 mul chloroform. Transferring supernatant, adding isopropanol of the same volume, mixing, standing in a greenhouse for 10min, and centrifuging at maximum speed for 10 min. Removing supernatant, adding 1 ml 70% ethanol, and centrifuging at maximum rotation speed for 5 min; and removing the supernatant, adding 1 ml of 95% ethanol, centrifuging for 5 min, removing the supernatant, centrifuging the residual ethanol to the bottom of the tube for a short time, and sucking and removing the residual ethanol by using a 10 mu l gun head. The pellet was air dried with the tube lid open. Finally, the precipitate was dissolved with a preheated Elution Buffer. Then 5 mul Bgl I and 10 mul 10 XNEB buffer 3 are added, and the enzyme digestion is carried out for 2 h at 37 ℃. The enzyme pGEM-G is cut by Asc I, and after CIAP treatment, the enzyme is cut by Bgl I. The Asc I enzyme digestion system is 15 mug plasmid, 10 mug 10 XNEB CutSmart Buffer, 4 mug Asc I (10U/mug), and ddH2O is used for complementing to 100 mug; incubate at 37 ℃ for 3 h. Then CIAP treatment, chloroform extraction, isopropanol precipitation and Bgl I enzyme digestion are carried out, and the method is the same as the above.BsaI orBsmThe BI enzyme cleaves the S gene. The enzyme digestion system is 15 mug plasmid, 5 mug 10 XNEB Buffer 3, 3 mugBsaI orBsmBI (10U/μ l) with ddH2O complement to 50 μ l, incubate for 6h at 37 ℃ (if used)BsaI) Or incubation at 55 ℃ for 3 h (if used)BsmBI). The pBAC-CMV vector was then digested with Sac II and Asc I. The enzyme digestion system is a plasmid of 5 mug, 5 mug 10 XNEB CutSmart Buffer, 1 mug Sac II (20U/mug), 1 mug Asc I (10U/mug), complemented to 50 mug by ddH2O, and incubated for 6h at 37 ℃. And finally, carrying out electrophoresis and recovering the enzyme-digested vector and all fragments, and then connecting and transforming the Escherichia coli. 1 XTAE electrophoresis buffer (stored at 4 ℃) containing 0.5 mM cytidine was prepared. Preparing 1% agarose Gel by using the buffer solution, carrying out electrophoresis on all enzyme digestion products, separating nucleic acid fragments with different sizes, cutting bands of the BAC vector and the fragments with the corresponding sizes, purifying by using Gel Extraction Kit, finally eluting the nucleic acid by ddH2O, and extracting once by using chloroform with the same volume. The specific operation is the same as that described above. The linker between the A-G fragment and pBAC-CMV is: 4 mul BAC vectors (enzyme digestion of Sac II and Asc I), 0.5 mul each of A-G fragments, 1 mul 10 Xbuffer, 1 mul l T4 DNA ligase (10U/mul), and standing overnight at 4 ℃. 1.8 μ l of the electrically transformed DH10B competent cells were plated on Chl + LB plates.
And (3) positive clone identification: positive clones were identified by multiplex PCR. Multiplex PCR primers were designed (as shown in Table 2) to amplify the junction sequences between the fragments and the vector. Each Primer was diluted to 10. mu.M and then mixed in equal volumes into one tube of Primer Mix. The colony PCR system is as follows: 2 XTaq master Mix 7.5. mu.l, Primer Mix 1. mu.l, ddH2O 6.5.5. mu.l, colonies. The reaction procedure is as follows: pre-denaturation at 95 ℃ for 5 min; then 35 cycles of denaturation at 95 ℃ for 30 s, annealing at 57 ℃ for 30 s, and extension at 72 ℃ for 1 min are carried out; extension at 72 ℃ for 10 min. 7. mu.l of the PCR product was subjected to electrophoretic analysis.
TABLE 2 multiplex PCR primers (sequences as shown in SEQ ID NO: 25-38)
| Primer name | Primer information |
| F1 | aacaactccgccccattgacgc |
| R1 | actgcaacacggacgaaaccgt |
| F2 | ccctggacaaggatgtgctggtt |
| R2 | cgacgggactctccaaagtgtggt |
| F3 | tggcgcgcttcatctctactttg |
| R3 | gccaagatctctaggcgtcatgttt |
| F4 | tgcatcacatggagcttccaacagg |
| R4 | acagcagcatacagccatgcca |
| F5 | tgctatccatgcagggtgctgta |
| R5 | tggaacacaaccatcacgcgca |
| F6 | gggaggttggaaaacctagaccacca |
| R6 | ctttggtcagcaatgtgcgcgg |
| F7 | agccaccacattttcaccgaggc |
| R7 | agaaggcacagctcccttagcca |
Virus rescue: vero E6 cells were transferred to 6-well plates one day in advance and 6. mu.g of pBAC-CMV-rWIV1-RsSHC014S plasmid was transfected into 1 well using Lipofectamine 3000. The empty pBAC-CMV plasmid was also transfected as a control. Obvious cytopathic effect can be seen after continuous observation for 3-5 days, and the recombinant virus can be successfully rescued.
Second, mouse adapted strain passage
1) Selecting 3 wild BALB/C mice with 6-8 weeks old for infection, wherein the sex is not limited;
2) anaesthetizing the mouse with isoflurane, weighing, and carrying out anaesthesia by injecting alfuzin into the abdominal cavity at the concentration of 250 mg/kg;
3) the rescued WIV1-RsSHC014S was expressed as 1X 105 TCID5050ul each for nasal drop infection;
4) infected mice were kept on a constant rearing basis, and were weighed daily and observed for mental status. Dissecting after infection for 3 days, and grinding all lung tissues of the mice by using a grinding tube containing a DMEM culture medium;
5) grinding tissue at-80 deg.C with tissue grinder, centrifuging at 4 deg.C for 10min in 10000RCF centrifuge;
6) the 3 mice lung mill supernatants were mixed at 100ul and passaged to passage 19 for the 2 nd infection, with steps 1-6 repeated for each experiment. The change in body weight of mice during the adapted strain passage is shown in FIG. 1.
Cell amplification, sequencing and pathogenic phenotype identification of mouse adapted strain
The mice have obvious pathological characteristics after being infected with 19 th generation virus, 2 mice lungs of the 19 th generation are respectively taken to be ground into supernatant, diluted and incubated with Vero E6 cells, the cells are cultured in an incubator for 72 hours to carry out virus amplification, and the amplified viruses are respectively named as P19-1 and P19-2; and (3) taking the cell supernatant to extract virus nucleic acid by using a nucleic acid extraction kit, and performing second-generation high-throughput gene sequencing to obtain full-length genome information, such as SEQ ID NO: 2, respectively.
The titer of the amplified 19 th generation two strains of virus is determined according to 1 × 105 TCID50Each of the 4 BALB/c mice was infected by nasal drops at 50 ul. Body weights were recorded daily after infection and continuously observed until day 14 after infection, during which time lung tissue from dead mice was used for viral nucleic acid quantification and histopathological section analysis to obtain the pathogenic phenotype of the acclimated strain on the mice. Wherein, the weight change trend of the mice after P19 generation virus amplification and reinfection is shown in figure 2; the copy number of virus quantitative PCR of each tissue of mice after infection of P19 generation is shown in FIG. 3; pathological HE staining of lung tissue in mice after infection in P19 passages is shown in fig. 4.
According to the experimental results, the mouse adapted strain can be well replicated and proliferated in lung tissues of wild BABL/c mice, so that the lung of the mice generates obvious pathological changes, the mice die, and the development process of respiratory diseases caused by SARS-related coronavirus can be well simulated. The research of the invention has higher reference practical value for the development of drugs, antibodies and vaccines of SARS-related coronavirus WIV1-RsSHC 014S.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Sequence listing
<110> Nanjing Keguan biomedical science and technology Co., Ltd
<120> BALB/c mouse adapted strain of SARS related coronavirus WIV1-RsSHC014S and application thereof
<160> 38
<170> SIPOSequenceListing 1.0
<210> 1
<211> 27311
<212> DNA
<213> WIV1-RsSHC014S(Artificial Sequence)
<400> 1
atggagagcc ttgttcttgg tgtcaacgag aaaacacacg tccaactcag tttgcctgtt 60
cttcaggtta gagacgtgct agtgcgtggc ttcggggact ctgtggaaga ggccctatcg 120
gaggcacgtg aacaccttaa aaatggcact tgtggtttag tagagctgga aaaaggcgta 180
ctgccccagc ttgaacagcc ctatgtgttc attaaacgtt ctgatgcctt aagcaccaat 240
cacggccaca aggtcgttga gttggttgca gaattggacg gcattcagta cggtcgtagc 300
ggtataactc tgggagtgct cgtgccacat gtgggcgaaa ccccaatcgc ataccgcaat 360
gttcttcttc gtaagaacgg taataaggga gccggtggcc atagctttgg catcgatcta 420
aagtcttatg acttaggtga cgagcttggc actgatccca ttgaagatta tgaacaaaac 480
tggaacacta agcatggcag tggtgtactc cgtgaactca ctcgcgagct caatggaggt 540
gcattcactc gctatgtcga taacaacttt tgtggcccag atgggtaccc tcttgattgc 600
attaaagatt ttctcgctcg cgcgggcaag tcaatgtgca ctctttctga acaacttgat 660
tacatcgagt cgaagagagg tgtctactgc tgccgtgacc atgagcatga agttgcttgg 720
ttcactgagc gctctgataa gagctatgag catcagacac ccttcgaaat taagagtgcc 780
aagaaatttg acaccttcaa aggggaatgc ccaaagtttg tatttcctct cgattcaaaa 840
gtcaaagtca ttcaaccacg tgttgaaaag aaaaagactg aaggtttcat ggggcgcata 900
cgctctgtgt accctgttgc atctccacag gagtgtaaca acatgcactt gtctaccttg 960
atgaaatgta atcattgcga tgaagtttca tggcagacgt gcgattttct gaaagccact 1020
tgtgaacatt gtggcactga aaactcagtc acagaaggac ctactacatg tgggtaccta 1080
cctactaatg ctgtagtgaa aatgccatgt cccgcctgtc aagacccgga gattggacct 1140
gagcatagtg tcgcagacta tcacaaccac tcaaacattg aaactcgact ccgcaaagga 1200
ggtaggacta gatgttttgg aggctgtgtg tttgcctatg tcggctgcta caacaagcgt 1260
gcctactggg ttcctcgtgc tagtgctgat attggttcag gccatactgg aattactggt 1320
gacaacgtgg agaccttgaa tgaggatctc cttgagatac tgagtcgtga acgtgttaat 1380
attaacattg ttggcgattt tcagttgaat gaagaggtgg ccatcatttt ggcatctttt 1440
tctgcttcta caagtgcctt tattgacact ataaggagtc ttgattacaa gtctttcaaa 1500
gccattgttg agtcctgcgg taactacaaa gttaccaagg gaaagcccgt aaaaggtgct 1560
tggaacattg gacaacagag atcagtttta acaccactgt gtggtttccc ctcacaggct 1620
gctggtgtta tcagatcaat ttttgcacgc acacttgatg cagcaaacca ctcaattcct 1680
gatttgcaaa gagcagctgt caccatactt gatggtattt ctgaacagtc attacgtctt 1740
gtcgacgtca tggtttatac ctcagacctg cttaccaaca gtgtcatcat tatggcatat 1800
gtaactggtg gtcttgtaca acagacttct cagtggttgt ctaatttgtt gggcactact 1860
gttgaaaaac tcaggcccat ctttgtgtgg attgaggcga aacttagtgc aggagttgaa 1920
tttctcaagg atgcttggga gattctcaaa tttctcatta caggtgtttt tgacatcgtc 1980
aagggtcaaa tacaggttgc ttcagataac atcaagggtt gtgtaaaatg cttcattgat 2040
gttgttaata aagcactcga aatgtgcatt gaccaagtca ctatcgctgg cactaagttg 2100
cgatcactca acttaggtga agtcttcatc gctcaaagca agggacttta ccgtcagtgt 2160
atacgtggca aggaacagct gcaactactt atgcctctta aggcaccaaa agaagtcacc 2220
tttcttgaag gtgattcaca tgacacagta cttacctctg aggaggttgt tctcaaaaac 2280
ggtgaactcg aggcactcga gacgcctgtt gatagcttca caaatggagc tgtagttggc 2340
acaccagttt gtgtaaatgg cctcatgctc ttagagatca aggacaaaga acaatactgc 2400
gcattgtctc ctggtttact ggctacaaac aatgtctttc gcctaaaagg gggtgcacca 2460
actaaaggtg taacctttgg agaagatact gttttggaag ttcaaggtta caagaatgtg 2520
agaatcacat ttgagcttga tgaacgtgtt gacaaagtgc ttaatgaaaa gtgctctgtc 2580
tacactgttg aatccggtac cgaagttact gagtttgcat gtgttgtagc agaggctgtt 2640
gtgaagactt tacaaccagt ttctgatctt cttaccaaca tgggtattga tcttgatgaa 2700
tggagtgtgg ctacattcta tttgtttgat gatgctggtg aagaaaaact ttcttcacgt 2760
atgtactgtt ccttttatcc tcctgatgat gaggaggatt gtgatgagta tgatgaagaa 2820
gaggaagtcc tggaagaatc ctgtgcgcat gaatacggta cagaagaaga ttaccaaggt 2880
ctttcactgg aatttggtgc ctcaactgaa atgcaagttg aagaagaaga agaagaggac 2940
tggcttggtg atgctactga attatcggag catgaaccag aaccagaact aacacctgaa 3000
gaaccagtta accagtttac tggttattta aaacttactg acaatgttgc cattaagtgt 3060
gtggacatcg tgaaggaggc gcaaaacgct aaccccacgg tgattgtaaa tgctgctaac 3120
atacatctga aacatggtgg tggtgtagca ggtgcactca acaaggcaac caacggtgcc 3180
atgcaaaaag agagcgatga ttacattaag ctaaatggtc ctctcgtagt tggaggttca 3240
tgtttgcttt ctggacataa tcttgctaag aagtgtctgc atgttgttgg acctaaccta 3300
aatgcaggtg aggacatcca gcttcttaag gcagcatatg aaaattttaa ttcacaggac 3360
actttacttg caccattgtt gtcagcaggc atatttggtg ctaaaccact tcagtcttta 3420
caagtgtgcg tgcagacagt tcgtacacag gtttacattg cagtcaatga caaagctctt 3480
tatgagcagg ttgtcatgga ttaccttgat agcctgaagc ctagagtgga agcacctaaa 3540
caagaggagc caccaaggac agaagatcct aaaattgagg agaaatctgt cgtacagaag 3600
cctatcgatg tgaagccaaa aattaaggct tgcattgatg aggttaccac aacactggaa 3660
gaaactaagt ttcttaccaa taagttactc ttgtttgctg acatcaatgg taagctttac 3720
catgattctc acaacatgct tagaggtgaa gatatgtctt tccttgagaa ggatgcacct 3780
tacgtggtag gtgatgttat cactagtggt gatatcactt gtgttgtaat accctccaaa 3840
aaggctggtg gcactacaga gatgctctca agagctttga agaaagtgcc agttgatgag 3900
tatataacca cataccctgg acaaggatgt gctggttata cacttgagga agctaagact 3960
gctcttaaga aatgcaaatc tgcattttac gtgttacctt cagaaacacc taatgctaag 4020
gaagagattc taggaactgt gtcctggaat ttgagagaaa tgcttgctca tgctgaagag 4080
acaagaaaat taatgcctat atgcatggat gttagagcca taatggccac catccaacgc 4140
aagtacaaag gaattaaagt tcaagaaggc attgttgact atggagtccg attcttcttt 4200
tatactagta aagagcctgt agcttctatc attacgaagc tgaactctct aaatgagcca 4260
cttgtcacaa tgccaattgg ttatgtgaca catggtttta atcttgaaga ggctgcgcgc 4320
tgtatgcgtt ctcttaaagc tcctgccgta gtgtcagtat catcaccaga tgctgttact 4380
acatataatg gatacctcac ttcgtcatca aagacatctg aggagcactt tgtggaaaca 4440
gtttctttgg ctggttctta cagagattgg tactattcag gacagcgtac agagttaggt 4500
gttgaatttc ttaagcgtgg tgacaaaatt gtgtaccaca ctttggagag tcccgtcgag 4560
tttcatcttg acggtgaggt tcttccactt gacaaactaa agagtctttt atccctacgg 4620
gaggttaaga ctataaaagt gttcacaact gtggacaaca ctaatctcca cacacagctt 4680
gtggatatgt ctatgacata tggacagcag tttggtccaa catacttgga tggtgctgat 4740
gttacaaaaa ttaaacctca tgtaaatcat gagggtaaga ctttctttgt actacctagt 4800
gatgacacac tacgtagtga agctttcgag tactaccaca ctcttgatga gagttttctt 4860
ggtaggtaca tgtctgcttt aaaccacaca aagaaatgga aatttcctca agttggtggt 4920
ttaacttcaa ttaaatgggc tgataacaat tgttatttgt ctagtgtttt attagcactt 4980
caacagattg aagtcaaatt caatgcacca gcacttcaag aggcttatta tagagcccgt 5040
gctggtgatg ctgctaactt ttgtgcactc atacttgctt acagtaataa aactgttggc 5100
gagctcggtg atgtcagaga aactatgacc catcttctac agcatgctaa tttggaatcc 5160
gcaaagcgag ttcttaatgt ggtgtgtaaa cattgcggtc agaaaactac caccttaacg 5220
ggtgtagaag ccgtgatgta catgggtact ctatcttatg ataatcttaa gacaggtgtt 5280
tccattccat gtgtgtgtgg tcgcgatgct acacaatatc tagtacaaca agagtcttct 5340
tttgttatga tgtctgcacc acctgctgaa tataaattac agcaaggtac atttttatgt 5400
gcgaatgagt acactggtaa ctatcagtgt ggtcattaca ctcatataac tgctaaggag 5460
accctctatc gtattgatgg agctcacctt acaaagatgt cagagtacaa aggaccagtg 5520
actgacgttt tctataagga aacatcttac actacaacca tcaagcctgt gtcgtataaa 5580
ctcgatggag ttacttacac agagattgaa ccaaaattgg atgggtatta taaaaaggat 5640
aatgcttact atacagagca gcctatagac cttgtaccaa ctcaaccact accaaatgcg 5700
agttttgaca atttcaaact cacatgttct aatacaaaat ttgctgatga tttaaatcaa 5760
atgacaggct tcacaaagcc agcttcacga gagttatatg tcacattctt tccagacttg 5820
aatggcgatg tagtggctat tgactataga cactattcag cgagtttcaa gaaaggtgct 5880
aaattactgc ataagccaat tgtttggcat atcaatcagg ctacaaccaa gacaacgttt 5940
aaaccaaaca cttggtgttt acgttgtctt tggagtacaa agccagtaga tacttcaaat 6000
tcatttgaag ttctggcagt agaagacaca caaggaatgg acaatcttgc ttgtgaaagt 6060
caacaaccca cctctgaaga agtagtggaa aatcctacca tacagaagga agtcatagag 6120
tgtgacgtga aaactactga agttgtaggc aatgtcatac ttaaaccatc agatgaaggt 6180
gttaaagtaa cacaagagtt ggatcatgag gatcttatgg ctgcttatgt ggaaaataca 6240
agcattacca ttaagaaacc taatgagctt tcactagcct taggtttaaa aacaattgcc 6300
actcatggta ttgctgcaat taatagtgtg ccttggagta aaattttggc ttatgtcaaa 6360
ccattcttag gacaagcagc aattacaaca tcaaattgcg ctaagagatt agcacaacgt 6420
gtgtttaaca attatatgcc ttatgtgctt acactattgt ttcaattgtg tacttttaca 6480
aaaagtacaa attctagaat tagagcttca ctacctacga ctattgctaa aaatagtgtt 6540
aggggtgttg ctagattatg tttggatgct ggcattaatt atgtaaagtc acccaaattt 6600
tctaaattgt tcactattgc aatgtggcta ttattgttaa gcatttgctt aggttcacta 6660
atctatgtaa ctgcagcttt aggtgtatta ttgtccaact ttggagctcc ttcttattgt 6720
agtggcgtta gagaatcgta cctcaattcc tctaatgtta ctactatgga tttctgtgaa 6780
ggttcttttc cttgcagtgt ttgtttaagt ggattagact cgcttgattc ctatccagct 6840
cttgaaacca tccaggtgac gatctcatcg tacaagctag acttgacaat tttaggtctg 6900
gctgctgagt ggtttttggc atatatgttg tttacaaaat tcttttattt actaggtctt 6960
tcagctataa tgcaggtgtt ctttggctat tttgctagtc atttcatcag caattcttgg 7020
ctcatgtggt ttatcattag cattgtacaa atggcacccg tttctgcaat ggttaggatg 7080
tacatcttct ttgcttcttt ttactacata tggaagagct atgttcatat tatggatggt 7140
tgtacctctt cgacttgcat gatgtgctat aagcgcaatc gtgccacacg cgttgagtgt 7200
acaactattg ttaatggcat gaagagatct ttctatgtct atgcaaatgg aggccgtggc 7260
ttctgcaaga ctcacaattg gaattgtctc aattgtgaca cattttgcac tggtagtaca 7320
ttcattagtg atgaagttgc tcgtgatttg tcactccagt ttaaaagacc aattaaccct 7380
actgaccagt catcgtatat tgttgatagt gttgctgtta aaaatggcgc gcttcatctc 7440
tactttgaca aggctggtca aaagacttat gagagacacc cactctccca ttttatcaat 7500
ttagacaatt tgagagctaa caacactaaa ggttcactac ctattaatgt catagtcttt 7560
gatggcaagt ccaaatgcga cgagtctgct gctaggtctg catctgtgta ctacagtcag 7620
ctaatgtgcc aacctattct gttgcttgac caagctctca tatcagatgt tggagatagt 7680
actgaagttt ctgttaagat gtttgatgct tatgtcgaca ccttttcagc aacttttagt 7740
gtccctatgg aaaaacttag ggccctcgtg gctacagctc atagcgagct ggcaaagggt 7800
gtagctttag atggtgtcct ttctacattt gtgtcagctg ctcgtcaagg tgttgttgat 7860
actgatgttg acacaaagga tgtcattgaa tgtctcaaac tttcacatca ctccgacttg 7920
gaagtgacag gtgacagttg taacaacttc atgctcacct ataacaaagt tgaaaacatg 7980
acgcctagag atcttggcgc atgtattgat tgtaatgcaa ggcatattaa tgctcaagta 8040
gcaaaaagtc acaatgtttc actcatctgg aatgtaaaag actatatgtc tttatctgaa 8100
cagctgcgta aacaaattcg tagtgctgct aagaagaaca acataccttt tagactaact 8160
tgtgctacaa ctagacaggt tgtcaatgtc ataactacta aaatctcact caagggtggt 8220
aagattgtta gtacttggtt taaacttatg cttaaggcca cattattgtg cgtcattgct 8280
acattggtct gttacatcgt tatgccagta catacattgt ctgttcatga tggttacaca 8340
aatgaaatca ttggttacaa agccattcag gatggtgtca ctcgtgacat cgtttctact 8400
gatgattgtt ttgcaaacaa acatgctggt tttgactcat ggcttagcca gcgtggtggt 8460
tcatacaaaa atgacaaaag ctgccctgta gtagctgcta tcattacaag agagattggt 8520
ttcatagtgc ctggcttacc aggtactgtg ttgagagcaa tcaatggtga cttcttgcat 8580
tttctacctc gtgtctttag tgctgttggc aacatttgct acacaccttc taaactcatt 8640
gagtatagtg attttgctac ctctgcttgc gttcttgctg ctgagtgtac aatttttaag 8700
gatgctatgg gcaaacctgt gccatattgt tatgacacta atttgctaga gggttctatt 8760
tcttatagtg agcttcgtcc agacactcgt tatgtcctta tggatggttc catcatacag 8820
tttcctaaca cttacctgga gggttctgtt agagtagtaa caacttttga tgctgagtac 8880
tgtagacatg gtacatgtga aagatcagaa gctggtattt gcttatctac cagtggtaga 8940
tgggttctta acaatgaaca ttatagagct ctacctggag tattttgtgg tgttgatgca 9000
atgaatctta tagcaaacat ctttactccc cttgtgcaac ctgtgggtgc tttagatgtg 9060
tctgcttcag tagtggctgg tggtattatt gccatattgg tgacttgtgc tgcctactac 9120
tttatgaaat tcagacgtgc ttttggtgag tacaaccatg ttgttgctgc taatgcactt 9180
ttgtttttga tgtctttcac tatactctgt ctggcaccag cttatagctt tttgccagga 9240
gtctactcag tcttttactt gtacttgaca ttctatttca ctaatgatgt ttcgttcttg 9300
gctcaccttc aatggtttgc catgttttct cctattgtgc ctttttggat aacagcaatc 9360
tatgtattct gtatttctct gaagcactgc cattggttct ttaacaacta tcttaggaaa 9420
agagtcatgt ttaatggagt tacatttagt accttcgagg aggctgcttt gtgtaccttt 9480
ttgctcaata aggaaatgta cctaaaattg cgtagtgaga cactgttgcc acttacacag 9540
tacaacaggt atcttgctct atataacaag tacaagtatt tcagtggagc cttagatact 9600
accagctatc gtgaagcagc ttgctgccac ttagcaaagg ctctaaatga ctttagcaat 9660
tcaggtgctg atgttctcta ccaaccacca cagacatcaa tcacttctgc tgttctgcag 9720
agtggtttta ggaaaatggc attcccatca ggcaaagttg aagggtgcat ggtacaagta 9780
acctgtggaa ctacaactct taatggattg tggttagatg acacagtata ctgtccaaga 9840
catgtcattt gcacagcaga agacatgctt aatcctaact atgaagatct gctcattcgc 9900
aaatccaacc atagcttcct tgttcaggct ggcaatgtac aactccgagt tatcggccat 9960
tctatgcaaa attgtctgct taggcttaaa gttgatacct ctaaccctaa gacacccaag 10020
tataaatttg tccgtattca acctggtcaa acattttcag ttctagcatg ctacaatggt 10080
tcaccatctg gtgtttatca gtgtgccatg agacctaacc ataccattaa aggttctttc 10140
cttaatggat catgtggtag tgttggtttt aacattgatt atgattgcgt gtctttctgc 10200
tatatgcatc acatggagct tccaacagga gtacacgctg gtactgactt agaaggtaaa 10260
ttctatggtc catttattga cagacaaact gcccaggcgg caggtacaga cacaaccata 10320
acattaaatg ttttggcatg gctgtatgct gctgttatca atggtgatag gtggtttctt 10380
aatagattca ccactacttt gaatgacttt aaccttgtgg caatgaagta caactatgaa 10440
cctttgacac aagatcatgt tgacatattg ggacctcttt ctgctcaaac gggaattgct 10500
gtcttagata tgtgtgctgc tttgaaagag ctgctgcaga atggtatgaa tggtcgtact 10560
atccttggta gcactatttt agaagatgag tttacaccat ttgatgttgt tagacaatgc 10620
tctggtgtta ccttccaagg taagttcaag aaaattgtta agggcactca tcattggatg 10680
cttttaactt tcttgacatc actattgatt cttgtccaaa gtactcagtg gtcactgttt 10740
ttctttgttt acgagaatgc tttcttgcca tttactcttg gtattatggc aattgctgca 10800
tgtgctatgc tgcttgttaa gcataagcac gcattcttgt gcttgtttct gttaccttct 10860
cttgcaacag ttgcttactt taatatggtc tacatgcctg ccagctgggt gatgcgtatt 10920
atgacatggt ttgaattggc tgacactagc ttgtctggtt atcggcttaa ggactgtgtt 10980
atgtatgctt cagctttagt tttgcttatt ctcatgacag ctcgcactgt ttatgatgat 11040
gctgctagac gtgtttggac actgatgaat gtcattacac ttgtttacaa agtctactat 11100
ggtaatgctt tagatcaagc tatttccatg tgggccttag ttatttctgt aacctctaac 11160
tattctggtg tcgtcacgac tatcatgttt ttagctagag ctatagtgtt tgtgtgtgtt 11220
gagtattacc cattgttatt tattactggc aacaccttac agtgtatcat gcttgtttat 11280
tgtttcttag gctattgttg ctgctgctat tttggccttt tctgtttact caaccgttac 11340
ttcaggctta ctcttggtgt ttatgactat ttggtctcta cacaagaatt taggtatatg 11400
aactcccagg ggcttttgcc tcctaagagt agtattgatg ctttcaagct taacattaag 11460
ttgttgggta ttggaggtaa accatgtatc aaggttgcta ctgtacagtc taaaatgtct 11520
gacgtaaagt gcacatctgt ggtactgctc tcggttcttc aacaacttag agtagagtca 11580
tcttctaaat tgtgggcaca atgtgtacaa ctccacaatg atattcttct tgcaaaagac 11640
acaactgaag ctttcgagaa gatggtttct cttttgtctg ttttgctatc catgcagggt 11700
gctgtagaca ttaacaagtt gtgcgaggaa atgctcgaca accgtgctac tcttcaggct 11760
attgcttcag aatttagttc tttaccatca tatgccgctt atgccactgc ccaagaggcc 11820
tatgagcagg ctgtagctaa tggtgattct gaagtcgttc tcaaaaagtt aaagaaatct 11880
ttgaatgtgg ctaaatctga gtttgaccgt gatgctgcca tgcaacgcaa gttggaaaag 11940
atggcagatc aggctatgac ccaaatgtac aaacaggcaa gatctgagga caagagggca 12000
aaagtaacta gtgctatgca aacaatgctt ttcactatgc ttaggaagct tgataatgat 12060
gcacttaaca acattatcaa caatgcgcgt gatggttgtg ttccactcaa catcatacca 12120
ttgactacag cagccaaact tatggttgtt gtccctgatt atggtaccta caagaacact 12180
tgtgatggta acacctttac atatgcatct gcactctggg aaatccagca agttgttgat 12240
gcagatagta agattgttca acttagtgaa attaacatgg ataattcacc aaatttggct 12300
tggcctctta ttgttacagc tctaagagcc aactcagctg tcaaactaca gaataatgaa 12360
ctgagtccgg tagcactacg acagatgtct tgtgcggctg gtaccacaca aacagcttgt 12420
actgatgaca atgcacttgc ctactataac aattcgaagg gaggtaggtt tgtgctggca 12480
ttactatcag accaccaaga tctcaaatgg gctagattcc ctaagagtga tggtacaggt 12540
acaatttaca cagaactgga accaccttgt aggtttgtta cagacacacc aaaagggcct 12600
aaagtgaaat acttgtactt catcaaaggc ttaaataacc taaatagagg tatggttctg 12660
ggcagtttag ctgctacagt acgtcttcag gctggaaatg ctacagaagt acctgccaac 12720
tcaactgtgc tttccttctg tgcctttgca gtagaccctg ctaaagcata taaggattac 12780
ctagcaagtg gaggacaacc aatcaccaac tgtgtgaaga tgttgtgtac acacactggt 12840
acaggacagg caattactgt aacaccagaa gctaacatgg accaagagtc ctttggtggt 12900
gcttcatgct gtctgtattg tagatgccac attgatcatc caaatcctaa aggattttgt 12960
gacttgaaag gtaagtacgt ccaaatacct accacttgtg ctaatgaccc agtgggtttt 13020
acacttagaa acacagtctg taccgtctgc ggaatgtgga aaggttatgg ctgtagttgt 13080
gatcaactcc gcgaacccat gatgcagtct gcggatgcgt caacgttttt aaacgggttt 13140
gcggtgtaag tgcagcccgt cttacaccgt gcggcacagg cactagcact gatgtcgtct 13200
acagggcttt tgatatttac aacgaaaaag ttgctggttt tgcaaagttc ctaaaaacta 13260
attgctgccg cttccaggag aaggatgagg aaggcaattt attagactct tactttgtag 13320
ttaagaggca tactatgtct aactaccaac atgaagagac tatttataac ttggttaaaa 13380
attgtccagc ggttgctgtt catgattttt tcaagtttag agtagatggt gacatggtac 13440
cacatatatc acgtcagcgt ctaactaaat acacaatggc tgatttagtc tatgctctac 13500
gtcactttga cgagggtaat tgtgacacat taaaagaaat actcgtcaca tacaattgtt 13560
gtgatgatga ttattacaat aagaaggatt ggtatgactt cgtagagaat cctgacatct 13620
tacgcgtata tgctaactta ggtgagcgtg tacgccaagc attattaaag actgtacaat 13680
tctgcgatgc tatgcgtgat gcaggcattg taggcgtact gacattagat aatcaggatc 13740
ttaatgggaa ctggtacgat ttcggtgatt tcgtacaagt agcaccaggc tgcggagttc 13800
ctattgtgga ttcatactac tcattgctga tgcccatcct cactctgact agggcattgg 13860
ctgctgagtc ccatatggat gctgatctcg caaaaccact tattaagtgg gatttgctga 13920
aatatgattt tacggaagag agactttgcc tcttcgaccg ttactttaaa tattgggacc 13980
agacatacca tcccaattgt attaactgtt tggatgatag gtgtatcctt cattgtgcaa 14040
actttaatgt gttattttct actgtgtttc cacctacaag ttttggacca ctagtaagaa 14100
aaatatttgt agatggtgtt ccttttgttg tttcaactgg ataccatttt cgtgagttag 14160
gagttgtaca taatcaggat gtaaacttac atagctcgcg tctcagtttc aaggaacttt 14220
tagtgtatgc tgctgatcca gccatgcatg cagcttctgg caatttattg ctagataaac 14280
gcactacatg cttttcggta gctgcactaa caaacaatgt tgcttttcaa actgtcaaac 14340
ccggtaattt taacaaagac ttttatgact ttgctgtgtc taaaggtttc tttaaggaag 14400
gaagttctgt tgaactaaaa cacttcttct ttgctcagga tggcaatgct gctatcagtg 14460
attatgacta ttatcgttat aatctgccaa caatgtgtga tatcagacaa ctcctattcg 14520
tagttgaagt tgttgataaa tactttgatt gttacgatgg tggctgtatt aatgccaacc 14580
aagtaatcgt taacaatctg gacaaatcag ctggtttccc atttaataaa tggggtaagg 14640
ctagacttta ttatgactca atgagttatg aggatcaaga tgcacttttc gcgtatacta 14700
agcgtaatgt catccctact ataactcaaa tgaatcttaa gtatgccatt agtgcaaaga 14760
atagagctcg cactgtagct ggtgtctcta tctgtagtac tatgacaaat agacagtttc 14820
atcagaaatt attgaagtca atagccgcca ctagaggagc tactgtggta attggaacaa 14880
gcaaatttta cggtggctgg cataatatgt taaaaactgt ttacagtgat gtagaaactc 14940
cacaccttat gggttgggat tatccaaaat gtgacagagc catgcctaac atgcttagga 15000
taatggcctc tcttgttctt gctcgcaaac atagcacttg ctgtaactta tcacaccgtt 15060
tctacaggtt agctaacgag tgtgcgcaag tattaagtga gatggtcatg tgtggcggct 15120
cactatatgt taaaccaggt ggaacatcat ccggtgatgc tacaactgct tatgctaata 15180
gtgtctttaa catttgtcaa gctgttacag ccaacgtaaa tgcacttctt tcaactgatg 15240
gtaataagat agctgacaag tatgtccgca atctacaaca caggctctat gagtgtctct 15300
atagaaatag ggatgttgat cacgaattcg tggatgagtt ttacgcttac ctgcgtaaac 15360
atttctccat gatgattctt tctgatgacg ccgttgtgtg ctataacagt aactatgcgg 15420
ctcaaggttt agtagctagc attaagaact ttaaggcagt tctttattat caaaataatg 15480
tgttcatgtc tgaggcaaaa tgttggactg agactgacct tactaaagga cctcacgaat 15540
tttgctcaca gcatacaatg ctagttaaac aaggagatga ctacgtgtac ctgccttacc 15600
cagatccatc aagaatatta ggcgcgggct gttttgtcga tgatattgtc aaaacagatg 15660
gtacacttat gattgagagg tttgtgtcat tagctattga tgcctacccc cttactaaac 15720
atcctaatca ggagtatgct gatgtctttc acttgtattt acaatacatt aggaagttac 15780
atgatgagct tactggtcac atgctagaca tgtattctgt aatgctaacc aatgacaaca 15840
cctcacggta ttgggaacct gagttttatg aagctatgta cacaccacac acagtcttgc 15900
aggctgtagg tgcttgtgta ttgtgtaatt cacagacttc acttcgttgc ggtgcctgca 15960
ttaggagacc attcctgtgc tgcaagtgct gctatgacca tgtcatttca acatcacata 16020
aattagtgtt gtctgttaat ccctatgttt gcaatgcacc aggttgtgat gtcactgacg 16080
tgacacaact atatctagga ggtatgagct attactgcaa gtcacataag cctcccatta 16140
gttttccatt gtgtgctaat ggtcaggttt ttggtttata caagaacaca tgtgtaggta 16200
gtgacaatgt cactgacttc aatgcgatag caacatgtga ttggactaat gctggcgatt 16260
acatacttgc caacacttgt actgagagac tcaaactctt tgcagcagaa acactcaaag 16320
ctactgagga aacattcaag ctgtcatatg gtattgccac tgtacgtgaa gtactctctg 16380
acagagaatt gcatctttca tgggaggttg gaaaacctag accaccattg aatagaaatt 16440
atgtctttac tggttaccgt gtaactaaaa atagtaaagt acagattgga gagtacacct 16500
ttgaaaaagg tgactatggt gatgctgttg tgtacagagg tactacgaca tacaaattga 16560
atgttggtga ttactttgtg ttgacatctc acactgtaat gccacttagt gcacctactc 16620
tagtgccaca agagcactat gtgagaatta ctggcttgta cccaacactc aacatctcag 16680
atgagttttc tagcaatgtt gcaaattatc aaaaggtcgg tatgcaaaag tactctacac 16740
tccagggccc acctggcact ggtaagagtc attttgccat cggacttgct ctctactacc 16800
catctgctcg catagtgtat acagcttgct cccatgcagc tgttgatgcc ctatgcgaaa 16860
aggcactaaa atacttgccc atagataaat gtagtagaat catacctgcg cgtgctcgcg 16920
tagagtgttt tgacaaattc aaagtgaatt caacactaga acagtatgtt ttctgcactg 16980
taaatgcatt gccagaaaca actgctgata ttgtagtctt tgatgaaatc tctatggcta 17040
ctaattatga cttgagtgtt gtcaatgcta gacttcgtgc aaaacactac gtctatattg 17100
gcgatcctgc tcaattacca gccccgcgca cattgctgac caaaggcaca ctagaaccag 17160
aatacttcaa ttcagtgtgc agacttatga aaacaatagg tccagacatg ttccttggaa 17220
cttgtcgccg ttgtcctgct gaaattgtcg acactgtgag tgctttagtt tatgataata 17280
agctaaaggc acacaaggag aagtcagctc aatgtttcaa aatgttttat aagggtgtta 17340
ttacacatga tgtttcatct gcaattaaca gacctcaaat aggcgttgta agagaatttc 17400
ttacacgcaa tcctgcttgg agaaaagctg tttttatctc accttataat tcacagaatg 17460
ctgtagcttc aaaaatctta ggattgccta cgcagactgt cgattcctcc cagggttctg 17520
agtatgacta tgtcatattc acacaaacta ctgaaacagc acactcttgc aatgttaacc 17580
gctttaatgt ggctatcaca agagcaaaaa ttggcatttt gtgcataatg tccgatagag 17640
atctttatga caaactgcaa ttcacaagtt tagaagtacc acgccgcaat gtggctacat 17700
tacaggcaga aaatgtaact ggacttttta aggactgtag taagatcatc accggtcttc 17760
atccaacaca ggcacctaca cacctcagcg ttgatacaaa atttaagact gagggactat 17820
gtgttgacat accaggcata ccaaaggaca tgacctaccg tagactcatc tctatgatgg 17880
gtttcaaaat gaattaccaa gttaatggtt accctaatat gtttattacc cgtgaggaag 17940
ctattcgtca cgttcgtgca tggattggct tcgacgtaga gggctgtcat gcaactagag 18000
atgctgtggg tactaaccta cctctccagc taggattttc tacaggtgtt aacttagtag 18060
ctgtaccgac tggctatgtt gacactgaaa ataacacaga attcaccaga gttaatgcaa 18120
gacctccacc aggtgaccag tttaaacatc ttataccact catgtacaaa ggcttgccct 18180
ggaatgtagt gcgtattaag atagtacaaa tgctcagtga tacactgaaa ggattgtcag 18240
acagagtcgt gtttgtcctt tgggcgcatg gctttgagct tacatcaatg aagtactttg 18300
tcaagattgg acctgaaaga acgtgctgtc tgtgtgacaa acgtgcaact tgcttttcta 18360
cttcatcaga tacttatgcc tgctggaatc attctgtggg ttttgactat gtctacaacc 18420
catttatgat tgatgtccag cagtggggtt ttacgggtaa ccttcagagt aaccacgacc 18480
aacattgtca agtgcatgga aatgcacatg tggctagttg tgatgctatc atgactagat 18540
gcttggcggt ccatgagtgc tttgttaagc gcgttgattg gtctgttgaa taccccatta 18600
taggagatga actgaagatt aattccgctt gcagaaaagt acagcatatg gttgtaaagt 18660
ctgcattgct tgctgataag tttccagttc ttcatgacat tggaaatcca aaggctatta 18720
agtgtgtgcc tcaggcggaa gtagaatgga agttctatga tgctcagcca tgcagtgaca 18780
aagcctataa aatagaggaa cttttctatt cctacgctac acatcatgat aaattcactg 18840
atggtgtttg tttgttttgg aactgtaacg ttgatcgtta cccagccaat gcaattgtat 18900
gtaggtttga cacgagagtt ttgtcaaact tgaatttacc aggttgtgac ggtggtagtt 18960
tgtatgtgaa taagcatgca ttccacactc cagcttttga taaaagtgca tttaccaatt 19020
taaagcaatt gcctttcttt tattattctg acagtccttg tgagtctcat ggcaaacaag 19080
tagtgtcaga tattgattat gtaccactta aatctgctac gtgtattaca cgatgcaatt 19140
tgggaggtgc tgtttgcaga caccatgcaa atgagtaccg acagtactta gatgcatata 19200
acatgatgat ttctgctgga tttagcctat ggatttacaa acagtttgat acttataatc 19260
tgtggaatac attcaccagg ttacagagtt tagaaaatgt ggcttacaat gttgttaaca 19320
aaggacactt cgatggacaa gctggtgaag cacctgtttc catcattaat aatgctgttt 19380
acacaaaggt agatggtgtt gatgtagaga tctttgaaaa caagacaaca cttcctgtta 19440
atgttgcgtt tgagctttgg gctaagcgta acattaaacc agtgccagag attaagatac 19500
tcaataactt gggtgtcgat atcgctgcta atactgtggt ctgggactac aagagagaag 19560
caccagcaca tatgtcaaca ataggtgtct gtacaatgac cgacattgct aagaaaccta 19620
ctgagagtgc ttgttcctcg cttactgtct tatttgatgg tagagtggaa ggacaggtag 19680
acctttttag aaatgcccgt aatggtgttt taataacaga aggttcagtt aaaggtttaa 19740
caccttcaaa ggggccagca caagctagtg tcaatggagt cacattaatt ggagaatccg 19800
taaaaacaca gtttaactat tttaagaaag tagatggcat tattcaacag ttgcctgaaa 19860
cctactttac tcagagccga gacttagagg atttcaagcc cagatcacaa atggaaactg 19920
actttcttga gctcgctatg gatgaattca tacaacggta taagctagag ggttatgcct 19980
tcgaacatat cgtttatggg gatttcagtc atggacaact tggcggcctt catctaatga 20040
ttggtttagc caagcgctca caagattcac cgcttaaatt agaggacttt atccctatgg 20100
atagcacagt gaaaaattac ttcataacag atgcacaaac aggttcatca aaatgtgtgt 20160
gctctgttat tgacctttta cttgatgact ttgttgagat aataaagtca caggatttgt 20220
cagtaatttc taaggtagtc aaggttacaa ttgactacgc tgagatttca ttcatgcttt 20280
ggtgtaaaga tggacacgtt gaaaccttct accctaaatt acaagcaagt caagcgtggc 20340
aaccgggggt tgcaatgcct aacttgtaca agatgcaaag aatgcttctt gaaaagtgtg 20400
accttcagaa ttatggtgaa aatgctgtca taccaaaagg aataatgatg aatgtcgcaa 20460
aatacactca actgtgtcaa tacttaaata cacttacttt agctgtaccc tacaacatga 20520
gagttattca ctttggtgct ggatctgata aaggagttgc accaggtaca gctgtactca 20580
gacaatggtt gccaactggc acactacttg tcgattcaga ccttaatgac ttcgtctctg 20640
acgcggattc tactttaatt ggagactgtg caacagtaca tacggctaat aaatgggatc 20700
tcattattag cgatatgtat gaccctaaga caaaacatgt gacaaaagag aacgactcaa 20760
aagaagggtt tttcacttac ctgtgtggat ttataaagca aaagctagcc ttgggtggct 20820
ctgtggctgt gaagataaca gagcattctt ggaatgctga tctttacaag cttatgggac 20880
acttctcatg gtggacagct tttgttacaa atgtaaatgc atcatcatca gaagcatttc 20940
taattggggt taactatcta ggcaagccaa aggaacaaat tgatggctat accatgcatg 21000
ctaactacat cttttggagg aacacaaatc ctattcaatt gtcttcctat tcactttttg 21060
acatgagcaa gtttcctctc aaattaagag ggactgctgt tatgtattta aaggagaatc 21120
agatcaatga catgatttat tctctacttg agaaaggtag acttatcatt agagagagta 21180
acaaagttgt ggtgtctagt gatattttag ttaataacta aacgaacatg aaattgttag 21240
ttttagtttt tgctactcta gtctcctctt acactataga gaagtgcctt gattttgatg 21300
accgcactcc acctgcaaat actcaatttt tatcttctca cagaggtgtt tattacccag 21360
atgacatttt taggtctaat gtcttgcatt tagtacaaga tcatttccta ccttttgact 21420
ctaacgtcac caggtttata acgtttggcc taaattttga taatcccata atacccttca 21480
gggatggtat ttattttgct gcgactgaaa agtctaatgt tattagagga tgggtttttg 21540
gttctacaat gaacaacaaa tctcaatccg ttataataat gaacaactca actaatttag 21600
tcattagggc ttgtaatttt gagttgtgtg acaatccatt ttttgttgtg ttgaaatcta 21660
acaacactca aataccatct tacatattta ataatgcatt caattgcaca tttgaatatg 21720
tttctaagga ttttaaccta gaccttggtg aaaaaccagg taatttcaag gatctcagag 21780
agtttgtttt caggaataaa gatggttttt tgcatgttta ttccggttac caacccattt 21840
ctgctgccag tggtttgcca actggtttta atgcactcaa acctattttc aagttacctc 21900
tgggtattaa tattactaat ttcagaacac ttctgactgc ttttccgcct agacctgatt 21960
attggggtac ttcagctgca gcttattttg taggatattt aaaaccaact acattcatgc 22020
tcaagtatga tgaaaatggt acaatcacag atgctgtcga ttgttctcaa aatccacttg 22080
ctgaactcaa atgctctgtt aaaagttttg agattgacaa aggaatttat caaacctcca 22140
attttagggt agcaccctca aaggaagttg tgaggttccc taatattaca aacctgtgtc 22200
cttttgggga ggtttttaat gctactacat ttccttctgt ctatgcatgg gagaggaaaa 22260
gaatttctaa ttgtgttgct gattactctg tactctacaa ctcaacatct ttttcaactt 22320
ttaagtgtta tggcgtttct gccactaagc tgaatgacct ttgcttctcc aacgtctatg 22380
cagattcatt cgtagtcaaa ggagatgatg taaggcaaat agcaccagga cagaccggtg 22440
ttattgctga ttataattac aaattaccag atgacttctt gggttgtgtc ctagcatgga 22500
acaccaattc taaagattct tccacttccg gtaattataa ttatttatat agatgggtta 22560
gaaggtctaa gcttaaccct tatgagcgcg acttatctaa cgacatctat tcacctggag 22620
gtcagtcttg ctcagctgta ggtcctaatt gttataaccc cttacgtcca tatggctttt 22680
ttacaacagc tggtgttgga caccaacctt atagagttgt agtactttct tttgaacttt 22740
taaatgcacc cgctacagtc tgtggaccaa aattatccac cgaccttatt aaaaatcaat 22800
gtgtcaattt taactttaat ggactcactg gtactggtgt gttaactcct tcttcaaaga 22860
gatttcaacc atttcaacaa tttggtcgtg atgtttcgga tttcactgat tcagttcgag 22920
atccgaagac gtctgaaata ttagacattt caccttgctc ttttggcggt gtaagtgtaa 22980
tcacacctgg aacaaatact tcatcagaag ttgctgttct atatcaagat gttaactgca 23040
ctgatgttcc tgtagcaatc catgcagacc aactcacacc ttcttggcgc gtatactcta 23100
ctggaaataa tgtatttcaa acccaggcag gctgtcttat aggagctgag catgtcgaca 23160
cttcttatga gtgcgacatt cctattggag ctggcatttg tgctagttac catacagttt 23220
cttcattacg tagtactagc caaaaatcta ttgtggctta tactatgtct ttaggtgctg 23280
atagttcaat tgcttactct aataacacca ttgctatacc tactaacttt tcaattagca 23340
ttactacaga agtaatgcct gtttctatgg ctaaaacctc tgtagattgt aatatgtaca 23400
tctgcggaga ttctactgaa tgtgctaatt tgcttctcca atatggtagc ttttgcacac 23460
aactaaatcg tgcactctca ggtattgctg ttgaacagga tcgcaacaca cgtgaagtgt 23520
tcgctcaagt caaacaaatg tacaaaaccc caactttgaa agattttggt ggttttaatt 23580
tttcacaaat attacctgac cctctaaagc caactaagag gtcttttatt gaggacttgc 23640
tctttaataa ggtgacactc gctgatgctg gctttatgaa gcaatatggc gaatgcctag 23700
gtgatattaa tgctagagat ctcatttgtg cgcagaagtt caatggactt acagtgctgc 23760
cacctctgct cactgatgat atgattgctg cctacactgc tgctctagtt agtggtactg 23820
ccactgctgg atggacattc ggtgctggcg ctgctcttca aatacctttt gctatgcaaa 23880
tggcatatag gttcaatggc attggagtta ctcaaaatgt tctctatgag aaccaaaaac 23940
aaatcgccaa tcaatttaac aaggcgatca gccaaattca agaatcactc acaacaacat 24000
ccactgcatt gggcaagctg caagatgtcg tcaaccagaa tgctcaagca ttaaacacac 24060
ttgttaaaca acttagctcc aattttggtg cgatttcaag tgtgctaaat gatatccttt 24120
cgcgacttga taaagtcgag gcagaggtac aaattgacag gttaattaca ggcagactgc 24180
aaagccttca aacctatgta acacaacaac taatcagggc tgctgaaatc agggcttctg 24240
ctaatcttgc tgctactaaa atgtctgagt gtgttcttgg acaatcaaaa agagttgact 24300
tttgcggaaa gggctaccat cttatgtcct tcccacaagc agccccgcat ggtgttgtct 24360
tcctacatgt cacatatgtg ccatctcaag agagaaactt caccacagcg ccagcaattt 24420
gtcatgaagg caaagcatac ttccctcgtg aaggtgtttt tgtgtttaat ggcacttcgt 24480
ggtttattac acagaggaac ttcttttctc cacaaataat tactacagac aatacatttg 24540
tctccggaag ttgtgatgtc gtaattggca tcattaacaa cacagtttat gatcctctgc 24600
aacctgagct tgactcattc aaagaagagc tggacaagta cttcaaaaat cacacatcac 24660
cagatgttga tcttggcgac atttcaggca ttaacgcttc tgtcgtcaac attcaaaaag 24720
aaattgaccg cctcaatgag gtcgctaaaa atttaaatga atcactcatt gaccttcaag 24780
aattgggaaa atatgagcaa tatattaaat ggccttggta tgtttggctc ggcttcattg 24840
ctggactaat tgccatcgtc atggttacaa tcttgctttg ttgcatgact agttgttgca 24900
gttgcctcaa gggtgcatgc tcttgtggtt cttgctgcaa gtttgatgag gatgactctg 24960
agccagttct caagggtgtc aaattacatt acacataaac gaacttatgg atttgtttat 25020
gagaattttt actcttggac caattactgc acagccagga aaaattgaca atgcttctcc 25080
tgcaagtact gttcatgcta cagcaacgat accgctacaa gcctcactcc ctttcggatg 25140
gcttgttatt ggcgttgcat ttcttgctgt ttttcagagc gctaccaaaa taatttcgct 25200
caataaaaga tggcagctag ccctttataa gggcttccag ttcatttgca atttactgct 25260
gctatttgtt accatctatt cacatctttt gcttgtcgct gcaggtatgg aggcgcaatt 25320
tttgtacctc tatgccttga tatattttct acaatgcatc aacgcatgta gaattatcat 25380
gagatgttgg ctttgttgga agtgcaaatc caagaaccca ttactttatg atgccaacta 25440
ctttgtttgc tggcacacac ataactatga ctactgtata ccgtataaca gtgtcacaga 25500
tacaattgtc gttactgcag gtgacggcat ttcaacacca aaactcaaag aagactacca 25560
aattggtggt tattctgaga attggcactc aggtgttaaa gactatgtcg ttgtacatgg 25620
ctatttcacc gaagtttact accagcttga gtctacacaa attactacag acactggtat 25680
tgaaaatgct acattcttca tctttaacaa gcttgttaaa gacccaccga atgtgcaaat 25740
acacacaatc gacggctctt caggagttgt aaatccagca atggatccaa tttatgatga 25800
gccgacgacg actactagcg tgcctttgta agcacaagaa agtgagtacg aacttatgta 25860
ctcattcgtt tcggaagaaa caggtacgtt aatagttaat agcgtacttc tttttcttgc 25920
tttcgtggta ttcttgctag tcacactagc catccttact gcgcttcgat tgtgtgcgta 25980
ctgctgcaat attgttaacg tgagtttagt aaaaccaact gtttacgttt actcgcgtgt 26040
taaaaatctg aactcttctg aaggagttcc tgatcttctg gtctaaacga actaactatt 26100
attattattc tgtttggaac tttaacattg cttatcatgg ctgagaacgg gactatttcc 26160
gttgaggagc ttaaaagact cctggaacaa tggaacctag taataggttt cctattccta 26220
gcctggatta tgttactaca atttgcctat tctaatcgga acaggttttt gtacataata 26280
aagcttgttt tcctgtggct cttgtggcca gtaacacttg cttgctttgt gcttgctgct 26340
gtttacagaa ttaattgggt gactggcggg attgcgattg caatggcttg tattgtaggc 26400
ttgatgtggc ttagctactt cattgcttcc ttcaggctat ttgctcgtac ccgctcaatg 26460
tggtcattca acccagaaac aaacattctt ctcaatgtgc ctcttcgagg gacaattgtg 26520
accagaccgc tcatggaaag tgaacttgtc attggcgctg tgatcattcg tggtcacttg 26580
cgcatggctg gacactccct agggcgctgt gacatcaagg acctgccaaa agagatcact 26640
gtggctacat cacgaacgct ttcttattac aaattaggag cgtcgcagcg tgtaggcact 26700
gattcaggtt ttgctgcata caaccgctac cgtattggaa actacaaatt aaatacagac 26760
cacgccggta gcaacgacaa tattgctttg ctagtacagt aagtgacaac agatgtttca 26820
tcttgttgac ttccaggtta caatagcaga gatattgatt atcattatga ggactttcag 26880
gattgctatt tggaatcttg acatgataat aagttcaata gtgagacaat tatttaagcc 26940
tctaactaag aataaatatt cagagttaga tgatgaagaa cctatggaga tagattatcc 27000
ttgataaacg aaccactatg ttacttttag taacattgtt tggtttagca tcagggtgca 27060
gcttaccact tacggttagc tgccctagag gcctaccttt cactctacag attaacacta 27120
ctagtgttac tgtggagtgg tatcgggtat ctcctgcatc aatgcaaggt cttacaaaga 27180
taaatactgg cagcactatt tttgataaca actttagtgt agtcaataat aatttgtact 27240
tcaaacagtg ttttggaggc ttttttttac agcacgctgt taccgccagg gtaagcatga 27300
cggtgctata g 27311
<210> 2
<211> 27425
<212> DNA
<213> WIV1-RsSHC014S mouse adapted strain (Artificial Sequence)
<400> 2
atggagagcc ttgttcttgg tgtcaacgag aaaacacacg tccaactcag tttgcctgtt 60
cttcaggtta gagacgtgct agtgcgtggc ttcggggact ctgtggaaga ggccctatcg 120
gaggcacgtg aacaccttaa aaatggcact tgtggtttag tagagctgga aaaaggcgta 180
ctgccccagc ttgaacagcc ctatgtgttc attaaacgtt ctgatgcctt aagcaccaat 240
cacggccaca aggtcgttga gttggttgca gaattggacg gcattcagta cggtcgtagc 300
ggtataactc tgggagtgct cgtgccacat gtgggcgaaa ccccaatcgc ataccgcaat 360
gttcttcttc gtaagaacgg taataaggga gccggtggcc atagctttgg catcgatcta 420
aagtcttatg acttaggtga cgagcttggc actgatccca ttgaagatta tgaacaaaac 480
tggaacacta agcatggcag tggtgtactc cgtgaactca ctcgcgagct caatggaggt 540
gcattcactc gctatgtcga taacaacttt tgtggcccag atgggtaccc tcttgattgc 600
attaaagatt ttctcgctcg cgcgggcaag tcaatgtgca ctctttctga acaacttgat 660
tacatcgagt cgaagagagg tgtctactgc tgccgtgacc atgagcatga agttgcttgg 720
ttcactgagc gctctgataa gagctatgag catcagacac ccttcgaaat taagagtgcc 780
aagaaatttg acaccttcaa aggggaatgc ccaaagtttg tatttcctct cgattcaaaa 840
gtcaaagtca ttcaaccacg tgttgaaaag aaaaagactg aaggtttcat ggggcgcata 900
cgctctgtgt accctgttgc atctccacag gagtgtaaca acatgcactt gtctaccttg 960
atgaaatgta atcattgcga tgaagtttca tggcagacgt gcgattttct gaaagccact 1020
tgtgaacatt gtggcactga aaactcagtc acagaaggac ctactacatg tgggtaccta 1080
cctactaatg ctgtagtgaa aatgccatgt cccgcctgtc aagacccgga gattggacct 1140
gagcatagtg tcgcagacta tcacaaccac tcaaacattg aaactcgact ccgcaaagga 1200
ggtaggacta gatgttttgg aggctgtgtg tttgcctatg tcggctgcta caacaagcgt 1260
gcctactggg ttcctcgtgc tagtgctgat attggttcag gccatactgg aattactggt 1320
gacaacgtgg agaccttgaa tgaggatctc cttgagatac tgagtcgtga acgtgttaat 1380
attaacattg ttggcgattt tcagttgaat gaagaggtgg ccatcatttt ggcatctttt 1440
tctgcttcta caagtgcctt tattgacact ataaggagtc ttgattacaa gtctttcaaa 1500
gccattgttg agtcctgcgg taactacaaa gttaccaagg gaaagcccgt aaaaggtgct 1560
tggaacattg gacaacagag atcagtttta acaccactgt gtggtttccc ctcacaggct 1620
gctggtgtta tcagatcaat ttttgcacgc acacttgatg cagcaaacca ctcaattcct 1680
gatttgcaaa gagcagctgt caccatactt gatggtattt ctgaacagtc attacgtctt 1740
gtcgacgtca tggtttatac ctcagacctg cttaccaaca gtgtcatcat tatggcatat 1800
gtaactggtg gtcttgtaca acagacttct cagtggttgt ctaatttgtt gggcactact 1860
gttgaaaaac tcaggcccat ctttgtgtgg attgaggcga aacttagtgc aggagttgaa 1920
tttctcaagg atgcttggga gattctcaaa tttctaatta caggtgtttt tgacatcgtc 1980
aagggtcaaa tacaggttgc ttcagataac atcaagggtt gtgtaaaatg cttcattgat 2040
gttgttaata aagcactcga aatgtgcatt gaccaagtca ctatcgctgg cactaagttg 2100
cgatcactca acttaggtga agtcttcatc gctcaaagca agggacttta ccgtcagtgt 2160
atacgtggca aggaacagct gcaactactt atgcctctta aggcaccaaa agaagtcacc 2220
tttcttgaag gtgattcaca tgacacagta cttacctctg aggaggttgt tctcaaaaac 2280
ggtgaactcg aggcactcga gacgcctgtt gatagcttca caaatggagc tgtagttggc 2340
acaccagttt gtgtaaatgg cctcatgctc ttagagatca aggacaaaga acaatactgc 2400
gcattgtctc ctggtttact ggctacaaac aatgtctttc gcctaaaagg gggtgcacca 2460
actaaaggtg taacctttgg agaagatact gttttggaag ttcaaggtta caagaatgtg 2520
agaatcacat ttgagcttga tgaacgtgtt gacaaagtgc ttaatgaaaa gtgctctgtc 2580
tacactgttg aatccggtac cgaagttact gagtttgcat gtgttgtagc agaggctgtt 2640
gtgaagactt tacaaccagt ttctgatctt cttaccaaca tgggtattga tcttgatgaa 2700
tggagtgtgg ctacattcta tttgtttgat gatgctggtg aagaaaaact ttcttcacgt 2760
atgtactgtt ccttttatcc tcctgatgat gaggaggatt gtgatgagta tgatgaagaa 2820
gaggaagtcc tggaagaatc ctgtgcgcat gaatacggta cagaagaaga ttaccaaggt 2880
ctttcactgg aatttggtgc ctcaactgaa atgcaagttg aagaagaaga agaagaggac 2940
tggcttggtg atgctactga attatcggag catgaaccag aaccagaact aacacctgaa 3000
gaaccagtta accagtttac tggttattta aaacttactg acaatgttgc cattaagtgt 3060
gtggacatcg tgaaggaggc gcaaaacgct aaccccacgg tgattgtaaa tgctgctaac 3120
atacatctga aacatggtgg tggtgtagca ggtgcactca acaaggcaac caacggtgcc 3180
atgcaaaaag agagcgatga ttacattaag ctaaatggtc ctctcgtagt tggaggttca 3240
tgtttgcttt ctggacataa tcttgctaag aagtgtctgc atgttgttgg acctaaccta 3300
aatgcaggtg aggacatcca gcttcttaag gcagcatatg aaaattttaa ttcacaggac 3360
actttacttg caccattgtt gtcagcaggc atatttggtg ctaaaccact tcagtcttta 3420
caagtgtgcg tgcagacagt tcgtacacag gtttacattg cagtcaatga caaagctctt 3480
tatgagcagg ttgtcatgga ttaccttgat agcctgaagc ctagagtgga agcacctaaa 3540
caagaggagc caccaaggac agaagatcct aaaattgagg agaaatctgt cgtacagaag 3600
cctatcgatg tgaagccaaa aattaaggct tgcattgatg aggttaccac aacactggaa 3660
gaaactaagt ttcttaccaa taagttactc ttgtttgctg acatcaatgg taagctttac 3720
catgattctc acaacatgct tagaggtgaa gatatgtctt tccttgagaa ggatgcacct 3780
tacgtggtag gtgatgttat cactagtggt gatatcactt gtgttgtaat accctccaaa 3840
aaggctggtg gcactacaga gatgctctca agagctttga agaaagtgcc agttgatgag 3900
tatataacca cataccctgg acaaggatgt gctggttata cacttgagga agctaagact 3960
gctcttaaga aatgcaaatc tgcattttac gtgttacctt cagaaacacc taatgctaag 4020
gaagagattc taggaactgt gtcctggaat ttgagagaaa tgcttgctca tgctgaagag 4080
acaagaaaat taatgcctat atgcatggat gttagagcca taatggccac catccaacgc 4140
aagtacaaag gaattaaagt tcaagaaggc attgttgact atggagtccg attcttcttt 4200
tatactagta aagagcctgt agcttctatc attacgaagc tgaactctct aaatgagcca 4260
cttgtcacaa tgccaattgg ttatgtgaca catggtttta atcttgaaga ggctgcgcgc 4320
tgtatgcgtt ctcttaaagc tcctgccgta gtgtcagtat catcaccaga tgctgttact 4380
acatataatg gatacctcac ttcgtcatca aagacatctg aggagcactt tgtggaaaca 4440
gtttctttgg ctggttctta cagagattgg tactattcag gacagcgtac agagttaggt 4500
gttgaatttc ttaagcgtgg tgacaaaatt gtgtaccaca ctttggagag tcccgtcgag 4560
tttcatcttg acggtgaggt tcttccactt gacaaactaa agagtctttt atccctacgg 4620
gaggttaaga ctataaaagt gttcacaact gtggacaaca ctaatctcca cacacagctt 4680
gtggatatgt ctatgacata tggacagcag tttggtccaa catacttgga tggtgctgat 4740
gttacaaaaa ttaaacctca tgtaaatcat gagggtaaga ctttctttgt actacctagt 4800
gatgacacac tacgtagtga agctttcgag tactaccaca ctcttgatga gagttttctt 4860
ggtaggtaca tgtctgcttt aaaccacaca aagaaatgga aatttcctca agttggtggt 4920
ttaacttcaa ttaaatgggc tgataacaat tgttatttgt ctagtgtttt attagcactt 4980
caacagattg aagtcaaatt caatgcacca gcacttcaag aggcttatta tagagcccgt 5040
gctggtgatg ctgctaactt ttgtgcactc atacttgctt acagtaataa aactgttggc 5100
gagctcggtg atgtcagaga aactatgacc catcttctac agcatgctaa tttggaatcc 5160
gcaaagcgag ttcttaatgt ggtgtgtaaa cattgcggtc agaaaactac caccttaacg 5220
ggtgtagaag ccgtgatgta catgggtact ctatcttatg ataatcttaa gacaggtgtt 5280
tccattccat gtgtgtgtgg tcgcgatgct acacaatatc tagtacaaca agagtcttct 5340
tttgttatga tgtctgcacc acctgctgaa tataaattac agcaaggtac atttttatgt 5400
gcgaatgagt acactggtaa ctatcagtgt ggtcattaca ctcatataac tgctaaggag 5460
accctctatc gtattgatgg agctcacctt acaaagatgt cagagtacaa aggaccagtg 5520
actgacgttt tctataagga aacatcttac actacaacca tcaagcctgt gtcgtataaa 5580
ctcgatggag ttacttacac agagattgaa ccaaaattgg atgggtatta taaaaaggat 5640
aatgcttact atacagagca gcctatagac cttgtaccaa ctcaaccact accaaatgcg 5700
agttttgaca atttcaaact cacatgttct aatacaaaat ttgctgatga tttaaatcaa 5760
atgacaggct tcacaaagcc agcttcacga gagttatatg tcacattctt tccagacttg 5820
aatggcgatg tagtggctat tgactataga cactattcag cgagtttcaa gaaaggtgct 5880
aaattactgc ataagccaat tgtttggcat atcaatcagg ctacaaccaa gacaacgttt 5940
aaaccaaaca cttggtgttt acgttgtctt tggagtacaa agccagtaga tacttcaaat 6000
tcatttgaag ttctggcagt agaagacaca caaggaatgg acaatcttgc ttgtgaaagt 6060
caacaaccca cctctgaaga agtagtggaa aatcctacca tacagaagga agtcatagag 6120
tgtgacgtga aaactactga agttgtaggc aatgtcatac ttaaaccatc agatgaaggt 6180
gttaaagtaa cacaagagtt ggatcatgag gatcttatgg ctgcttatgt ggaaaataca 6240
agcattacca ttaagaaacc taatgagctt tcactagcct taggtttaaa aacaattgcc 6300
actcatggta ttgctgcaat taatagtgtg ccttggagta aaattttggc ttatgtcaaa 6360
ccattcttag gacaagcagc aattacaaca tcaaattgcg ctaagagatt agcacaacgt 6420
gtgtttaaca attatatgcc ttatgtgctt acactattgt ttcaattgtg tacttttaca 6480
aaaagtacaa attctagaat tagagcttca ctacctacga ctattgctaa aaatagtgtt 6540
aggggtgttg ctagattatg tttggatgct ggcattaatt atgtaaagtc acccaaattt 6600
tctaaattgt tcactattgc aatgtggcta ttattgttaa gcatttgctt aggttcacta 6660
atctatgtaa ctgcagcttt aggtgtatta ttgtccaact ttggagctcc ttcttattgt 6720
agtggcgtta gagaatcgta cctcaattcc tctaatgtta ctactatgga tttctgtgaa 6780
ggttcttttc cttgcagtgt ttgtttaagt ggattagact cgcttgattc ctatccagct 6840
cttgaaacca tccaggtgac gatctcatcg tacaagctag acttgacaat tttaggtctg 6900
gctgctgagt ggtttttggc atatatgttg tttacaaaat tcttttattt actaggtctt 6960
tcagctataa tgcaggtgtt ctttggctat tttgctagtc atttcatcag caattcttgg 7020
ctcatgtggt ttatcattag cattgtacaa atggcacccg tttctgcaat ggttaggatg 7080
tacatcttct ttgcttcttt ttactacata tggaagagct atgttcatat tatggatggt 7140
tgtacctctt cgacttgcat gatgtgctat aagcgcaatc gtgccacacg cgttgagtgt 7200
acaactattg ttaatggcat gaagagatct ttctatgtct atgcaaatgg aggccgtggc 7260
ttctgcaaga ctcacaattg gaattgtctc aattgtgaca cattttgcac tggtagtaca 7320
ttcattagtg atgaagttgc tcgtgatttg tcactccagt ttaaaagacc aattaaccct 7380
actgaccagt catcgtatat tgttgatagt gttgctgtta aaaatggcgc gcttcatctc 7440
tactttgaca aggctggtca aaagacttat gagagacacc cactctccca ttttatcaat 7500
ttagacaatt tgagagctaa caacactaaa ggttcactac ctattaatgt catagtcttt 7560
gatggcaagt ccaaatgcga cgagtctgct gctaggtctg catctgtgta ctacagtcag 7620
ctaatgtgcc aacctattct gttgcttgac caagctctca tatcagatgt tggagatagt 7680
actgaagttt ctgttaagat gtttgatgct tatgtcgaca ccttttcagc aacttttagt 7740
gtccctatgg aaaaacttag ggccctcgtg gctacagctc atagcgagct ggcaaagggt 7800
gtagctttag atggtgtcct ttctacattt gtgtcagctg ctcgtcaagg tgttgttgat 7860
actgatgttg acacaaagga tgtcattgaa tgtctcaaac tttcacatca ctccgacttg 7920
gaagtgacag gtgacagttg taacaacttc atgctcacct ataacaaagt tgaaaacatg 7980
acgcctagag atcttggcgc atgtattgat tgtaatgcaa ggcatattaa tgctcaagta 8040
gcaaaaagtc acaatgtttc actcatctgg aatgtaaaag actatatgtc tttatctgaa 8100
cagctgcgta aacaaattcg tagtgctgct aagaagaaca acataccttt tagactaact 8160
tgtgctacaa ctagacaggt tgtcaatgtc ataactacta aaatctcact caagggtggt 8220
aagattgtta gtacttggtt taaacttatg cttaaggcca cattattgtg cgtcattgct 8280
acattggtct gttacatcgt tatgccagta catacattgt ctgttcatga tggttacaca 8340
aatgaaatca ttggttacaa agccattcag gatggtgtca ctcgtgacat cgtttctact 8400
gatgattgtt ttgcaaacaa acatgctggt tttgactcat ggcttagcca gcgtggtggt 8460
tcatacaaaa atgacaaaag ctgccctgta gtagctgcta tcattacaag agagattggt 8520
ttcatagtgc ctggcttacc aggtactgtg ttgagagcaa tcaatggtga cttcttgcat 8580
tttctacctc gtgtctttag tgctgttggc aacatttgct acacaccttc taaactcatt 8640
gagtatagtg attttgctac ctctgcttgc gttcttgctg ctgagtgtac aatttttaag 8700
gatgctatgg gcaaacctgt gccatattgt tatgacacta atttgctaga gggttctatt 8760
tcttatagtg agcttcgtcc agacactcgt tatgtcctta tggatggttc catcatacag 8820
tttcctaaca cttacctgga gggttctgtt agagtagtaa caacttttga tgctgagtac 8880
tgtagacatg gtacatgtga aagatcagaa gctggtattt gcttatctac cagtggtaga 8940
tgggttctta acaatgaaca ttatagagct ctacctggag tattttgtgg tgttgatgca 9000
atgaatctta tagcaaacat ctttactccc cttgtgcaac ctgtgggtgc tttagatgtg 9060
tctgcttcag tagtggctgg tggtattatt gccatattgg tgacttgtgc tgcctactac 9120
tttatgaaat tcagacgtgc ttttggtgag tacaaccatg ttgttgctgc taatgcactt 9180
ttgtttttga tgtctttcac tatactctgt ctggcaccag cttatagctt tttgccagga 9240
gtctactcag tcttttactt gtacttgaca ttctatttca ctaatgatgt ttcgttcttg 9300
gctcaccttc aatggtttgc catgttttct cctattgtgc ctttttggat aacagcaatc 9360
tatgtattct gtatttctct gaagcactgc cattggttct ttaacaacta tcttaggaaa 9420
agagtcatgt ttaatggagt tacatttagt accttcgagg aggctgcttt gtgtaccttt 9480
ttgctcaata aggaaatgta cctaaaattg cgtagtgaga cactgttgcc acttacacag 9540
tacaacaggt atcttgctct atataacaag tacaagtatt tcagtggagc cttagatact 9600
accagctatc gtgaagcagc ttgctgccac ttagcaaagg ctctaaatga ctttagcaat 9660
tcaggtgctg atgttctcta ccaaccacca cagacatcaa tcacttctgc tgttctgcag 9720
agtggtttta ggaaaatggc attcccatca ggcaaagttg aagggtgcat ggtacaagta 9780
acctgtggaa ctacaactct taatggattg tggttagatg acacagtata ctgtccaaga 9840
catgtcattt gcacagcaga agacatgctt aatcctaact atgaagatct gctcattcgc 9900
aaatccaacc atagcttcct tgttcaggct ggcaatgtac aactccgagt tatcggccat 9960
tctatgcaaa attgtctgct taggcttaaa gttgatacct ctaaccctaa gacacccaag 10020
tataaatttg tccgtattca acctggtcaa acattttcag ttctagcatg ctacaatggt 10080
tcaccatctg gtgtttatca gtgtgccatg agacctaacc ataccattaa aggttctttc 10140
cttaatggat catgtggtag tgttggtttt aacattgatt atgattgcgt gtctttctgc 10200
tatatgcatc acatggagct ttcaacagga gtacacgctg gtactgactt agaaggtaaa 10260
ttctatggtc catttattga cagacaaact gcccaggcgg caggtacaga cacaaccata 10320
acattaaatg ttttggcatg gctgtatgct gctgttatca atggtgatag gtggtttctt 10380
aatagattca ccactacttt gaatgacttt aaccttgtgg caatgaagta caactatgaa 10440
cctttgacac aagatcatgt tgacatattg ggacctcttt ctgctcaaac gggaattgct 10500
gtcttagata tgtgtgctgc tttgaaagag ctgctgcaga atggtatgaa tggtcgtact 10560
atccttggta gcactatttt agaagatgag tttacaccat ttgatgttgt tagacaatgc 10620
tctggtgtta ccttccaagg taagttcaag aaaattgtta agggcactca tcattggatg 10680
cttttaactt tcttgacatc actattgatt cttgtccaaa gtactcagtg gtcactgttt 10740
ttctttgttt acgagaatgc tttcttgcca tttactcttg gtattatggc aattgctgca 10800
tgtgctatgc tgcttgttaa gcataagcac gcattcttgt gcttgtttct gttaccttct 10860
cttgcaacag ttgcttactt taatatggtc tacatgcctg ccagctgggt gatgcgtatt 10920
atgacatggt ttgaattggc tgacactagc ttgtctggtt atcggcttaa ggactgtgtt 10980
atgtatgctt cagctttagt tttgcttatt ctcatgacag ctcgcactgt ttatgatgat 11040
gctgctagac gtgtttggac actgatgaat gtcattacac ttgtttacaa agtctactat 11100
ggtaatgctt tagatcaagc tatttccatg tgggccttag ttatttctgt aacctctaac 11160
tattctggtg tcgtcacgac tatcatgttt ttagctagag ctatagtgtt tgtgtgtgtt 11220
gagtattacc cattgttatt tattactggc aacaccttac agtgtatcat gcttgtttat 11280
tgtttcttag gctattgttg ctgctgctat tttggccttt tctgtttact caaccgttac 11340
ttcaggctta ctcttggtgt ttatgactat ttggtctcta cacaagaatt taggtatatg 11400
aactcccagg ggcttttgcc tcctaagagt agtattgatg ctttcaagct taacattaag 11460
ttgttgggta ttggaggtaa accatgtatc aaggttgcta ctgtacagtc taaaatgtct 11520
gacgtaaagt gcacatctgt ggtactgctc tcggttcttc aacaacttag agtagagtca 11580
tcttctaaat tgtgggcaca atgtgtacaa ctccacaatg atattcttct tgcaaaagac 11640
acaactgaag ctttcgagaa gatggtttct cttttgtctg ttttgctatc catgcagggt 11700
gctgtagaca ttaacaagtt gtgcgaggaa atgctcgaca accgtgctac tcttcaggct 11760
attgcttcag aatttagttc tttaccatca tatgccgctt atgccactgc ccaagaggcc 11820
tatgagcagg ctgtagctaa tggtgattct gaagtcgttc tcaaaaagtt aaagaaatct 11880
ttgaatgtgg ctaaatctga gtttgaccgt gatgctgcca tgcaacgcaa gttggaaaag 11940
atggcagatc aggctatgac ccaaatgtac aaacaggcaa gatctgagga caagagggca 12000
aaagtaacta gtgctatgca aacaatgctt ttcactatgc ttaggaagct tgataatgat 12060
gcacttaaca acattatcaa caatgcgcgt gatggttgtg ttccactcaa catcatacca 12120
ttgactacag cagccaaact tatggttgtt gtccctgatt atggtaccta caagaacact 12180
tgtgatggta acacctttac atatgcatct gcactctggg aaatccagca agttgttgat 12240
gcagatagta agattgttca acttagtgaa attaacatgg ataattcacc aaatttggct 12300
tggcctctta ttgttacagc tctaagagcc aactcagctg tcaaactaca gaataatgaa 12360
ctgagtccgg tagcactacg acagatgtct tgtgcggctg gtaccacaca aacagcttgt 12420
actgatgaca atgcacttgc ctactataac aattcgaagg gagttaggtt tgtgctggca 12480
ttactatcag accaccaaga tctcaaatgg gctagattcc ctaagagtga tggtacaggt 12540
acaatttaca cagaactgga accaccttgt aggtttgtta cagacacacc aaaagggcct 12600
aaagtgaaat acttgtactt catcaaaggc ttaaataacc taaatagagg tatggttctg 12660
ggcagtttag ctgctacagt acgtcttcag gctggaaatg ctacagaagt acctgccaac 12720
tcaactgtgc tttccttctg tgcctttgca gtagaccctg ctaaagcata taaggattac 12780
ctagcaagtg gaggacaacc aatcaccaac tgtgtgaaga tgttgtgtac acacactggt 12840
acaggacagg caattactgt aacaccagaa gctaacatgg accaagagtc ctttggtggt 12900
gcttcatgct gtctgtattg tagatgccac attgatcatc caaatcctaa aggattttgt 12960
gacttgaaag gtaagtacgt ccaaatacct accacttgtg ctaatgaccc agtgggtttt 13020
acacttagaa acacagtctg taccgtctgc ggaatgtgga aaggttatgg ctgtagttgt 13080
gatcaactcc gcgaacccat gatgcagtct gcggatgcgt caacgttttt aaacgggttt 13140
gcggtgtaag tgcagcccgt cttacaccgt gcggcacagg cactagcact gatgtcgtct 13200
acagggcttt tgatatttac aacgaaaaag ttgctggttt tgcaaagttc ctaaaaacta 13260
attgctgccg cttccaggag aaggatgagg aaggcaattt attagactct tactttgtag 13320
ttaagaggca tactatgtct aactaccaac atgaagagac tatttataac ttggttaaaa 13380
attgtccagc ggttgctgtt catgattttt tcaagtttag agtagatggt gacatggtac 13440
cacatatatc acgtcagcgt ctaactaaat acacaatggc tgatttagtc tatgctctac 13500
gtcactttga cgagggtaat tgtgacacat taaaagaaat actcgtcaca tacaattgtt 13560
gtgatgatga ttattacaat aagaaggatt ggtatgactt cgtagagaat cctgacatct 13620
tacgcgtata tgctaactta ggtgagcgtg tacgccaagc attattaaag actgtacaat 13680
tctgcgatgc tatgcgtgat gcaggcattg taggcgtact gacattagat aatcaggatc 13740
ttaatgggaa ctggtacgat ttcggtgatt tcgtacaagt agcaccaggc tgcggagttc 13800
ctattgtgga ttcatactac tcattgctga tgcccatcct cactctgact agggcattgg 13860
ctgctgagtc ccatatggat gctgatctcg caaaaccact tattaagtgg gatttgctga 13920
aatatgattt tacggaagag agactttgcc tcttcgaccg ttactttaaa tattgggacc 13980
agacatacca tcccaattgt attaactgtt tggatgatag gtgtatcctt cattgtgcaa 14040
actttaatgt gttattttct actgtgtttc cacctacaag ttttggacca ctagtaagaa 14100
aaatatttgt agatggtgtt ccttttgttg tttcaactgg ataccatttt cgtgagttag 14160
gagttgtaca taatcaggat gtaaacttac atagctcgcg tctcagtttc aaggaacttt 14220
tagtgtatgc tgctgatcca gccatgcatg cagcttctgg caatttattg ctagataaac 14280
gcactacatg cttttcggta gctgcactaa caaacaatgt tgcttttcaa actgtcaaac 14340
ccggtaattt taacaaagac ttttatgact ttgctgtgtc taaaggtttc tttaaggaag 14400
gaagttctgt tgaactaaaa cacttcttct ttgctcagga tggcaatgct gctatcagtg 14460
attatgacta ttatcgttat aatctgccaa caatgtgtga tatcagacaa ctcctattcg 14520
tagttgaagt tgttgataaa tactttgatt gttacgatgg tggctgtatt aatgccaacc 14580
aagtaatcgt taacaatctg gacaaatcag ctggtttccc atttaataaa tggggtaagg 14640
ctagacttta ttatgactca atgagttatg aggatcaaga tgcacttttc gcgtatacta 14700
agcgtaatgt catccctact ataactcaaa tgaatcttaa gtatgccatt agtgcaaaga 14760
atagagctcg cactgtagct ggtgtctcta tctgtagtac tatgacaaat agacagtttc 14820
atcagaaatt attgaagtca atagccgcca ctagaggagc tactgtggta attggaacaa 14880
gcaaatttta cggtggctgg cataatatgt taaaaactgt ttacagtgat gtagaaactc 14940
cacaccttat gggttgggat tatccaaaat gtgacagagc catgcctaac atgcttagga 15000
taatggcctc tcttgttctt gctcgcaaac atagcacttg ctgtaactta tcacaccgtt 15060
tctacaggtt agctaacgag tgtgcgcaag tattaagtga gatggtcatg tgtggcggct 15120
cactatatgt taaaccaggt ggaacatcat ccggtgatgc tacaactgct tatgctaata 15180
gtgtctttaa catttgtcaa gctgttacag ccaacgtaaa tgcacttctt tcaactgatg 15240
gtaataagat agctgacaag tatgtccgca atctacaaca caggctctat gagtgtctct 15300
atagaaatag ggatgttgat cacgaattcg tggatgagtt ttacgcttac ctgcgtaaac 15360
atttctccat gatgattctt tctgatgacg ccgttgtgtg ctataacagt aactatgcgg 15420
ctcaaggttt agtagctagc attaagaact ttaaggcagt tctttattat caaaataatg 15480
tgttcatgtc tgaggcaaaa tgttggactg agactgacct tactaaagga cctcacgaat 15540
tttgctcaca gcatacaatg ctagttaaac aaggagatga ctacgtgtac ctgccttacc 15600
cagatccatc aagaatatta ggcgcgggct gttttgtcga tgatattgtc aaaacagatg 15660
gtacacttat gattgagagg tttgtgtcat tagctattga tgcctacccc cttactaaac 15720
atcctaatca ggagtatgct gatgtctttc acttgtattt acaatacatt aggaagttac 15780
atgatgagct tactggtcac atgctagaca tgtattctgt aatgctaacc aatgacaaca 15840
cctcacggta ttgggaacct gagttttatg aagctatgta cacaccacac acagtcttgc 15900
aggctgtagg tgcttgtgta ttgtgtaatt cacagacttc acttcgttgc ggtgcctgca 15960
ttaggagacc attcctgtgc tgcaagtgct gctatgacca tgtcatttca acatcacata 16020
aattagtgtt gtctgttaat ccctatgttt gcaatgcacc aggttgtgat gtcactgacg 16080
tgacacaact atatctagga ggtatgagct attactgcaa gtcacataag cctcccatta 16140
gttttccatt gtgtgctaat ggtcaggttt ttggtttata caagaacaca tgtgtaggta 16200
gtgacaatgt cactgacttc aatgcgatag caacatgtga ttggactaat gctggcgatt 16260
acatacttgc caacacttgt actgagagac tcaaactctt tgcagcagaa acactcaaag 16320
ctactgagga aacattcaag ctgtcatatg gtattgccac tgtacgtgaa gtactctctg 16380
acagagaatt gcatctttca tgggaggttg gaaaacctag accaccattg aatagaaatt 16440
atgtctttac tggttaccgt gtaactaaaa atagtaaagt acagattgga gagtacacct 16500
ttgaaaaagg tgactatggt gatgctgttg tgtacagagg tactacgaca tacaaattga 16560
atgttggtga ttactttgtg ttgacatctc acactgtaat gccacttagt gcacctactc 16620
tagtgccaca agagcactat gtgagaatta ctggcttgta cccaacactc aacatctcag 16680
atgagttttc tagcaatgtt gcaaattatc aaaaggtcgg tatgcaaaag tactctacac 16740
tccagggccc acctggcact ggtaagagtc attttgccat cggacttgct ctctactacc 16800
catctgctcg catagtgtat acagcttgct cccatgcagc tgttgatgcc ctatgcgaaa 16860
aggcactaaa atacttgccc atagataaat gtagtagaat catacctgcg cgtgctcgcg 16920
tagagtgttt tgacaaattc aaagtgaatt caacactaga acagtatgtt ttctgcactg 16980
taaatgcatt gccagaaaca actgctgata ttgtagtctt tgatgaaatc tctatggcta 17040
ctaattatga cttgagtgtt gtcaatgcta gacttcgtgc aaaacactac gtctatattg 17100
gcgatcctgc tcaattacca gccccgcgca cattgctgac caaaggcaca ctagaaccag 17160
aatacttcaa ttcagtgtgc agacttatga aaacaatagg tccagacatg ttccttggaa 17220
cttgtcgccg ttgtcctgct gaaattgtcg acactgtgag tgctttagtt tatgataata 17280
agctaaaggc acacaaggag aagtcagctc aatgtttcaa aatgttttat aagggtgtta 17340
ttacacatga tgtttcatct gcaattaaca gacctcaaat aggcgttgta agagaatttc 17400
ttacacgcaa tcctgcttgg agaaaagctg tttttatctc accttataat tcacagaatg 17460
ctgtagcttc aaaaatctta ggattgccta cgcagactgt cgattcctcc cagggttctg 17520
agtatgacta tgtcatattc acacaaacta ctgaaacagc acactcttgc aatgttaacc 17580
gctttaatgt ggctatcaca agagcaaaaa ttggcatttt gtgcataatg tccgatagag 17640
atctttatga caaactgcaa ttcacaagtt tagaagtacc acgccgcaat gtggctacat 17700
tacaggcaga aaatgtaact ggacttttta aggactgtag taagatcatc accggtcttc 17760
atccaacaca ggcacctaca cacctcagcg ttgatacaaa atttaagact gagggactat 17820
gtgttgacat accaggcata ccaaaggaca tgacctaccg tagactcatc tctatgatgg 17880
gtttcaaaat gaattaccaa gttaatggtt accctaatat gtttattacc cgtgaggaag 17940
ctattcgtca cgttcgtgca tggattggct tcgacgtaga gggctgtcat gcaactagag 18000
atgctgtggg tactaaccta cctctccagc taggattttc tacaggtgtt aacttagtag 18060
ctgtaccgac tggctatgtt gacactgaaa ataacacaga attcaccaga gttaatgcaa 18120
gacctccacc aggtgaccag tttaaacatc ttataccact catgtacaaa ggcttgccct 18180
ggaatgtagt gcgtattaag atagtacaaa tgctcagtga tacactgaaa ggattgtcag 18240
acagagtcgt gtttgtcctt tgggcgcatg gctttgagct tacatcaatg aagtactttg 18300
tcaagattgg acctgaaaga acgtgctgtc tgtgtgacaa acgtgcaact tgcttttcta 18360
cttcatcaga tacttatgcc tgctggaatc attctgtggg ttttgactat gtctacaacc 18420
catttatgat tgatgtccag cagtggggtt ttacgggtaa ccttcagagt aaccacgacc 18480
aacattgtca agtgcatgga aatgcacatg tggctagttg tgatgctatc atgactagat 18540
gcttggcggt ccatgagtgc tttgttaagc gcgttgattg gtctgttgaa taccccatta 18600
taggagatga actgaagatt aattccgctt gcagaaaagt acagcatatg gttgtaaagt 18660
ctgcattgct tgctgataag tttccagttc ttcatgacat tggaaatcca aaggctatta 18720
agtgtgtgcc tcaggcggaa gtagaatgga agttctatga tgctcagcca tgcagtgaca 18780
aagcctataa aatagaggaa cttttctatt cctacgctac acatcatgat aaattcactg 18840
atggtgtttg tttgttttgg aactgtaacg ttgatcgtta cccagccaat gcaattgtat 18900
gtaggtttga cacgagagtt ttgtcaaact tgaatttacc aggttgtgac ggtggtagtt 18960
tgtatgtgaa taagcatgca ttccacactc cagcttttga taaaagtgca tttaccaatt 19020
taaagcaatt gcctttcttt tattattctg acagtccttg tgagtctcat ggcaaacaag 19080
tagtgtcaga tattgattat gtaccactta aatctgctac gtgtattaca cgatgcaatt 19140
tgggaggtgc tgtttgcaga caccatgcaa atgagtaccg acagtactta gatgcatata 19200
acatgatgat ttctgctgga tttagcctat ggatttacaa acagtttgat acttataatc 19260
tgtggaatac attcaccagg ttacagagtt tagaaaatgt ggcttacaat gttgttaaca 19320
aaggacactt cgatggacaa gctggtgaag cacctgtttc catcattaat aatgctgttt 19380
acacaaaggt agatggtgtt gatgtagaga tctttgaaaa caagacaaca cttcctgtta 19440
atgttgcgtt tgagctttgg gctaagcgta acattaaacc agtgccagag attaagatac 19500
tcaataactt gggtgtcgat atcgctgcta atactgtggt ctgggactac aagagagaag 19560
caccagcaca tatgtcaaca ataggtgtct gtacaatgac cgacattgct aagaaaccta 19620
ctgagagtgc ttgttcctcg cttactgtct tatttgatgg tagagtggaa ggacaggtag 19680
acctttttag aaatgcccgt aatggtgttt taataacaga aggttcagtt aaaggtttaa 19740
caccttcaaa ggggccagca caagctagtg tcaatggagt cacattaatt ggagaatccg 19800
taaaaacaca gtttaactat tttaagaaag tagatggcat tattcaacag ttgcctgaaa 19860
cctactttac tcagagccga gacttagagg atttcaagcc cagatcacaa atggaaactg 19920
actttcttga gctcgctatg gatgaattca tacaacggta taagctagag ggttatgcct 19980
tcgaacatat cgtttatggg gatttcagtc atggacaact tggcggcctt catctaatga 20040
ttggtttagc caagcgctca caagattcac cgcttaaatt agaggacttt atccctatgg 20100
atagcacagt gaaaaattac ttcataacag atgcacaaac aggttcatca aaatgtgtgt 20160
gctctgttat tgacctttta cttgatgact ttgttgagat aataaagtca caggatttgt 20220
cagtaatttc taaggtagtc aaggttacaa ttgactacgc tgagatttca ttcatgcttt 20280
ggtgtaaaga tggacacgtt gaaaccttct accctaaatt acaagcaagt caagcgtggc 20340
aaccgggggt tgcaatgcct aacttgtaca agatgcaaag aatgcttctt gaaaagtgtg 20400
accttcagaa ttatggtgaa aatgctgtca taccaaaagg aataatgatg aatgtcgcaa 20460
aatacactca actgtgtcaa tacttaaata cacttacttt agctgtaccc tacaacatga 20520
gagttattca ctttggtgct ggatctgata aaggagttgc accaggtaca gctgtactca 20580
ggcaatggtt gccaactggc acactacttg tcgattcaga ccttaatgac ttcgtctctg 20640
acgcggattc tactttaatt ggagactgtg caacagtaca tacggctaat aaatgggatc 20700
tcattattag cgatatgtat gaccctaaga caaaacatgt gacaaaagag aacgactcaa 20760
aagaagggtt tttcacttac ctgtgtggat ttataaagca aaagctagcc ttgggtggct 20820
ctgtggctgt gaagataaca gagcattctt ggaatgctga tctttacaag cttatgggac 20880
acttctcatg gtggacagct tttgttacaa atgtaaatgc atcatcatca gaagcatttc 20940
taattggggt taactatcta ggcaagccaa aggaacaaat tgatggctat accatgcatg 21000
ctaactacat cttttggagg aacacaaatc ctattcaatt gtcttcctat tcactttttg 21060
acatgagcaa gtttcctctc aaattaagag ggactgctgt tatgtattta aaggagaatc 21120
agatcaatga catgatttat tctctacttg agaaaggtag acttatcatt agagagagta 21180
acaaagttgt ggtgtctagt gatattttag ttaataacta aacgaacatg aaattgttag 21240
ttttagtttt tgctactcta gtctcctctt acactataga gaagtgcctt gattttgatg 21300
accgcactcc acctgcaaat actcaatttt tatcttctca cagaggtgtt tattacccag 21360
atgacatttt taggtctaat gtcttgcatt tagtacaaga tcatttccta ccttttgact 21420
ctaacgtcac caggtttata acgtttggcc taaattttga taatcccata atacccttca 21480
gggatggtat ttattttgct gcgactgaaa agtctaatgt tattagagga tgggtttttg 21540
gttctacaat gaacaacaaa tctcaatccg ttataataat gaacaactca actaatttag 21600
tcattagggc ttgtaatttt gagttgtgtg acaatccatt ttttgttgtg ttgaaatcta 21660
acaacactca aataccatct tacatattta ataatgcatt caattgcaca tttgaatatg 21720
tttctaagga ttttaaccta gaccttggtg aaaaaccagg taatttcaag gatctcagag 21780
agtttgtttt caggaataaa gatggttttt tgcatgttta ttccggttac caacccattt 21840
ctgctgccag tggtttgcca actggtttta atgcactcaa acctattttc aagttacctc 21900
tgggtattaa tattactaat ttcagaacac ttctgactgc ttttccgcct agacctgatt 21960
attggggtac ttcagctgca gcttattttg taggatattt aaaaccaact acattcatgc 22020
tcaagtatga tgaaaatggt acaatcacag atgctgtcga ttgttctcaa aatccacttg 22080
ctgaactcaa atgctctgtt aaaagttttg agattgacaa aggaatttat caaacctcca 22140
attttagggt agcaccctca aaggaagttg tgaggttccc taatattaca aacctgtgtc 22200
cttttgggga ggtttttaat gctactacat ttccttctgt ctatgcatgg gagaggaaaa 22260
gaatttctaa ttgtgttgct gattactctg tactctacaa ctcaacatct ttttcaactt 22320
ttaagtgtta tggcgtttct gccactaagc tgaatgacct ttgcttctcc aacgtctatg 22380
cagattcatt cgtagtcaaa ggagatgatg taaggcaaat agcaccagga cagaccggtg 22440
ttattgctga ttataattac aaattaccag atgacttctt gggttgtgtc ctagcatgga 22500
acaccaattc taaagattct tccacttccg gtaattataa ttatttatat agatgggtta 22560
gaaggtctaa gcttaaccct tatgagcgcg acttatctaa cgacatctat tcacctggag 22620
gtcagtcttg ctcagctgta ggtcctaatt gttataaccc cttacgtcca tatggctttt 22680
ttacaacagc tggtgttgga taccaacctt atagagttgt agtactttct tttgaacctt 22740
taaatgcacc cgctacagtc tgtggaccaa aattatccac cgaccttatt aaaaatcaat 22800
gtgtcaattt taactttaat ggactcactg gtactggtgt gttaactcct tcttcaaaga 22860
gatttcaacc atttcaacaa tttggtcgtg atgtttcgga tttcactgat tcagttcgag 22920
atccgaagac gtctgaaata ttagacattt caccttgctc ttttggcggt gtaagtgtaa 22980
tcacacctgg aacaaatact tcatcagaat ttgctgttct atatcaagat gttaactgca 23040
ctgatgttcc tgtagcaatc catgcagacc aactcacacc ttcttggcgc gtatactcta 23100
ctggaaataa tgtatttcaa acccaggcag gctgtcttat aggagctgag catgtcgaca 23160
cttcttatga gtgcgacatt cctattggag ctggcatttg tgctagttac catacagttt 23220
cttcattacg tagtactagc caaaaatcta ttgtggctta tactatgtct ttaggtgctg 23280
atagttcaat tgcttactct aataacacca ttgctatacc tactaacttt tcaattagca 23340
ttactacaga agtaatgcct gtttctatgg ctaaaacctc tgtagattgt aatatgtaca 23400
tctgcggaga ttctactgaa tgtgctaatt tgcttctcca atatggtagc ttttgcacac 23460
aactaaatcg tgcactctca ggtattgctg ttgaacagga tcgcaacaca cgtgaagtgt 23520
tcgctcaagt caaacaaatg tacaaaaccc caactttgaa agattttggt ggttttaatt 23580
tttcacaaat attacctgac cctctaaagc caactaagag gtcttttatt gaggacttgc 23640
tctttaataa ggtgacactc gctgatgctg gctttatgaa gcaatatggc gaatgcctag 23700
gtgatattaa tgctagagat ctcatttgtg cgcagaagtt caatggactt acagtgctgc 23760
cacctctgct cactgatgat atgattgctg cctacactgc tgctctagtt agtggtactg 23820
ccactgctgg atggacattc ggtgctggcg ctgctcttca aatacctttt gctatgcaaa 23880
tggcatatag gttcaatggc attggagtta ctcaaaatgt tctctatgag aaccaaaaac 23940
aaatcgccaa tcaatttaac aaggcgatca gccaaattca agaatcactc acaacaacat 24000
ccactgcatt gggcaagctg caagatgtcg tcaaccagaa tgctcaagca ttaaacacac 24060
ttgttaaaca acttagctcc aattttggtg cgatttcaag tgtgctaaat gatatccttt 24120
cgcgacttga taaagtcgag gcagaggtac aaattgacag gttaattaca ggcagactgc 24180
aaagccttca aacctatgta acacaacaac taatcagggc tgctgaaatc agggcttctg 24240
ctaatcttgc tgctactaaa atgtctgagt gtgttcttgg acaatcaaaa agagttgact 24300
tttgcggaaa gggctaccat cttatgtcct tcccacaagc agccccgcat ggtgttgtct 24360
tcctacatgt cacatatgtg ccatctcaag agagaaactt caccacagcg ccagcaattt 24420
gtcatgaagg caaagcatac ttccctcgtg aaggtgtttt tgtgtttaat ggcacttcgt 24480
ggtttattac acagaggaac ttcttttctc cacaaataat tactacagac aatacatttg 24540
tctccggaag ttgtgatgtc gtaattggca tcattaacaa cacagtttat gatcctctgc 24600
aacctgagct tgactcattc aaagaagagc tggacaagta cttcaaaaat cacacatcac 24660
cagatgttga tcttggcgac atttcaggca ttaacgcttc tgtcgtcaac attcaaaaag 24720
aaattgaccg cctcaatgag gtcgctaaaa atttaaatga atcacgcatt gaccttcaag 24780
aattgggaaa atatgagcaa tatattaaat ggccttggta tgtttggctc ggcttcattg 24840
ctggactaat tgccatcgtc atggttacaa tcttgctttg ttgcatgact agttgttgca 24900
gttgcctcaa gggtgcatgc tcttgtggtt cttgctgcaa gtttgatgag gatgactctg 24960
agccagttct caagggtgtc aaattacatt acacataaac gaacttatgg atttgtttat 25020
gagaattttt actcttggac caattactgc acagccagga aaaattgaca atgcttctcc 25080
tgcaagtact gttcatgcta cagcaacgat accgctacaa gcctcactcc ctttcggatg 25140
gcttgttatt ggcgttgcat ttcttgctgt ttttcagagc gctaccaaaa taatttcgct 25200
caataaaaga tggcagctag ccctttataa gggcttccag ttcatttgca atttactgct 25260
gctatttgtt accatctatt cacatctttt gcttgtcgct gcaggtatgg aggcgcaatt 25320
tttgtacctc tatgccttga tatattttct acaatgcatc aacgcatgta gaattatcat 25380
gagatgttgg ctttgttgga agtgcaaatc caagaaccca ttactttatg atgccaacta 25440
ctttgtttgc tggcacacac ataactatga ctactgtata ccgtataaca gtgtcacaga 25500
tacaattgtc gttactgcag gtgacggcat ttcaacacca aaactcaaag aagactacca 25560
aattggtggt tattctgaga attggcactc aggtgttaaa gactatgtcg ttgtacatgg 25620
ctatttcacc gaagtttact accagcttga gtctacacaa attactacag acactggtat 25680
tgaaaatgct acattcttca tctttaacaa gcttgttaaa gacccaccga atgtgcaaat 25740
acacacaatc gacggctctt caggagttgt aaatccagca atggatccaa tttatgatga 25800
gccgacgacg actactagcg tgcctttgta agcacaagaa agtgagtacg aacttatgta 25860
ctcattcgtt tcggaagaaa caggtacgtt aatagttaat agcgtacttc tttttcttgc 25920
tttcgtggta ttcttgctag tcacactagc catccttact gcgcttcgat tgtgtgcgta 25980
ctgctgcaat attgttaacg tgagtttagt aaaaccaact gtttacgttt actcgcgtgt 26040
taaaaatctg aactcttctg aaggagttcc tgatcttctg gtctaaacga actaactatt 26100
attattattc tgtttggaac tttaacattg cttatcatgg ctgagaacgg gattatttcc 26160
gttgaggagc ttaaaagact cctggaacaa tggaacctag taataggttt cctattccta 26220
gcctggatta tgttactaca atttgcctat tctaatcgga acaggttttt gtacataata 26280
aagcttgttt tcctgtggct cttgtggcca gtaacacttg cttgctttgt gcttgctgct 26340
gtttacagaa ttaattgggt gactggcggg attgcgattg caatggcttg tattgtaggc 26400
ttgatgtggc ttagctactt cattgcttcc ttcaggctat ttgctcgtac ccgctcaatg 26460
tggtcattca acccagaaac aaacattctt ctcaatgtgc ctcttcgagg gacaattgtg 26520
accagaccgc tcatggaaag tgaacttgtc attggcgctg tgatcattcg tggtcacttg 26580
cgcatggctg gacactccct agggcgctgt gacatcaagg acctgccaaa agagatcact 26640
gtggctacat cacgaacgct ttcttattac aaattaggag cgtcgcagcg tgtaggcact 26700
gattcaggtt ttgctgcata caaccgctac cgtattggaa actacaaatt aaatacagac 26760
cacgccggta gcaacgacaa tattgctttg ctagtacagt aagtgacaac agatgtttca 26820
tcttgttgac ttccaggtta caatagcaga gatattgatt atcattatga ggactttcag 26880
gattgctatt tggaatcttg acatgataat aagttcaata gtgagacaat tatttaagcc 26940
tctaactaag aataaatatt cagagttaga tgatgaagaa cctatggaga tagattatcc 27000
ttgataaacg aaccactatg ttacttttag taacattgtt tggtttagca tcagggtgca 27060
gcttaccact tacggttagc tgccctagag ccctaccttt cactctacag attaacacta 27120
ctagtgttac tgtggagtgg tatcgggtat ctcctgcatc aatgcaaggt cttacaaaga 27180
taaatactgg cagcactatt tttgataaca actttagtgt agtcaataat aatttgtact 27240
tcaaacagtg ttttggaggc ttctttttac agcacgctgt taccgccagg gtaagcatga 27300
cggtgctata ttagataatt ctcaacctgt ctttgtggat gctaggaatt atgtaccaac 27360
tactgcacta ttagtctcat cgcagggcat tgtgcagcca aaaagttcca atgtgttagc 27420
tatag 27425
<210> 3
<211> 46
<212> DNA
<213> FA(Artificial Sequence)
<400> 3
tgaccgcggc taatacgata ttaggttttt acctacccag gaaaag 46
<210> 4
<211> 35
<212> DNA
<213> F-c1575a(Artificial Sequence)
<400> 4
caggccatac tggaattact ggtgacaacg tggag 35
<210> 5
<211> 39
<212> DNA
<213> R-c1575a (Artificial Sequence)
<400> 5
gtcaccagta attccagtat ggcctgaacc aatatcagc 39
<210> 6
<211> 23
<212> DNA
<213> RA(Artificial Sequence)
<400> 6
atggtggcca ttatggctct aac 23
<210> 7
<211> 25
<212> DNA
<213> FB(Artificial Sequence)
<400> 7
tgttagagcc ataatggcca ccatc 25
<210> 8
<211> 37
<212> DNA
<213> RB(Artificial Sequence)
<400> 8
ctgtagccac gagggcccta agtttttcca tagggac 37
<210> 9
<211> 34
<212> DNA
<213> FC(Artificial Sequence)
<400> 9
cttagggccc tcgtggctac agctcatagc gagc 34
<210> 10
<211> 27
<212> DNA
<213> RCm(Artificial Sequence)
<400> 10
acctgccgcc tgggcagttt gtctgtc 27
<210> 11
<211> 25
<212> DNA
<213> FCm(Artificial Sequence)
<400> 11
ctgcccaggc ggcaggtaca gacac 25
<210> 12
<211> 23
<212> DNA
<213> RC(Artificial Sequence)
<400> 12
gctcataggc ctcttgggca gtg 23
<210> 13
<211> 23
<212> DNA
<213> FD(Artificial Sequence)
<400> 13
cactgcccaa gaggcctatg agc 23
<210> 14
<211> 32
<212> DNA
<213> RD(Artificial Sequence)
<400> 14
cagtgccagg tgggccctgg agtgtagagt ac 32
<210> 15
<211> 34
<212> DNA
<213> FE(Artificial Sequence)
<400> 15
agggcccacc tggcactggt aagagtcatt ttgc 34
<210> 16
<211> 35
<212> DNA
<213> RE(Artificial Sequence)
<400> 16
acctcgccaa aagggcacag gtttgtaata ttagg 35
<210> 17
<211> 37
<212> DNA
<213> FF(Artificial Sequence)
<400> 17
acctgtgccc ttttggcgag gtttttaatg ctactac 37
<210> 18
<211> 24
<212> DNA
<213> RF(Artificial Sequence)
<400> 18
aggtaggcct ctagggcagc taac 24
<210> 19
<211> 25
<212> DNA
<213> FG(Artificial Sequence)
<400> 19
tagctgccct agaggcctac ctttc 25
<210> 20
<211> 54
<212> DNA
<213> RG(Artificial Sequence)
<400> 20
actggcgcgc cttttttttt tttttttttt ttttttgtca ttctcctgag aagc 54
<210> 21
<211> 45
<212> DNA
<213> R-EsBsaI(Artificial Sequence)
<400> 21
actggtctct tcgtttagtt attaactaaa atatcactag acacc 45
<210> 22
<211> 33
<212> DNA
<213> F-FsBsaI(Artificial Sequence)
<400> 22
tgaggtctcc gaacttatgg atttgtttat gag 33
<210> 23
<211> 45
<212> DNA
<213> F- SHC014(Artificial Sequence)
<400> 23
agtggtctca acgaacatga aattgttagt tttagttttt gctac 45
<210> 24
<211> 44
<212> DNA
<213> R- SHC014(Artificial Sequence)
<400> 24
tcaggtctca gttcgtttat gtgtaatgta atttgacacc cttg 44
<210> 25
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 25
aacaactccg ccccattgac gc 22
<210> 26
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 26
actgcaacac ggacgaaacc gt 22
<210> 27
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 27
ccctggacaa ggatgtgctg gtt 23
<210> 28
<211> 24
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 28
cgacgggact ctccaaagtg tggt 24
<210> 29
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 29
tggcgcgctt catctctact ttg 23
<210> 30
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 30
gccaagatct ctaggcgtca tgttt 25
<210> 31
<211> 25
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 31
tgcatcacat ggagcttcca acagg 25
<210> 32
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 32
acagcagcat acagccatgc ca 22
<210> 33
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 33
tgctatccat gcagggtgct gta 23
<210> 34
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 34
tggaacacaa ccatcacgcg ca 22
<210> 35
<211> 26
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 35
gggaggttgg aaaacctaga ccacca 26
<210> 36
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 36
ctttggtcag caatgtgcgc gg 22
<210> 37
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 37
agccaccaca ttttcaccga ggc 23
<210> 38
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 38
agaaggcaca gctcccttag cca 23
Claims (2)
1. A BALB/c mouse adapted strain of SARS-associated coronavirus WIV1-RsSHC014S, characterized by: the full-length gene sequence is shown as SEQ ID NO: 2, respectively.
2. Use of the BALB/c mouse adapted strain of SARS-associated coronavirus WIV1-RsSHC014S of claim 1 in the preparation of neutralizing antibodies, vaccines, detection reagents and therapeutic drugs.
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| CN202210083738.3A CN114107232B (en) | 2022-01-25 | 2022-01-25 | BALB/c mouse adapted strain of SARS related coronavirus WIV1-RsSHC014S and application thereof |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111961653A (en) * | 2020-08-27 | 2020-11-20 | 华中农业大学 | Novel coronavirus mouse adaptive strain SARS-CoV-2/WBP-1 and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111961653A (en) * | 2020-08-27 | 2020-11-20 | 华中农业大学 | Novel coronavirus mouse adaptive strain SARS-CoV-2/WBP-1 and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| "Understanding the Risk of Bat Coronavirus Emergence"-a Merogenomics" NIH grant review;Dr.M.Raszek;《https://merogenomics.ca/blog/en/145/Understanding-the-Risk-of-Bat-Coronavirus-Emergence-a-Merogenomics-NIH-grant-review》;20210510;全文 * |
| Bat Severe Acute Respiratory Syndrome-Like Coronavirus WIV1 Encodes an Extra Accessory Protein, ORFX, Involved in Modulation of the Host Immune Response;Lei-Ping Zeng et al.;《Journal of Virology》;20160511;第90卷(第14期);第6573-6582页 * |
| Cross-neutralization of SARS coronavirus-specific antibodies against bat SARS-like coronaviruses;Lei-Ping Zeng等;《Science China(Life Sciences)》;20171205;第60卷(第12期);第1399-1402页 * |
| Lei-Ping Zeng等.Cross-neutralization of SARS coronavirus-specific antibodies against bat SARS-like coronaviruses.《Science China(Life Sciences)》.2017,第60卷(第12期),第1399-1402页. * |
| Understanding the Risk of Bat Coronavirus Emergence;匿名;《https://www.documentcloud.org/app?q=WIV1-RsSHC014》;20211021;全文 * |
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