CN113249444A - DNA probe library for hybridization with FGFR1, 2 or 3 gene and application - Google Patents

Abstract

The invention relates to a DNA probe library for hybridization with 1, 2 or 3 genes of Fibroblast Growth Factor Receptors (FGFR) and application thereof, belonging to the technical field of biological medicines. The invention provides a DNA probe library for hybridization with FGFR1, FGFR2 or FGFR3 genes, wherein the DNA probes comprise all the probes listed in SEQ ID NO. 1-186. The invention provides a method for enriching FGFR1, FGFR2 or FGFR3 gene fragments by using a DNA probe library and detecting related fusion variation by combining with a next generation sequencing technology (NGS). By using the gene enrichment method and the specific DNA probe library obtained by screening, the fusion variation in FGFR1, FGFR2 or FGFR3 genes can be accurately obtained by applying the next generation sequencing technology. Has the advantages of large detection flux, high accuracy, short period and high sensitivity.

Description

DNA probe library for hybridization with FGFR1, 2 or 3 gene and application

Technical Field

The invention relates to a DNA probe library for hybridization with 1, 2 or 3 genes of Fibroblast Growth Factor Receptors (FGFR) and application thereof, belonging to the technical field of biological medicines.

Background

The Fibroblast Growth Factor Receptor (FGFR) family mainly includes 4 types of receptors FGFR1, FGFR2, FGFR3 and FGFR4, which play important roles in angiogenesis, wound healing and embryonic development. FGFR is a transmembrane protein, with an extracellular domain, a transmembrane domain, and an cytoplasmic tyrosine kinase domain. The signaling of the FGF-FGFR pathway is tightly controlled by feedback regulation. Mitogen-activated protein kinase phosphatase and Sprouty protein are upregulated after FGFR stimulation and antagonize FGF-dependent activation of extracellular signal-regulated kinase. Strong genetic and functional evidence suggests that FGFR dysregulation can lead to the development and progression of cancer. Genetic alterations of FGFR1, FGFR2 and FGFR3 have been found and reported in many tumor types (Knight V, Cook SJ. De-regulated FGF receptors as therapeutic targets in Cancer. Pharmacol Ther 2010; 125: 105-.

The gene fusion belongs to one kind of gene variation, and is one kind of chimeric gene comprising genes in different positions on two or more chromosomes connected end to end owing to chromosome structure aberration, including deletion, duplication, inversion, translocation, etc. and its expression product is fusion protein. In gene fusion, the sequences of all or a part of two genes are fused to each other to form a novel gene. Gene fusions may be the result of chromosomal translocations, intermediate deletions, or chromosomal inversions. Some kinase fusion is one of the factors inducing cancer, and the inhibitor aiming at FGFR fusion has been approved as a medicament, including the FGFR2 inhibitor Pemazyre (pemigatinib) and the irreversible FGFR oral inhibitor futibatinib.

Traditional methods for detecting gene fusion are FISH and IHC, but these methods are low in throughput and require separate design of kits for individual genes, and fused partner genes are often difficult to detect, thus also having limitations in clinical practice. The technical field needs a sequencing detection scheme with high throughput, high precision and high sensitivity, which can rapidly detect related fusion variation of FGFR1, FGFR2 or FGFR3 genes.

Disclosure of Invention

The invention aims to solve the technical problem of how to rapidly detect related fusion variation of FGFR1, FGFR2 or FGFR3 genes with high flux, high precision and high sensitivity.

In order to solve the above problems, the present invention provides a DNA probe library for hybridizing with FGFR1, FGFR2 or FGFR3 genes; the DNA probe comprises sequences shown in SEQ ID NO.1 to SEQ ID NO. 186.

The invention provides a method for enriching and detecting FGFR1, FGFR2 or FGFR3 gene fragments, which comprises the following steps:

step 1: extracting genomic DNA from a human cell, plasma, tissue or FFPE sample, thereby obtaining fragmented double-stranded DNA as a DNA sample library;

step 2: designing a DNA probe which is hybridized with FGFR1, FGFR2 or FGFR3 gene as a DNA probe library aiming at the FGFR1, FGFR2 or FGFR3 gene to be enriched; the DNA probe library is the DNA probe library of claim 1;

and step 3: hybridizing the DNA sample library with the DNA probe library;

and 4, step 4: separating the hybridization product of step 3, and then releasing the gene fragments enriched by hybridization;

and 5: and (3) carrying out detection of fusion variation on the enriched FGFR1, FGFR2 or FGFR3 gene fragments.

Preferably, the DNA sample pool in the above step 1 is composed of double-stranded DNA fragments, and the above step 1 includes extracting whole genomic DNA of the subject and then fragmenting it; wherein the subject is human and the whole genomic DNA is extracted from a cell, plasma, tissue or FFPE sample of the subject.

Preferably, the fusion mutation detection of the enriched FGFR1, FGFR2 or FGFR3 gene fragments in the step 5 is performed by using next generation sequencing technology. (the high throughput Sequencing technology is also called Next Generation Sequencing; Next Generation Sequencing, NGS for short).

The invention provides an application of a DNA probe library hybridized with FGFR1, FGFR2 or FGFR3 genes in related fusion mutation detection of FGFR1, FGFR2 or FGFR3 genes; the application comprises the method described above.

The invention provides a kit for enriching FGFR1, FGFR2 or FGFR3 gene fragments, which comprises the DNA probe library.

The invention provides a kit for detecting FGFR1, FGFR2 or FGFR3 gene fragment fusion variation, which comprises the DNA probe library.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a DNA probe library for hybridization with FGFR1, FGFR2 or FGFR3 genes, wherein the DNA probes comprise all the probes listed in SEQ ID NO. 1-186. In addition, the invention provides a method for enriching FGFR1, FGFR2 or FGFR3 gene fragments by using the DNA probe library and detecting related fusion variation by combining with next generation sequencing technology (NGS).

The gene enrichment method and the specific DNA probe library obtained by screening are used for applying the next generation sequencing technology; and (3) sequencing the FGFR1, the FGFR2 or the FGFR3 gene fragment, and further accurately obtaining the fusion mutation in the FGFR1, the FGFR2 or the FGFR3 gene. Because the next generation sequencing technology is adopted, the detection flux is large, the accuracy is high, the period is short, and the sensitivity is high.

Drawings

Fig. 1 is a block diagram of a method for capturing a specific FGFR1, FGFR2 or FGFR3 gene fragment based on hybridization selection according to the present invention.

Detailed Description

In order to make the invention more comprehensible, preferred embodiments are described in detail below with reference to the accompanying drawings:

the invention provides a DNA probe library for hybridization with FGFR1, FGFR2 or FGFR3 genes; the DNA probe comprises sequences shown in SEQ ID NO.1 to SEQ ID NO. 186.

The invention provides a method for enriching and detecting FGFR1, FGFR2 or FGFR3 gene fragments, which comprises the following steps:

step 1: extracting genomic DNA from a human cell, plasma, tissue or FFPE sample, thereby obtaining fragmented double-stranded DNA as a DNA sample library;

step 2: designing a DNA probe which is hybridized with FGFR1, FGFR2 or FGFR3 gene as a DNA probe library aiming at the FGFR1, FGFR2 or FGFR3 gene to be enriched; the DNA probe library is the DNA probe library of claim 1;

and step 3: hybridizing the DNA sample library with the DNA probe library;

and 4, step 4: separating the hybridization product of step 3, and then releasing the gene fragments enriched by hybridization;

and 5: and (3) carrying out detection of fusion variation on the enriched FGFR1, FGFR2 or FGFR3 gene fragments.

The DNA sample pool in the above step 1 is composed of double-stranded DNA fragments, and the above step 1 comprises extracting whole genomic DNA of the subject and then fragmenting it; wherein the subject is human and the whole genomic DNA is extracted from a cell, plasma, tissue or FFPE sample of the subject.

And (3) in the step 5, the fusion mutation detection of the enriched FGFR1, FGFR2 or FGFR3 gene fragments adopts a next generation sequencing technology. (the high throughput Sequencing technology is also called Next Generation Sequencing; Next Generation Sequencing, NGS for short).

The invention provides an application of a DNA probe library hybridized with FGFR1, FGFR2 or FGFR3 genes in related fusion mutation detection of FGFR1, FGFR2 or FGFR3 genes; the application comprises the method described above.

The invention provides a kit for enriching FGFR1, FGFR2 or FGFR3 gene fragments, which comprises the DNA probe library.

The invention provides a kit for detecting FGFR1, FGFR2 or FGFR3 gene fragment fusion variation, which comprises the DNA probe library.

The present invention provides a method for capturing specific FGFR1, FGFR2 or FGFR3 gene fragments based on hybridization selection. Specifically, the method of the present invention comprises: extracting genomic DNA from a human cell, plasma, tissue or FFPE sample, thereby obtaining fragmented double-stranded DNA as a DNA sample library; in addition, a DNA probe which hybridizes with the FGFR1, FGFR2 or FGFR3 gene is designed as a DNA probe library aiming at the FGFR1, FGFR2 or FGFR3 gene to be enriched; then, hybridizing the DNA sample library with the DNA probe library, thereby enriching the FGFR1, FGFR2 or FGFR3 gene fragments from the DNA sample library. After the adaptive treatment, the fusion mutation detection of the FGFR1, FGFR2 or FGFR3 gene fragments can be carried out by adopting a next generation sequencing technology.

The high throughput Sequencing technology is also called Next Generation Sequencing (NGS), and can sequence hundreds of thousands to millions of molecules in a cell genome at a time, so that the technology is widely applied to genome research, is also an important means for gene detection of tumor patients, can be used for gene mutation detection of different individuals such as tumor patients, and provides beneficial suggestions and guidance for individualized treatment. NGS is a method for detecting and fusing high flux, high precision and high sensitivity.

Example 1: enriching FGFR1, FGFR2, or FGFR3 genes of tissue/FFPE samples and detecting relevant fusion variants:

in this example, the accuracy of the FGFR1, FGFR2, or FGFR3 fusion detection performance of the present invention was demonstrated by detecting 11 FGFR1, FGFR2, or FGFR3 fusion positive samples and 4 FGFR1, FGFR2, or FGFR3 fusion negative samples. The 11 positive samples included 10 positive samples (verified by F1 CDx) from beijing giegighur technologies ltd and 1 standard purchased from nanjing kobai (Cobioer), and the 3 negative samples included 2 samples (verified by F1 CDx) from beijing gieghur technologies ltd and 1 standard purchased from nanjing kobai (Cobioer). The initial amount of DNA library was 100 ng. The samples used were tissue and paraffin embedded tissue (FFPE) samples.

Firstly, preparing a DNA sample library of FFPE samples to be detected:

1. DNA extraction

The extraction of genomic DNA from the FFPE samples was carried out according to the QIAamp DNA Mini Kit extraction Kit instructions. Quantification was then performed using qubits, requiring tissue/FFPE genomic DNA to be greater than 50 ng.

2. Library construction

The FFPE genomic DNA was disrupted to 200-250bp, and then a sample library was constructed according to the NEBNext Ultra II library construction kit. Primers and linkers were from Biotechnology engineering (Shanghai) Inc.

2.1 end repair and addition of "A"

The end repair and addition "a" reactions were configured as follows:

components	Single reaction volume (μ L)
		End Prep Reaction Buffer	7
End Prep Enzyme Mix	3
		Fragmented DNA	50μL
Total volume	60μL

Fully shaking, mixing and centrifuging the mixture, and then incubating on a constant-temperature mixing instrument according to the following steps: firstly, incubating for 30min at 20 ℃; then incubated at 65 ℃ for 30 min. After incubation, the temperature was reduced to room temperature and the mixture was centrifuged briefly in a high speed centrifuge.

2.2 Joint connection

The linker ligation reaction Premix was configured as follows:

the amount of linker added varies with the initial amount of DNA, and the correspondence is shown in the following table:

31 μ L of linker-ligation reaction Premix and a linker of the corresponding volume were sequentially added to the reaction tube and ddH was used₂The volume of the O is supplemented to 95 mu L, and the mixture is fully shaken, mixed and centrifuged. Incubating for 15min at 20 ℃ by a constant temperature mixing machine. After incubation, the microcentrifuge centrifuged briefly.

After the ligation reaction was completed, the linker ligation product was purified using magnetic beads and finally redissolved in 25. mu.L of TE (pH 8.0).

2.3 introduction of index into Pre-Capture PCR (Non-C-PCR)

Reaction components were added to the PCR tube in the following order and a negative/positive control was set:

components	Single reaction volume (μ L)
		index Primer/i7 Primer-P7(10p)	2.5
index Primer/i7 Primer-P5(10p)	2.5
		2X KAPA HiFi HotStart Ready Mix	25
Adapter-Ligated library	20
		Total volume	50

Mix well with shaking and centrifuge.

2.3.1, the corresponding relation of PCR on-machine sample cycle number is shown in the following table

2.3.2 PCR computer program see Table below

PCR program for Illumina sequencer

PCR program for Gene + Seq2000 sequencer

The Non-C-PCR product was purified and finally dissolved in 31. mu.L of TE (pH 8.0). And carrying out Qubit-BR quantification and Labchip quality control on the purified product.

Secondly, preparing a DNA probe library aiming at FGFR1, FGFR2 or FGFR3 genes:

DNA probe libraries were prepared for introns 1, 4, 5 and 17 of the FGFR1 gene (1, 4, 17 in the performance report, and the detection range of 1021 is 1, 5, and 17, and 1, 4, 5, and 17 are included in the present application), introns 1 and 17 of the FGFR2 gene, and introns 15 and 17 of the FGFR3 gene (15, 17 in the performance report, and 17 in the detection range of 1021 is 17, and 15 and 17 are included in the present application).

In designing the probe, the overlapping region of the probe, the length of the probe, and the GC component are considered together. The probe overlap region is the number of overlapping bases between each probe. The larger the overlapping area is, the higher the coverage rate of the target area is; the smaller the overlap area, and even the spacing of probes, the lower the sequencing cost. The longer the probe length, the more tolerant to Single Nucleotide Polymorphisms (SNPs) when sequenced.

Finally, the probe length was determined to be 120bp, and the GC content was controlled to be approximately between 40% and 60%. Through screening, a total of 186 probes meet the standard, are synthesized by IDT DNA Technologies, and finally constitute a probe library for enriching FGFR1, FGFR2 or FGFR3 gene fragments.

And thirdly, enriching and sequencing on the target sequences aiming at FGFR1, FGFR2 or FGFR3 genes:

after the quality control of the amplified library is qualified, the enrichment probe designed by the invention is adopted, and the hybridization capture is carried out according to the instructions provided by the chip manufacturer. Finally, elution and back dissolution of 21. mu.L of ddH2O magnetic beads with hybridization elution are carried out.

1. Amplification of hybrid Capture products

1.1, removing the magnetic beads in the previous step, then purifying the magnetic beads, and finally dissolving back 25 mu L of ddH₂And O, performing QC and operation.

1.2, performing on-machine sequencing by adopting an Illumina sequencing platform or a Gene + Seq2000 sequencer, and performing on-machine sequencing according to the operation instruction provided by a manufacturer in the sequencing experiment operation. And computer data volume requirement: the FFPE sample subjected to capture sequencing by using the probe disclosed by the invention can obtain an effective depth of 500X.

Fourthly, information analysis of FGFR1, FGFR2 or FGFR3 gene fusion variation:

the method adopts an information analysis process (RealSeq Pipeline) of ER-seq (Enrichment & Rarallele Sequence) (Chinese patent publication No. CN105063208A, 11/18/2015) which is a low-frequency mutation Enrichment sequencing technology independently developed by Beijing Gioney plus science and technology limited company, and comprises the following specific steps:

1. based on sequence bases at two ends of an insert as a label, wherein the insert is a DNA fragment connected with an adapter primer in a library, and each fragment forms a pair of paired sequencing sequences through double-end sequencing; using the first 12bp base of the sequencing sequence 1 and the first 12bp base of the sequencing sequence 2 of the paired sequencing sequences as tags, connecting the smaller tags into a 24bp index in the alphabetical sequence arrangement, and using the 24bp as the index of the paired sequencing sequences, and marking the tag of the sequencing sequence 1 into a positive strand in the front; the tag of the sequencing sequence 2 is marked as the reverse strand in the front;

2. performing external sequencing on the indexes to achieve the aim of gathering all sequencing repeated sequencing sequences of the same DNA template;

3. performing center clustering on the clustered sequencing sequences with the same index, and integrating large clusters with the same index into a plurality of small clusters according to the Hamming distance between the sequences, wherein the Hamming distance between any two pairs of paired sequencing sequences in each small cluster is not more than 10, so as to achieve the purpose of distinguishing the sequencing sequences with the same index but from different DNA templates;

4. screening the repeated clusters of the same DNA template obtained in the step 3, and if the sequencing sequence number of the positive strand and the reverse strand reaches more than 2 pairs, carrying out subsequent analysis;

5. the clusters that satisfy the condition in 4 are error corrected and a pair of error-free new sequencing sequences is generated. For each sequencing base of the DNA template, if the consistency rate of a certain base type in the sequencing sequence of the positive strand reaches 80 percent and the consistency rate in the sequencing sequence of the reverse strand also reaches 80 percent, recording the base of the new sequencing sequence as the base type, or recording the base as N, thus obtaining a new sequencing sequence representing the original DNA template sequence;

6. re-aligning the new sequencing sequence to a genome by a BWA-MEM algorithm, and screening out the sequencing sequence with the alignment quality less than 30;

7. performing statistics according to the sequencing sequence obtained in the step 6 to obtain the base type distribution of each site in the capture area, and performing statistics on the coverage size, the average sequencing depth, the positive and negative strand interworking rate and the low-frequency mutation rate of the target area;

8. SV harvesting: according to the comparison of the information of the patient sample and the control sample, SV calling is carried out; the screening parameters used were: the variation rate of the control site is less than or equal to 2 percent, the number of variation sequencing sequences after error correction is more than or equal to 2, and the mutation prediction p is less than or equal to 0.05;

9. variant annotation: annotating the function of the variation, the number of variant sequencing supports, the frequency of variation, the amino acid variation, and the status of the variation in the database of existing variations.

Fifth, the detection result

The detection results of 14 fusion negative or positive samples by using the FGFR1, FGFR2 or FGFR3 gene probes of the invention are all consistent with the F1CDx detection result or the standard reference result, and the detailed detection conditions are shown in the following table, wherein the median fusion mutation frequency is 11.39% (4.07% -75.24%).

A DNA probe library for hybridization to FGFR1, FGFR2, or FGFR3 genes; the DNA probe comprises sequences shown in SEQ ID NO.1 to SEQ ID NO.186, and is shown in a sequence table.

While the invention has been described with respect to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention. Those skilled in the art can make various changes, modifications and equivalent arrangements, which are equivalent to the embodiments of the present invention, without departing from the spirit and scope of the present invention, and which may be made by utilizing the techniques disclosed above; meanwhile, any changes, modifications and variations of the above-described embodiments, which are equivalent to those of the technical spirit of the present invention, are within the scope of the technical solution of the present invention.

Sequence listing

<120> DNA probe library for hybridization with FGFR1, 2 or 3 gene and application

<160> 186

<170> SIPOSequenceListing 1.0

<210> 1

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 1

caccgccttc ccacaccctc ccagggccgg ctgcccgtga agtggatggc gcctgaggcc 60

ttgtttgacc gagtctacac tcaccagagt gacgtgtacg tgtcctgcag agctcaggct 120

<210> 2

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 2

ggtgtctgtc ctgggagtct caggacagcc tgacctcacc ttcccctgca gctggtcctt 60

tggggtcctg ctctgggaga tcttcacgct ggggggctcc ccgtaccccg gcatccctgt 120

<210> 3

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 3

tccaccgacg tgagtgctgg ctctggcctg gtgccacccg cctatgcccc tccccctgcc 60

gtccccggcc atcctgcccc ccagagtgct gaggtgtggg gcgggccttc tggggcacag 120

<210> 4

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

cctgggcaca gaggtggctg tgcgaagagg ggctcggtgg cacagcgctc accccgcctc 60

ccgccagcag gagtacctgg acctgtcggc gcctttcgag cagtactccc cgggtggcca 120

<210> 5

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

agagaagacg gaatcctccc ctgaggagca cgtagagctc cgggtgtcgg gaaagctggg 60

ggagtactgg tccaggggca tggacaggtc caggtactcc tgtgatgggc gagaggaagc 120

<210> 6

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

agcgatgggc cgggcccctc ctccctgctc agggaggtgc gtgcacgcag tggggacggc 60

ctgagctctg gctctggcac gggcagcctt acctggttgg aggtcaaggc cacgatgcgg 120

<210> 7

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

gctggtatgt gtggttgatg ctgccgtact cattctccac aatgcaggtg tagttgccct 60

tgtcagaggg caccacagag tccattatga tgctccaggt ggcataacgg acctgagggg 120

<210> 8

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

aaatgccaaa gggatacatt gagggtccag aggaaaatgc aggccccatg acaatgtcgg 60

caccccgtgg cacctgccct ccatatcaga gcctggtggc acagggcccc aggctgcagg 120

<210> 9

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 9

gttggctagg acaaggcgtg gattgccccc ctaccagccc gttcacactc tgcaagctgg 60

cagatggggt gtaaagactc cagaagtctc cactgtgacg ttcaagatca ttcgtgatcc 120

<210> 10

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 10

ggacagatgt gccttctgca aacactccca aatacaccaa gaatgttccc aactgtgcac 60

ctctcctatt acactaagaa tccagccccc actgtccttg acagctcatt tcaaaaacca 120

<210> 11

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 11

tctcctccag tccagtctct ggtcagaact gtgctcaccc tttcctctct gcctaccaaa 60

gcactgatca cagtctactt tttacggtag ctgctcatgg acatttgcct cctatcagac 120

<210> 12

<211> 119

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 12

tgcaaaattt ttatgggcag gaacgctatc ttgcagcatt taaagcatgg tgtcttgcat 60

gtagtgagtg catggtgaat taatttaaaa tatttctggc tgggtgcggt ggctcaagc 119

<210> 13

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 13

tgcatggtga attaatttaa aatatttctg gctgggtgcg gtggctcaag cctgtaatcc 60

cagctctttg ggaggcccag gcggacagat cacttgaggt catgagttca agaccagcct 120

<210> 14

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 14

ggccaacatg gggaaacccc gtctctacta aaaatacaaa aattagctgg gaatggtggc 60

tgcacatccc aggtactcgg gaggctgagg caggagaatg gcatgaacct gggaggtgga 120

<210> 15

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 15

ctgtcctaag gaagcacgct taaaaaaatg ctgacagaga ttcactgcaa cattatttat 60

tatggtgaaa aaccagaaaa tacctaatat cacaaaatag ttcattaaat tataggactt 120

<210> 16

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 16

tttctgttgt tttttggaga cagagtctca ctctgtcacc caggctagag tgaaatgacg 60

tgatctcagc tcactgcaac ctctgtctcc tgggttcaag caatctcctg cttcagcctc 120

<210> 17

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 17

cccagtagct gggattacag gagcctgcca ccacacatgg ctaatttttt gtatttttag 60

tacagaggag gtttcgccat gttgcccagg ctggtctcga actcctgagc tcaggcaatc 120

<210> 18

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 18

cgcccacctc ggcctcccaa agtgatagga ttttaggcgt gagccaccat gcccagccaa 60

attatgggat tttaaaataa tggaatctta tacagtcatg aaaaattatg tttcaaatat 120

<210> 19

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 19

tcatgcttga aatatatcag gaaaagactc atgttaacaa atgacaaggc aaggaaatta 60

catacatgat gtgatcaact atgtaaaaac catattacag aaagaagact gaagaaatgt 120

<210> 20

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 20

tgtgatcaac tatgtaaaaa ccatattaca gaaagaagac tgaagaaatg tttaaacttt 60

atttgcattt ttttgtattt aaaaatttta atgaataacc tgtataggtg gaaagtatta 120

<210> 21

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 21

cttaaaaaaa tgaaaagcat gtaatcagga cttcctaact cggcctcccc tgttcccatt 60

actctaactt tcgcatgcac acacacgtac cttgtagcct ccaattctgt ggtcaggttt 120

<210> 22

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 22

ggatgtccaa tatggagcta cggctgcccg gggaaagcca agagagacag gcagggtgga 60

gaggagcagc tggtcaggct caggagcagg gcccaggcca ggacctggag gtgtcttgcc 120

<210> 23

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 23

gaaacagtgt ctcacgcata cggtttggtt tggtgttatc tgtttctttc tcctctgaag 60

aggagtcatc atcatcatca tcatcctccg aggaggggag agcatctatg ggaagaagaa 120

<210> 24

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 24

tccatggata ctccacagtg agctcgatcc tccttttcaa actgaccctg aggaaaggaa 60

aaaaacccca aaagttagga gggtctaggt aggggagggg aaaggaaaaa cagaaggtaa 120

<210> 25

<211> 119

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 25

agtatgccat gtggtggctg cttaaagtgt ggacttacta aggcgtccag aagaaaattg 60

tgaagctacc ccaacaatga attagatcat aggtcctggc tgggtgtggt ggtgcaccc 119

<210> 26

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 26

cagcctggcc atggcgaaac cccgtctcta ctaaaaatac aaaaattacc caggcgtggt 60

ggcgcgcacc tgtagtccca gttacttcgg aggctaaggc aagagaatcg cttgaatcca 120

<210> 27

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 27

ggaggcgaag gatgcagtga gctgagattg caccactgca ctccaacctg ggtgacagag 60

cgagattctg tctcaaaaag agaggttata ggtcttaatt ggacagaggg atgctttagt 120

<210> 28

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 28

gattctgtct caaaaagaga ggttataggt cttaattgga cagagggatg ctttagttca 60

attctacagc tatgttttga gtgcagatta cgtgcaagcc tgggaatacc atcatgaggt 120

<210> 29

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 29

caagcctggg aataccatca tgaggttttc caaggtataa acttcaaagt agtatgtgct 60

acaaagtgaa tggtctcatg aaagtcccaa atagttctgc ccttttgtgg gaaacagttt 120

<210> 30

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 30

ttctgccctt ttgtgggaaa cagttttaca aagtgtgcta caattgcagg aggtttgagc 60

atgctgatgg ctccccagca taagacttca ctctgcccag gccaggtgcg gtggctcaca 120

<210> 31

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 31

ctccccagca taagacttca ctctgcccag gccaggtgcg gtggctcaca cctgcaatcc 60

cagcacttcg ggaggctaag gtgggcagat catttgaggt caggagttca agaccaacct 120

<210> 32

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 32

ggccaatatg gtgaaatccc atctctacta aaaatacaaa aattagctgg gcatagtggt 60

acacacctgt aatcccagct actcaggagg ctgagacagg agaattgctt gaactcagga 120

<210> 33

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 33

ggcggaggtt gcagtgagcc gagatcgcag cactgcacta cagcctaggt gacagagtga 60

gactctgtct caaaaaattt aaaaaaaaaa ttttaaaaag accactccat ctagggccaa 120

<210> 34

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 34

caaaaaattt aaaaaaaaaa ttttaaaaag accactccat ctagggccaa ccagtccctc 60

tggtgagaaa taaggggcaa tgaagaatca cggaacaagc taaggaccag taaatggaag 120

<210> 35

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 35

aaatggaagc caggtacaga gataactagt tagtagctgc tggactccat gctcaactcc 60

ccgacctatg caatgaagca gctgaaataa atgacctcct gggtactttc tttttgtctc 120

<210> 36

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 36

ttttgtctct aaaattgtac gaatcacatc catagttcta ccagcctccc ttgcagagag 60

aagaaaaccc tcaccagcta cttacaaaga gcctaagagt gtggagccag gagggatcca 120

<210> 37

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 37

agggatccag ttctccctga tgctggaagg tgacaagtga tgtctgacgg atatgagtcc 60

agaagttgcg gggggcaagg gggcctggga ggtcccactg cagaatcaag gtctattgca 120

<210> 38

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 38

tctattgcaa gcactgagtc aaagatggct cacactggcc ccagttaaaa gaacagaatc 60

tgcagccctt tacccttctc aggtctggtg gggtaaaact taaaactccc cagggtctta 120

<210> 39

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 39

agggtcttaa agaacggatc atagctgtta acatttacca agtgacaaga ctttctgggc 60

actgtcaagg ctacgtgggg gtgaccatct ctattttcca ggtggcttct gtcctaaatg 120

<210> 40

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 40

tcctaaatgc tccagggagc catccaagtt attcagagaa tctacttccc cggaacatgc 60

cctaatgtgg tacttcagcc ctgcctcccc cgagccaggc agggcccctc gcaagtgagt 120

<210> 41

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 41

caagtgagtc agtgctggct ggcagaaaag cccctggatt cttggcttcg ttcacatgtg 60

cttggccagc gggatacaca tgccacacca ctagctccct cccaaggctt actaaaatgt 120

<210> 42

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 42

ctaaaatgtt ttaatgattt caaatcaaaa gagaagtcaa catttctctt tcatttttct 60

acttttacga aaggaatcag aaaagagaaa atgaaatgaa tactgaacac ccactatgta 120

<210> 43

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 43

tcagaaaaga gaaaatgaaa tgaatactga acacccacta tgtaccaagg atgccctaag 60

tagaattaat ttagaactat agtgaggaat taattcattc ctcactataa acttatgggg 120

<210> 44

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 44

gataagtttt attattcctt ttatgcaatg gagaaactga ggctcaaaga ggttaagtac 60

tttactcaag gaaatgtagc tagcatgtaa tagtgcctca aggtttttgg atattagaga 120

<210> 45

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 45

agctagcatg taatagtgcc tcaaggtttt tggatattag agaacagggt gggaggaagt 60

aaataaggga ttgatcaagc ctgactccat ctgatgcaaa ctcaatttat ccaaaccaag 120

<210> 46

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 46

tatccaaacc aagtagataa gggtaaatcc aaggtaactc tggtaacaaa agaggcttga 60

tttgcagttc cattggtcaa agatagaccc ttgtgccagg gcatgttggc tcatacctac 120

<210> 47

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 47

aatcccggga ctttgggagg ctgaggcggg agtatcacct gaggtgggga gttgagacca 60

gactggccaa catggtgaaa ttcctcgtct actaaaaata caaaaattag ctgggtgtgg 120

<210> 48

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 48

tggtgcacgc ctgtaatccc agctactcag gaggctaacg catgagaatt gcttgaattt 60

gggaggtgga ggttgcagtg agccgagatt gcatcactgc actccagcct gggtgacgga 120

<210> 49

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 49

agtgagactc tgtctcaaaa ctaactaact aaaagaaaga cccttggaag cagaccacag 60

atgggccatc taggacactg catggggcac attcccagag ctgggataga agcagtgctt 120

<210> 50

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 50

attcccagag ctgggataga agcagtgctt gccagaggat gcccctaaac ccatattcca 60

caacttacta catcccgaaa atacacctga tgcaggcctg tcacccagaa acgcacacct 120

<210> 51

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 51

tgcaggcctg tcacccagaa acgcacacct gaccaagaaa acccaggtcc cttgctaagt 60

gtcagaatac tgaacaagtt ctcaggttcc aatgggaaag agtggtgtca tgttgagatg 120

<210> 52

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 52

aatgggaaag agtggtgtca tgttgagatg gttaagcttg cttcgacttg gccactctgt 60

ccccagaaaa gcctgccgcc tcctcccagg tgtgggctgg ctgcagaccg ccctctctca 120

<210> 53

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 53

tgtgggctgg ctgcagaccg ccctctctca gctccctggt caatgcactt caaattcatc 60

tgccagcttc cctcttgttc tttcccaaaa aaggtacaaa aagtctcttc ctctcctacc 120

<210> 54

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 54

tttcccaaaa aaggtacaaa aagtctcttc ctctcctacc cctctggccc tcaagtttga 60

catctgtaaa acaaggatgt taagaaaacc tggcctcctg gcaccaactc ctggaataag 120

<210> 55

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 55

ttaagaaaac ctggcctcct ggcaccaact cctggaataa gaagtcagca tacaccccat 60

gccacccgcc tgcaaaaaac acagtgacac aggccactag tccaacatgc tgtctgggac 120

<210> 56

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 56

gtctgggacc ctgccacctc ctgcttcacc tcagtttctc attctactaa ttaatgaatt 60

tttccaatcc cttagaaaca acttccttcc ttcactgctt gacagataga tgtaacctgc 120

<210> 57

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 57

gtaacctgca gccttgcagg tgtacacagc agcttcctcc aagaccctgc agctggttcc 60

ccccgccccg tctctccaat atcagctagg ctcctgggat tcatgcctgc atgcagaggt 120

<210> 58

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 58

tgcagaggtc ctcggtgggg agcagagagg gggcacatct cagtttccca ctcatcagag 60

ggggctgaaa tacttgcctt gcctccatca caggtggtta tgaagactga atgggctgca 120

<210> 59

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 59

tgggctgcat gggtgaaagt gcctgctgtc ttctcactgg agtactgatc caacatacag 60

ggtggacagt gtctggtgag atagcagggg ctggacggac cacgtgacct tgaagctctc 120

<210> 60

<211> 118

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 60

gaagctctcc cacttctaag agaaaaacct atgaatgtca gatctccttg tccaaaaatt 60

atgtaaggaa aactagctga cgaattcatg gccaaatctt tttattcttt ctgagaca 118

<210> 61

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 61

tttattcttt ctgagacaga gtgttgctct gtcacccaag ctagagtgca gtggcacgat 60

ctcaactcac tacaacctcc atcccccaga ctcaagtgat cctcccacct cagcctcccg 120

<210> 62

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 62

agaagctggg ctaattttct gtatatttaa taaaaatggg gtctcaccat gttgcccatg 60

ctggtaatga actcctgggc tcaggagatc cacctacagt ggcctcccaa agtgctggga 120

<210> 63

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 63

ttacaggcat gagccaccac acccagcctg ggtcttttta ttcttgtccg ctggagacag 60

tatgaattca cacccttcca cttatgatct tgccaccctg ggatagtcac ttaaccgttc 120

<210> 64

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 64

taagcctcgg tttcccacct agaggaagcc acatcacagg tgattatgag gctcaaatgg 60

gtggtgtggg tgactgtgcc cattctctcc tctctaaagt accaaccaaa tgtaaagaat 120

<210> 65

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 65

aaccaaatgt aaagaattat gcatttggat ttcagatata atattaagat atttataatt 60

ctctttgtgt agtgctttga gtcactcctt aatagcactc tcaggagatt tggtaattgg 120

<210> 66

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 66

gagatttggt aattggcagc agaaggacag gaacttccaa cctttccaaa ggcatttctc 60

actctccatc ttcatgtctt atattctaga cctgtggccc acacagtggc cactgcacac 120

<210> 67

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 67

acacttgaaa tgtggctggt gctaacggat atgctataag taaaaagtat acaccgcatc 60

ttaaaaactt gttatgaaaa aaatgtaaag tatctcatca ataatttcta tattgattac 120

<210> 68

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 68

atgttcaaat gataatattt tggaaatata gttacattaa ataaaatcta aacaaacaag 60

ttaattaaaa aacaaatatt ggccaggcgc ggtggctcac gcctgtaatc ccagcacttt 120

<210> 69

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 69

gggaggctga ggtgggcaaa tcacgaggtc aagaggtcga caccatcctg gccaacatgg 60

tgaaaccccg tctctactaa aagtacaaaa attagctgcg tgcagtggcg cgcgcctgta 120

<210> 70

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 70

aaaaaaaaaa ttttttttac ctatttcttt cttttaactt aatttccaat gtggcttcta 60

gaaaatctga acatatacat aactcatgtt ataatgttat atttctttct tcctttcttt 120

<210> 71

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 71

ttttctttga gatggagtct tgctctgtca ctcaggctgg agttgcagtg gcgcaatctt 60

ggctcaacca ccgcctcccg ggttcaaacg attctcctgc ctcccaagta gctgggatta 120

<210> 72

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 72

aaagggctgg gattacaggc atgagccact gtgcctggct tttttttttt tttttttttt 60

aaacagggtc ttcctctgtc ccccaggctg gaatgcagtg gtgcaatcac agcttattgc 120

<210> 73

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 73

agcctctgcc tcctaggctc aagtgatcct cccacctcag cctcctgaat agccgggact 60

accagtaagc acgagcatac tcatctaatt tgtgtatttt ttggtagaca cagggtctca 120

<210> 74

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 74

ccatgttgtc caggctcatc tccatctcat gggctcaagc aatccgcctg ccttggcctc 60

ccaaagtgct gggattacag gcatgagcca ctgtgaccag ctgctcatgt tctatttcta 120

<210> 75

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 75

tagggcgcca cagctcgaag caacatccat gcctctgtgc aagactttct tcatgccatt 60

ccttgttctg caacaggcca gcccttccct tttctctgaa ccccacctta agaagctgtt 120

<210> 76

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 76

tcttccataa gactttcttg ggccttctag ctggagaatt tcattcctcc aaatttgcaa 60

accacattac tagcccctct cacatagcac ttaggtcata ttatatgatt gttatttatg 120

<210> 77

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 77

ttcttcattc tcttctctcc ccacgaaact gcaggctcct tcagaacagg gtctgtgcct 60

ccctaacctt tgctaacacc ctcggcacct agcatgtaca caggagatgc caacatgttt 120

<210> 78

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 78

tgtttgttaa ctggatgaat gagtaagtga atgaaggaaa aaaaagggct gatgaaaaga 60

aactgacaga gatactcctc cagaaaaaga ggctgagaat atcagagaaa agcaattagg 120

<210> 79

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 79

gtagtacccc ttgttaacca gccaaactct agtccaaaga accctgagaa aatttctgga 60

caacttggtt cttcagaatt gaggaagtaa caccaaaaca ttaatccctt cagatgagaa 120

<210> 80

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 80

ataagactta tttcttcccc aaagtgacca ttctctactc acatgttgag agggcagggg 60

gcaagtactt aataaacata tggagaaagt tactcagctc agcttgctaa attttagttg 120

<210> 81

<211> 119

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 81

ggtccaatgg aaatgctaat tttttgagtc tctttaaaaa aaaaaagtgt ctaaatgtcc 60

ttggaagtaa gagacatata aaaaattagc aataagtggt caggcacaat ggctcacac 119

<210> 82

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 82

agagacatat aaaaaattag caataagtgg tcaggcacaa tggctcacac ctgtaatccc 60

agcattttgg gaggccgagg ccagcggatt gcttgaggtc aggagttcaa gcccagcctg 120

<210> 83

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 83

gccaacatgg tgaaacacca tctctattaa aaacacaaaa aattagccgg acatggtggc 60

gcatgcttgc aatcccagct actagggagg ctggggcagg agaatcgctt caaccggaag 120

<210> 84

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 84

gcggaggttg cagtgacctg agatagcgcc acagcactcc agcctgggcg acagagcgag 60

actatctcaa aaaaaaaaaa aaaaaaaaaa agcaacaagt gaaacacgaa ctgctgcaca 120

<210> 85

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 85

aaaaaaaaaa aaaaaaaaaa gcaacaagtg aaacacgaac tgctgcacat aatctagaaa 60

accttgctct tgaagacagg atgtactatg agtggaaccc caccaaattc ttgggatagt 120

<210> 86

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 86

tcatccatgt gagggctatt aaatggggga gagggcccta aacatggttt ctttcattgc 60

aggggagaga taaatggtgg cgaggagaga atttctgttt aaagtagatg caaccttagg 120

<210> 87

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 87

aggaaaaaca aatatcctag cccaagtctg acatttcatc ctctctggaa acaccagaat 60

cttctctaaa gctctgcatc tgggctccca ctagtccctg gtgcgttcag gccttttaca 120

<210> 88

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 88

aagctctggg gaagacaaat gtaagacaca aatttcccag gaggaaatta aactatcaag 60

tgatgtgagg ccctttcaga ataagacatt tgaatcaatt ctaactcctt ttcctccagc 120

<210> 89

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 89

agcaacaggc tcaaactttg ctaggatgtt ctataactaa cagccatttt gacgaaagac 60

actttagaaa cgaaggcaag aaaaaagcct ttgtagaagc cggctgctaa acccccaagt 120

<210> 90

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 90

tcataccagc aaatctctca ttttaggagt cttgtttcag gtcaatactg aaaacaaaga 60

gcacaccact aaaagcaaaa cccatggatt ttccttggtc tctaaggctt gaatctgcct 120

<210> 91

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 91

acccagtgtt caattcaatt ttcttgccct ctggagtcca ccaggaataa agctagccca 60

gcctgaggga tgcaccgatg tatctaatcc tctgccccta gggagtgggg gacgcttctc 120

<210> 92

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 92

catttcttcc tgagcctttg cttcccttgc tatcaaaggg ccacctaatg gaactgtcta 60

acaccctcca tggccagagg ccaaggaaaa gaagcaccct aagattgttc aagtggtgcc 120

<210> 93

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 93

cactctggcc tgaattccca ttgggaaaaa tacctcgaaa atgagcattt ctgcaggcat 60

gtttggaaaa gcaggcagtg aagtacgcag tccaagtggc tctcaatctg gatgaacctc 120

<210> 94

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 94

attaaaaagc tgacactcag ctatgactaa gagtaaatgc aacagatatt tcaagcagct 60

cgtcactgat gcagttcata accctcccag ctggagtcag gccccagcag cctcccggtc 120

<210> 95

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 95

cccaaacacc ctgcactcgg agggtcgggg cagccccgga gcttctccta acccactggg 60

ggtaggatat gtaaacaagt cattagggag gcaggccatc ggcaaagacg gccggcaata 120

<210> 96

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 96

gcagaggctg ctgaacctcg gtcaccgaga ggaaggtgga tgacccgggc gtgggggtga 60

ttacccagac caaggcgcgt cccacggggc cctgctactt cgcttttaaa caagcctcag 120

<210> 97

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 97

ggaggaaacc gggagcccgg gccaagggag gaaaatggcc actttctgct aaacagtctc 60

aaggctagat agtattccaa aaaaagcgct ggtccggagg tgcaaagggt agggcacatc 120

<210> 98

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 98

tgcggaggaa acagtcacct taattaccac gtctagagaa agtgcgattt cattacagcc 60

tccaaaacta ccgcccaaag aaggaaacca gagaagtgtt ttcaatttga aagaggctgg 120

<210> 99

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 99

aaagaagcgg gtaagacacc gactctcccc accctcccgg caactttgga ccacaaggcg 60

ctttcctgcc tcaaagacta ttaccaaaca caactcgaaa atccaacccc gctttcaaac 120

<210> 100

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 100

ctgtagccgt tatttaaacg gtaaaacacc ccactttagt ttccggggga tgcggcccaa 60

atcagtcctt aaaaacacac agagagcgcc tctggcagca gccgcccacg cccccggccc 120

<210> 101

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 101

tgcgcaccgg gcttgctccg ggaggggcgc gtgacgccac cgcggggcag ccgagagggg 60

accgcgggct ccgggcaggg caggcgctcc cgcagctcca gctataacac aagggagaga 120

<210> 102

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 102

aagcaacgct ttggttccag aaacttccat cgtcaaggcc agttctggcc agacgcacag 60

ccaaagagcc cccctagtct agcggatgaa ccgttcgcga gtggcacccc ctaaaacctt 120

<210> 103

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 103

ggccccatct cccaagatgc ctctttactg cgcaatccag aaagcacccg gaccgccgcc 60

acccaccagc ggcggcgaat aacttggtga gatcttaaat aactcgggat ccaagggaac 120

<210> 104

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 104

tttcttttct cccccgaatc tcaacgccgc atctccaagt ctccagaaat gttaaggagg 60

gaggtggtca agacaggctg aagaatctga gttttaattt gggggagggg gaggggagga 120

<210> 105

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 105

agagggaagt taataatgcg gctgcggtgt acccgtgtgg ggtgaaacgg aaccagatgc 60

ggctgctaca tctggaaatt gggggtgggg gagtgctggg ggagaaaggc accgtggtaa 120

<210> 106

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 106

tgtttttcaa aagggttgat agcttctggg ctcagaacaa gaaacaggct tcacacccca 60

cccccccccc accaccacca aaaaagtgtc tggctgcctc atagggggtg tgtgttctat 120

<210> 107

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 107

atgaaccccc atctacaagg tccataaaat tcctccgcaa acatgggtcg tagcctcact 60

tttctcccaa ccccaaacat aacggtaggg gaagcgaggt gcagaggggt ctgggccaga 120

<210> 108

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 108

gaagcatacg catcgctgcc gagtgaacca caccggacaa aaaccctcat cccttccccc 60

acctctagga accagattgg gaccgaacgg tccttccgcc cccatcccac acctcggctt 120

<210> 109

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 109

aacgaccgtg ggttgtcctc ccgttcccca aacttccgac cctttcagag accgtccaag 60

tcataagatc accccttctg gaaaggggag tcgtttttcg ctctcagctc aagcagattc 120

<210> 110

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 110

agccatttgg tgacgacacc tccttaaaac ctattccaag agaagctaac ctagccctcc 60

acgtcctccg cagttcatct cccaccccat ccgaccccaa cccgcacccc aggctttccc 120

<210> 111

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 111

gggggtggaa aaggacttag gaatgccccg gatttttagc gttgagccca aagctacctg 60

ctgaattggg ggtgaatccc caaaagggga ccaaaaagac atggagtgga atgcaacaca 120

<210> 112

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 112

cacacataac acacacaacg catcccacct cgccgcccac tcccgccctc ctccccagtc 60

caggaaaccg cgggcctcgc cgccttcgcc ccgggcctgt ccctcccggg actcactgag 120

<210> 113

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 113

gagccgccgc cgcctcgcca gctcccgagc gcgagttgga ggaaaagttg ggcggcggga 60

agagcggcgg gacgagcgca gggagggggc gcaggagacg cggacggagg gaagggaggg 120

<210> 114

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 114

gagacccaag gggcgcggat cgcccgggag ggagccagga ggtgaaaggg gcgggcggcg 60

agcggaggga ggcgccggcc ccggctgggc tgcggcgggc aaagcgcgca gccgggaggc 120

<210> 115

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 115

tccggcgccg ggggccgctc gggacgccaa gcccgcccca gatccggggg cgccgcggct 60

cttacctcgc tcggcgtgga ggttccgcct cgggagagtc cgccgtggct tgtgcgagcg 120

<210> 116

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 116

ggtgaatact gttcgagagg ttggctgagg tccaagtatt cctgaaagaa gggaagagag 60

acgttttatt tcatcttggg tcaggataac aaggtgaata cggttcgaga ggctgactga 120

<210> 117

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 117

ggtccaagta ttcctgaaag aagggaagag aagagtttta tttcaaacac aaactcttga 60

gatgtgggta ttggacgtgg gacaggacag gagaccccta gaaggtgaac agagacccag 120

<210> 118

<211> 119

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 118

cccacctgac ttccacgaag acctccctga gaccacgtct gatgtacccc aggctgtcct 60

tagcttttct cttggaatcc atcataaatc aactattgac aggtagagaa caataaaac 119

<210> 119

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 119

ttggaatcca tcataaatca actattgaca ggtagagaac aataaaacct agttgagcta 60

taggtgctgg gcaggcaact cggcttcaca cggacctaaa taacccgctg agtctacacc 120

<210> 120

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 120

acctcctcac tagagccctt gcttcaaatg cgagccccag caaaatgaaa accggtaaca 60

tgttgtgggt ccccagaccc cctttaagta gttgagacta tgaatattta ttttctgaat 120

<210> 121

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 121

tacattttac tattgtagtt tcggggataa ttgaagggga atacttcttc attcagtggg 60

taatttaaca gatgtaactg attactctca gaagctctcc gagttgaaaa tgtaaacagg 120

<210> 122

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 122

ttcaaaaaag gcttagacat actcatgtat gttaaatcca taatgcactg ttaaaggaaa 60

cttcaggaaa atatcggtta agtttcaaag ttttgaaggt tcgagattca gaaagtgtcc 120

<210> 123

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 123

atcggttaag tttcaaagtt ttgaaggttc gagattcaga aagtgtcctt tgctccccaa 60

ggcatccagt gctggtgctg caaaaagccc tctggatcac cccagggtga cacttcctcc 120

<210> 124

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 124

gtcctttgct ccccaaggca tccagtgctg gtgctgcaaa aagccctctg gatcacccca 60

gggtgacact tcctccgact tgggtgccca cagacgactt tgtatggcct gagggctaaa 120

<210> 125

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 125

ttcctccgac ttgggtgccc acagacgact ttgtatggcc tgagggctaa agatggtctt 60

taacattggt aagcggttga aaaaaaaaat caaaagaaga agaatatctt ataggattaa 120

<210> 126

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 126

aatgaataaa atttatatac aattatataa aattccaatt tcaggatcca taaaatttta 60

ctagaagaca gactagaaca catctattta catattgtct gtggctactt tgaggctaca 120

<210> 127

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 127

gtggcgaagt tgcaacagag accacagggc cggcaaagct gaaaatattt actccctggc 60

cctttacaga aagcagttgc cgacccctgt tctgaagcta tgaaaagctt tctaaattgg 120

<210> 128

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 128

aaagcagttg ccgacccctg ttctgaagct atgaaaagct ttctaaattg ggaacttgtg 60

ggtctctctg cctccactgg gctggcacag tggggcagtt ctgctcacac atccagaagc 120

<210> 129

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 129

aagccaacaa ctattctatg acatgtggtt ttgctgagag ggggcaaaaa gtgcagacca 60

cagcacaaac agcctcagtg ttatcagaaa agagcccatg gaatgttggc aactagacat 120

<210> 130

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 130

acatttttaa agacacacac acacacacac acacacacac acacaaaacc ctctaattca 60

tcaccagggt tggcaagaga aagagtcccc atcccagcat cccagaaggg gaatgtatca 120

<210> 131

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 131

atcagttcca acaacatcag cattaaccac acaactggaa acaaacacgc ccaccacatc 60

aaaagagaac tttttgctta caatgataaa aacgaaattt tgtcctaaat ggaaccgttt 120

<210> 132

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 132

gtttttcttg agcatatggt aatgattttc agaaggaaag aaacttcgat ttttatatcc 60

accagacaat atggcattct gcttttttat tttagggtac tagatacaaa tttcagtatc 120

<210> 133

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 133

tatcaacgga cccactctac atgtaaatat ataaataaat cactttgagc atagacatga 60

gatcattaaa atagaacttt agtgcttttc ccggtttctt tcttttttat ttttattttt 120

<210> 134

<211> 119

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 134

tcaacggacc cactctacat gtaaatatat aaataaatca ctttgagcat agacatgaga 60

tcattaaaat agaactttag tgcttttccc ggtttctttc ttttttattt ttatttttt 119

<210> 135

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 135

ctttagtgct tttcccggtt tctttctttt ttatttttat tttttttttt tttgagacgg 60

agtctcgctc tgtcgccaag gctggaatgc aatggcatga tctcggctca ctgcaacctc 120

<210> 136

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 136

cgcctccccg gttcaagtga ctctcctgcc tcagcctccc ggagtagctg ggattacagg 60

tgcacaccaa cacgcccggc taattttttg tatctttagt agagacaggg tttcaccatg 120

<210> 137

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 137

atgagccact gcgcctgacc aacttttccc agtttctcaa tgaagccata aactttcaga 60

tctgatagga aaaaaacagg gatatcagta gattccaagt ctacagttaa aggaacttat 120

<210> 138

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 138

atactagaaa caacaatttt ggcagaagaa gaaagttggt ttcttccccc ccttgaaccg 60

ttcctttcct ttcaacttct gaaggtcagc tgtattcatt taagtgcaat ctcttccttt 120

<210> 139

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 139

ttatccagca caggccacag tcagacgcat aagataaagc tgctgagcta agacttcata 60

tcactacatt tgattattac tttttaaaaa atcactctaa aactatttcc taaatcagaa 120

<210> 140

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 140

aaattaagtg tattattttc cttctaaaat acttggggga ctttaacttt tttcttgttg 60

agacaattgc atggtttgga gttagtttcc tgtgtaaaag aacactattg tgcatcctca 120

<210> 141

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 141

cattaaatat ctcaaaatac attcattaac aatttctcta tcttaagatt tgactagact 60

tgatgggttt atcatctttc ctgctcagag catatgcttt ccataattat aaaaggcact 120

<210> 142

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 142

atgctttcca taattataaa aggcactcta tctgcatatt gaaggagcca ctgacatggc 60

taaatttgca cagtgagtga cagacgagtg tttattttgt tggcaaggat cataaattat 120

<210> 143

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 143

ctctatctgc atattgaagg agccactgac atggctaaat ttgcacagtg agtgacagac 60

gagtgtttat tttgttggca aggatcataa attattatta ttttcttttt gagatgaagt 120

<210> 144

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 144

gtcttgctct gtctcccaga ctggagtgca gtggcgcaat ctcagtcact gcaaacttca 60

cctcctgggt tcaagcgatt cccctacctc gcctcctaag tagctgggat tacaggcaca 120

<210> 145

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 145

taccaccaca tccaactaat ttttgtattt ttagtagaga cagggtttcg ccatgttggc 60

caggctgctc tcgaactcct ggcctcaggt gatctgccct cctcggcttc ccaaaatgct 120

<210> 146

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 146

ctgggattac aggcgagagc ataaatattt tagggaaaat aaatatttta acaaaaggta 60

catgttttta tgtaaaaatc atgtaagttg ctcattgtag aaaacagtgt atgtttatat 120

<210> 147

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 147

acacctagaa agaggctgca aggcatatag cattaccaac aggatcacat aggatagtac 60

tgtggtaaca gaagctggtg ttagaaaact tcattgtatt ccatcaagca aatgatgacg 120

<210> 148

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 148

aacattatca gtttacccac taatgaccaa agcacagaat caaacttgca caggaatcca 60

agctttttcc catgcacacc aaaatgcaag tgaatactgc ttcccctttc ccactctccc 120

<210> 149

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 149

acataccccg gactgctcca ccagcagcta cactgaaggc agcctcgggg atgtccctct 60

gtctttggtt tcatgagtca gaactcacaa aatgaagttc tgagaaaaat catgatttga 120

<210> 150

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 150

gaggaaccca tagttgaact atctgaccct atggaaaaag aataaacaag accacagact 60

ccaaccaaca gccaacaggg gagtgtgtgt gtaaaacact acgcatgtct cacaagacaa 120

<210> 151

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 151

ccaaggacaa ggggcttcta gaaggaagtt cttacctcat tggttgtgag agtgagaatt 60

cgatccaagt cttctaccaa ctgcttgaac gttggtctct gggagggcac tgcatgccaa 120

<210> 152

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 152

ggtgggctca ggaaccgagg cgctgccgct gctgctgcag tcactaaagg aaagagattg 60

gcgagtcagg gaatcttccc caatgccaaa tcactccagc ccaagtggga taaaaccatt 120

<210> 153

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 153

tttcagcctc tcaaatgtgc gcaaacagag ttctttttca ttatctgcta aatccttcca 60

tcctcacctc agagtcccag gtgataccac cagaccggtc cacagaaatg tgtgaggcta 120

<210> 154

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 154

atctttttaa aagcatactt ttaagaaaca gagataaaac agtagctaca ggactcagat 60

acgtgcagcc acttcagtga ggcaatgtgt gctctcttct actatccact atcgcctcca 120

<210> 155

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 155

ccgccctcaa tgagcagaag caacgtgacc ttagtttcaa gggcaactac aaaagcaagt 60

taaaaatgct cgggttggca taaccaacct cgttcccttc tcttagtcca actgggcttt 120

<210> 156

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 156

agaaagctaa actgttggag gagtctgctc acagctccgt atttctgctg ggctggaggt 60

gagcagtctc ccaccctcac tgtcgctctt gggaagaaag gagaccagca aaatcaaaag 120

<210> 157

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 157

aatcccagga aggctacaga aaattcccac cagtttacac ttgtcaggat tcattttcaa 60

atggagaagt gtctttcagt cttccgccaa gcagccactt acgaagagtg aacacaactg 120

<210> 158

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 158

cagcgcattc aacgcagctg atccaaaaag gacttcgaag cctgaattcc cttcccctgt 60

gactaaatgg cagtccaggg ctgctaaatg tcttcaacct ctgtagaagg gtgaggaaac 120

<210> 159

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 159

tgagcagatg caaattctga tgaataatgc aggaatttat taataatgat gatgtaaatt 60

atgtaaatta tagtcatggt atgtgattgg accaataact ttccatatgc aaacctactg 120

<210> 160

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 160

accaataact ttccatatgc aaacctactg gctaaatttt cagagtataa gttagaagag 60

gacactcaac ttaatattct gttcacccat ttgatccaag ggaaagagaa ttttccaaga 120

<210> 161

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 161

gctaaatttt cagagtataa gttagaagag gacactcaac ttaatattct gttcacccat 60

ttgatccaag ggaaagagaa ttttccaaga aagcttaata tattccaagc aactttaaat 120

<210> 162

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 162

tcactcagta ctctctgtga ctaagggtac agtaaaatgc acaaacttta gagaatgatt 60

aaaacaatca caagaagggg ctcatcaatc tcaggcttca gattctgtga accacaggaa 120

<210> 163

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 163

tgaaccacat tttctaagac tgggggtggc agggaggcct ttaatacaaa aatctctaaa 60

ctctacattt tttatgtacc cactcgatgt ttcaagttat gtcataggat ttcaacactg 120

<210> 164

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 164

ctggagcctc cccaaatctc atcataaaaa ctgctccttt tagctttgac tgaggacaaa 60

aatggaggag ggaaaagaat atgccacatc atttccaaag gaggaataaa gaaaacctct 120

<210> 165

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 165

gcttcaaaga tagcacaggt gaaacggtgt gattcacagc aaaagcaaga aaagtatgtg 60

tgcagtgtgt gtgtgtgtac acggtgtgtg tgcgtgcatg gtgtgtgtgt atgcacggtg 120

<210> 166

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 166

tgtgtgtgta catggtgtgt gcgtgcacgg tgtgtgcatg tgctttcaga tagcttgaaa 60

gatttttcct cttaactttt ctttcctctt agtcaagagc ctcagaagat aatttcggat 120

<210> 167

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 167

ctatttctac atacatgtgg gaggaaagta cacaaacatt ctcattgatt tactaagcat 60

ttgcagaatg caggggaatt ttctacctct aacacgactt caggaacaat cctaagtcct 120

<210> 168

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 168

cgcagaggtt tcaccccagg ctccttcccc ggcctgctcg cccacctgtc cctgcccaga 60

ggagcctcct tggctccagg cagagaaagg tggacagggc gctccgctga atctcaagcc 120

<210> 169

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 169

gttccatcat cccgctagct ggcactagaa atttgcattt cagccatttc caggtccaaa 60

ccccacggca gccccctccc caaaactctg gaaggcggag acctaaggcg acaggtctgc 120

<210> 170

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 170

acagttcctc acaccagtgg acaaaagtat ggtgcgacct cctaggcagg ataattaggt 60

ccccaggctc ggtgttggga atacaggccc agcatgccgc tggcggccac aatcccggct 120

<210> 171

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 171

tcctgccttc aataccataa tccttcctgg agtctgagcc ccagtgaaag ccccagcgtg 60

tttatttttc cccttgaaag ggaactctct tctcagcctg cacgcgggag gagttgtcac 120

<210> 172

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 172

cctccctctg atggatggct cttctcggcc gcccgccccc cggtccattt tcacctcggc 60

acaagcctgc ccccaaagac aggcagagtg acaagcagga agggcttggc tgcggagcgg 120

<210> 173

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 173

tccccatgcc aggcctgtgc tgggctcttc aatttaccat gacattccca aaacccccgc 60

cctccaagaa ggcagaccaa tacagtgccc tccgctttgg gcttccacct aaggggtgat 120

<210> 174

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 174

ctgtccactg ctggctcggc cactgggatc acctggtgtt ttgttcctgg gtagatttca 60

catcaggaaa gtcaacagtg tccggctcct ccaaagtgca gacccatctc ttcgcttaag 120

<210> 175

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 175

gcctccctac ttctccccag cactgagtag gagcgggagc tgtcaagggg tcaaggctag 60

ggggaaattc ctcaacaaat tgaaatctca aaaaacacac actgacccag gaccacaaag 120

<210> 176

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 176

ccctcccggt ttaatgcttg cgtccaatca gaacttgcga gcagcttggg ggtgctcccc 60

accaccaccc cctttcaggg cagcacttct ttcatttgtt tgcctgaaat atcctgtttt 120

<210> 177

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 177

tctccccctt ctcggggcgc taaagagctc agagaaatct gcattttaat tgacattcca 60

gggaaagaga gaggggcgcc tgattgcttt tctattcaaa ctaaacaggc ttttcagatg 120

<210> 178

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 178

aaaatgagtc gagcccggtg gaagatttac agggatttgg gaccagggag acaatggagt 60

tttaaatgtt gatagcgttt cacaccaaac acataatggg tgccctgagt ttgaaagcag 120

<210> 179

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 179

gtttgccttt aagggtaggt tttattgaat tgggagaaag gaaggagggt ctggagaacg 60

gtctgcaagg gagcggggtg cgaggagagc gacggaggct ggtggtcccc gagcgggcct 120

<210> 180

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 180

cctgcgccag gtccgtggtc cgcgagggtc ggagccagcg ccacgcaggg ccctcggctt 60

ttttccccag cgcggccccg tggccgaaaa aaatgagcgc gcaagttaga actcgcctgc 120

<210> 181

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 181

gctttcgaca tctcccagcc gctgcggcag agcaggcagg cgaactagaa aaagtcggtc 60

cagttcgccc cctagcccca ggcgcactgc gctccctcca gcgccgacgg cggctgcggg 120

<210> 182

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 182

cgaggggcgc ccaggcaccg gagccgtcgc ctccccagca gtgccacccc tggccatcgc 60

cacccctacg ggatgtcacc ggccccaccc cagcggcccc ctcccgaaaa aggggtctcc 120

<210> 183

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 183

gcagggcctg gatggtacgg aagttcccag gacacacgga tggggcacct cggagatgac 60

cccgggaacc ggcctgcggg gctgtgccac gggcaggcga accttccgcc cgtctgctct 120

<210> 184

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 184

ggggtcggag aaacgcgccc agtcagcggg aatgcgcccg ccagggtttg acagggaggg 60

gggcgtcaac gccagaagca gatgtcagca tctggatggg ggtagcagag gcagaacttc 120

<210> 185

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 185

cagaacgccg gccgtcagca cgcacccacc tcccccatca cctatgcagc cacagaggga 60

ggacacggag cgtcctccac aatgctagtc cttccgcgcc ccccgcccgg aggacggtgc 120

<210> 186

<211> 120

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 186

acccggcgtc ccgggctccc gtggcgccca cgtccccggc tggcgaagct ccccgaggcg 60

tcttgcgcgg cgattacctt gaatggcaac gctcctccgc gacctgtgtt gtccccgcgc 120

Claims (7)

1. A DNA probe library for hybridization to FGFR1, FGFR2, or FGFR3 genes; the method is characterized in that: the DNA probe comprises sequences shown in SEQ ID NO.1 to SEQ ID NO. 186.

2. A method for enriching and detecting FGFR1, FGFR2 or FGFR3 gene fragments, which is characterized in that: the method comprises the following steps:

step 1: extracting genomic DNA from a human cell, plasma, tissue or FFPE sample, thereby obtaining fragmented double-stranded DNA as a DNA sample library;

step 2: designing a DNA probe which is hybridized with FGFR1, FGFR2 or FGFR3 gene as a DNA probe library aiming at the FGFR1, FGFR2 or FGFR3 gene to be enriched; the DNA probe library is the DNA probe library of claim 1;

and step 3: hybridizing the DNA sample library with the DNA probe library;

and 4, step 4: separating the hybridization product of step 3, and then releasing the gene fragments enriched by hybridization;

and 5: and (3) carrying out detection of fusion variation on the enriched FGFR1, FGFR2 or FGFR3 gene fragments.

3. The method for enriching and detecting the FGFR1, FGFR2 or FGFR3 gene fragments of claim 2, wherein: the DNA sample pool in the step 1 is composed of double-stranded DNA fragments, and the step 1 includes extracting whole genomic DNA of a subject and then fragmenting it; wherein the subject is human and the whole genomic DNA is extracted from a cell, plasma, tissue or FFPE sample of the subject.

4. The method for enriching and detecting the FGFR1, FGFR2 or FGFR3 gene fragments of claim 2, wherein: in the step 5, the fusion mutation detection of the enriched FGFR1, FGFR2 or FGFR3 gene fragments adopts next generation sequencing technology NGS.

5. The application of a DNA probe library hybridized with FGFR1, FGFR2 or FGFR3 genes in detecting related fusion variation of FGFR1, FGFR2 or FGFR3 genes; the method is characterized in that: the application comprises the method of claim 2.

6. A kit for enriching FGFR1, FGFR2 or FGFR3 gene fragments, characterized in that the kit comprises the DNA probe library of claim 1.

7. A kit for detecting fusion variation of FGFR1, FGFR2 or FGFR3 gene fragments, which comprises the DNA probe library of claim 1.

Images