CN107904234A - A kind of clupeoid chondrioid genom sequence universal primer and design and amplification method - Google Patents

Abstract

The present invention relates to a kind of clupeoid chondrioid genom sequence universal primer that is expanded, can efficiently and accurately be identified Clupoidea fish by mitochondrial genome complete sequence and it is related to and amplification method, universal primer for the amplification of clupeoid chondrioid genom sequence can be with the mitochondrial genome complete sequence of a variety of Clupoidea fish of efficient amplification, and is further used for the research that Clupoidea fish different classifications hierarchy evolves and classifies；The adjacent primer extension product of universal primer of the clupeoid chondrioid genom sequence amplification provided ensures the overlapping region for having 50 bp or so, it is higher to splice accuracy, the mitochondrial genome complete sequence of a variety of Clupoidea fish can be amplified with rapid batch, and some independent species design specific primer need not be directed to, the DNA profiling of high quality is not required, time cost is saved, and is greatly improved work efficiency, there is efficient quick.

Description

A kind of clupeoid chondrioid genom sequence universal primer and design and amplification Method

Technical field

The present invention relates to a kind of field of molecular biotechnology, more particularly to one kind to be expanded by mitochondrial genome complete sequence Increase, efficiently and accurately Clupoidea fish can be identified clupeoid chondrioid genom sequence universal primer and relate to And and amplification method.

Background technology

Clupoidea species is various, and many types form is closely similar, brings great puzzlement for Identification of Species, and it is being lost Also there is nearer affiliation in coming into, it is necessary to which especially measure mitochondrial genome complete sequence ability is sequenced on a large scale Carry out efficient system developmental analysis.

If by designing the universal primer of clupeoid chondrioid genom sequence, can disposably expand in batches The mitochondria full-length genome of a variety of Clupoidea fish, it is not necessary to design specific primer for some independent species, just saving Also greatly improved work efficiency while time cost, there is efficient quick.Reflect for the species of Clupoidea fish Fixed, phyletic evolution, plasm resource protection and Sustainable Development and Utilization provide important Molecular tools and data accumulation.

Patent Office of the People's Republic of China disclosed a kind of spider mitochondrial genome complete sequence amplimer and expansion on the 29th in September in 2017 The invention mandate of increasing method, authorizes publication number：CN104531688B, the invention are passed through using the STb gene of extraction spider genome Long PCR (L-PCR) amplifications and sequencing of mitochondrial genomes DNA identifies spider have with the present invention and play the same tune on different musical instruments it It is wonderful, but it is only capable of specific amplification spider mitochondrial DNA, it is impossible to multiple species are once expanded in batches, need to be set for particular species Specific amplimer is counted, and the Clupoidea fish universal primer of this patent design can expand clupeoid genoid in batches Group complete sequence, it is easy to operate, and time-saving and efficiency.

The content of the invention

Various to solve above-mentioned Clupoidea species, many types form is closely similar, is brought for Identification of Species greatly tired The problem of disturbing, is expanded, can be efficiently and accurately to Clupoidea fish the present invention provides one kind by mitochondrial genome complete sequence The clupeoid chondrioid genom sequence universal primer identified and it is related to and amplification method, being capable of disposable batch Expand the mitochondria full-length genome of a variety of Clupoidea fish, it is not necessary to design specific primer for some independent species, just exist Also greatly improved work efficiency while saving time cost, there is efficient quick.

To achieve the above object, the present invention uses following technical scheme：

A kind of clupeoid chondrioid genom sequence universal primer, the clupeoid chondrioid genome total order Row universal primer includes ten pair of primers：Wherein,

Pair of primers：

Forward primer 1 is as shown in SEQ ID NO.1：5’ - GCAYAAAGGTTTGGTCCT - 3’；

Reverse primer 1 is as shown in SEQ ID NO.2：5’ - TCGTTGAACAAACGAACCCTT - 3’；

Second pair of primer：

Forward primer 2 is as shown in SEQ ID NO.3：5’ - GTTTACGACCTCGATGTTGG - 3’；

Reverse primer 2 is as shown in SEQ ID NO.4：5’ - AGTGTCTGRGTTGCATYCAG - 3’；

3rd pair of primer：

Forward primer 3 is as shown in SEQ ID NO.5：5’ - AGACCATGAGCCTTCAAAG - 3’；

Reverse primer 3 is as shown in SEQ ID NO.6：5’ - TARCTTCARTATCATTGRTGKCC - 3’；

4th pair of primer：

Forward primer 4 is as shown in SEQ ID NO.7：5’ - CCACTCTGMCACTTTCTT - 3’；

Reverse primer 4 is as shown in SEQ ID NO.8：5’ - TGTGCTTGGTGGGCCATT - 3’；

5th pair of primer：

Forward primer 5 is as shown in SEQ ID NO.9：5’ - ATTCAACTAATYGCHACAGC - 3’；

Reverse primer 5 is as shown in SEQ ID NO.10：5’ - GANAGNGGGTCAATBGCYATRTA - 3’；

6th pair of primer：

Forward primer 6 is as shown in SEQ ID NO.11：5’ - CTACAATGCTAAAARTYCTHATCCC - 3’；

Reverse primer 6 is as shown in SEQ ID NO.12：5’ - TAGCCYHTGCTTGGATTTGCACCAAGAGT - 3’；

7th pair of primer：

Forward primer 7 is as shown in SEQ ID NO.13：5’ - GAAGGATAAYAGTCATCCGYTGGTCTTAGG - 3’；

Reverse primer 7 is as shown in SEQ ID NO.14：5’ - GADGTDGAGAATGCTACRAT - 3’；

8th pair of primer：

Forward primer 8 is as shown in SEQ ID NO.15：5’ - GTYACAGCYAACAAYATRTTYCARYT - 3’；

Reverse primer 8 is as shown in SEQ ID NO.16：5’ - CCAAAGTTTCATCADACTGA - 3’；

9th pair of primer：

Forward primer 9 is as shown in SEQ ID NO.17：5’ - CCRAGACTAATGACTTGAAA - 3’；

Reverse primer 9 is as shown in SEQ ID NO.18：5’ - ATAGTTTAANTTBAGAATNCTAGCTTTGGG - 3’；

Tenth pair of primer：

Forward primer 10 is as shown in SEQ ID NO.19：5’ - TCGAGCACCCCTTTATTATTATTGG - 3’；

Reverse primer 10 is as shown in SEQ ID NO.20：5’ - TAGTACTCCACCATTAACTTATGCG - 3’；

Tenth pair of primers：

Forward primer 11 is as shown in SEQ ID NO.21：5’ - CATATCATGAATGATAAGATCA - 3’；

Reverse primer 11 is as shown in SEQ ID NO.22：5’ - TCTCCATAACCACTCTTTACGC - 3’.

A kind of design method of clupeoid chondrioid genom sequence universal primer, from Genbank databases All clupeoid chondrioid genom sequences announced at present are downloaded, multiple alignment is carried out to these sequences, is looked for Go out the region more larger than more conservative region and variation, in ten a pair of of universal primer of conservative region design.

Preferably, there is the overlay region of 45~55bp between the adjacent primer extension product of described ten a pair of of universal primers Domain.

A kind of amplification method of clupeoid chondrioid genom sequence universal primer, the Clupoidea fish line grain The amplification method of body genom sequence universal primer comprises the following steps：

（1）DNA extraction agent boxes method extracts the STb gene of Clupoidea fish；

（2）Using the clupeoid chondrioid genom sequence universal primer group described in claim 1；

（3）Using Clupoidea fish total genomic dna as masterplate, PCR amplification is carried out；

（4）1.0% agarose gel electrophoresis of amplimer, RNA isolation kit are tapped and recovered amplified fragments, sequencing and sequence assembly.

Preferably, step（1）The Genome DNA extraction of the Clupoidea fish, to avoid intestinal contents from polluting, casts out Head belly, only extracts DNA from its afterbody.

Preferably, step（3）The reaction system of the PCR amplification forms：DNTP2 μ L, the 10 × TaqDNA of 2.5mM Polymerase Buffer2.5 μ L, respectively take each forward primer and each reverse primer, the TaqDNA polymerizations of 5U/ μ L that 1 μ L concentration is 10 μM 0.2 μ L of enzyme, 100g/ μ L 1 μ L of DNA masterplates solution and add to 25 μ L with sterilizing distilled water；

The PCR amplification condition is：95 DEG C of pre-degenerations 5min, subsequent 95 DEG C of denaturation 30s, 56 DEG C of annealing 30s, 72 DEG C of extensions 1~ 4min, carries out 35 circulations altogether, finally carries out 72 DEG C and extends 10min, is preserved under the conditions of 4 DEG C.

Preferably, the step（4）Middle gel electrophoresis uses 1.0% agarose gel electrophoresis, is tapped and recovered after sequencing respectively Splicing can obtain mitochondrial genome complete sequence to sequence from beginning to end successively.

By the universal primer for designing clupeoid chondrioid genom sequence, it is not necessary to for some independent species Specific primer is designed, amplified production is in 1000-4000 bp or so, it is not necessary to the DNA profiling of high quality；Can disposably it criticize Amount expands the mitochondria full-length genome of a variety of Clupoidea fish, not only saves time cost, but also greatly improves work efficiency, and has There is the characteristics of efficient quick.

The beneficial effects of the invention are as follows：

1）A kind of universal primer for the amplification of clupeoid chondrioid genom sequence is provided, can be more with efficient amplification The mitochondrial genome complete sequence of kind of Clupoidea fish, and be further used for Clupoidea fish different classifications hierarchy evolve and The research of classification；

2）The adjacent primer extension product of universal primer of the clupeoid chondrioid genom sequence amplification provided ensures There is the overlapping region of 50 bp or so, splicing accuracy is higher, and the mitochondria of a variety of Clupoidea fish can be amplified with rapid batch Genom sequence；

3）Some independent species design specific primer need not be directed to, it is not necessary to the DNA profiling of high quality；

4）The mitochondria full-length genome of a variety of Clupoidea fish can be disposably expanded in batches, not only save time cost, but also greatly Improve work efficiency greatly, there is efficient quick.

Brief description of the drawings

Fig. 1 is that the universal primer of clupeoid chondrioid genom sequence provided by the present invention amplification expands spot Ji Agarose gel electrophoretogram afterwards；

Fig. 2 is after the universal primer of clupeoid chondrioid genom sequence provided by the present invention amplification expands knife long-tailed anchovy Agarose gel electrophoretogram；

Fig. 3 is after the universal primer of clupeoid chondrioid genom sequence provided by the present invention amplification expands long-tail anchovy Agarose gel electrophoretogram；

Fig. 4 is that the universal primer of clupeoid chondrioid genom sequence provided by the present invention amplification expands Han Shi ribs Anchovy Agarose gel electrophoretogram afterwards；

Fig. 5 is after the universal primer of clupeoid chondrioid genom sequence provided by the present invention amplification expands black mouth Chinese herring Agarose gel electrophoretogram；

Fig. 6 is after the universal primer of clupeoid chondrioid genom sequence provided by the present invention amplification expands yellow crucian carp Agarose gel electrophoretogram；

Fig. 7 is that the universal primer of clupeoid chondrioid genom sequence provided by the present invention amplification expands the fine jade after Chinese herring Sepharose electrophoresis pattern；

Fig. 8 is that the universal primer of clupeoid chondrioid genom sequence provided by the present invention amplification expands Bangladesh hilsa herring Agarose gel electrophoretogram afterwards；

Fig. 9 is that the universal primer of clupeoid chondrioid genom sequence provided by the present invention amplification expands China little Sha Agarose gel electrophoretogram after fourth fish；

Wherein, M is the Marker of DL5000, and 1~11 swimming lane is followed successively by the amplification of primer pair 1~11, and N swimming lanes are DL2000 Marker.

Embodiment

The technical solution in the embodiment of the present invention is clearly and completely retouched with reference to the embodiment of the present invention and attached drawing State, it is clear that described embodiment only part of the embodiment of the present invention, rather than whole embodiments.Based on the implementation in the present invention Example, those of ordinary skill in the art's all other embodiments obtained without making creative work, belongs to Protection scope of the present invention.

Embodiment：

（1）Collection, identification and the preservation of Clupoidea fish sample

Clupoidea fish sample used is picked up from the natural environment of field in embodiment, is adopted sample and is taken back laboratory and carries out letter Breath registration, is identified with stereomicroscope, to determine species.The good Clupoidea fish sample of Identification of Species with 100% absolute alcohol Soak and be stored in spare in 4 DEG C of refrigerator.The Clupoidea fish species gathered include：Spot Ji, knife long-tailed anchovy, long-tail anchovy, Han Shi ribs Anchovy, black mouth Chinese herring, yellow crucian carp, Chinese herring, Bangladesh hilsa herring and Chinese Sardinella.

（2）The extraction of clupeoid chondrioid gene

The total genomic dna of Clupoidea fish sample to be measured is extracted using DNA extraction agent boxes method（Including karyogene and mitochondria Gene）, to avoid intestinal contents from polluting, cast out its incidence, only extract DNA from its afterbody.DNA extractions directly use DNA Extracts kit is extracted, and the total genomic dna of extraction is spare under the conditions of being stored in -20 DEG C.

（3）The design and synthesis of clupeoid chondrioid genome universal primer

All clupeoid chondrioid genom sequences announced at present are downloaded from Genbank databases, to this A little sequences carry out multiple alignment, find out the region more larger than more conservative region and variation, and it is a pair of to be related to ten in conservative region Universal primer.Synthesized universal primer is：

Pair of primers：

Forward primer 1 is as shown in SEQ ID NO.1：5’ - GCAYAAAGGTTTGGTCCT - 3’；

Reverse primer 1 is as shown in SEQ ID NO.2：5’ - TCGTTGAACAAACGAACCCTT - 3’；

Second pair of primer：

Forward primer 2 is as shown in SEQ ID NO.3：5’ - GTTTACGACCTCGATGTTGG - 3’；

Reverse primer 2 is as shown in SEQ ID NO.4：5’ - AGTGTCTGRGTTGCATYCAG - 3’；

3rd pair of primer：

Forward primer 3 is as shown in SEQ ID NO.5：5’ - AGACCATGAGCCTTCAAAG - 3’；

Reverse primer 3 is as shown in SEQ ID NO.6：5’ - TARCTTCARTATCATTGRTGKCC - 3’；

4th pair of primer：

Forward primer 4 is as shown in SEQ ID NO.7：5’ - CCACTCTGMCACTTTCTT - 3’；

Reverse primer 4 is as shown in SEQ ID NO.8：5’ - TGTGCTTGGTGGGCCATT - 3’；

5th pair of primer：

Forward primer 5 is as shown in SEQ ID NO.9：5’ - ATTCAACTAATYGCHACAGC - 3’；

Reverse primer 5 is as shown in SEQ ID NO.10：5’ - GANAGNGGGTCAATBGCYATRTA - 3’；

6th pair of primer：

Forward primer 6 is as shown in SEQ ID NO.11：5’ - CTACAATGCTAAAARTYCTHATCCC - 3’；

Reverse primer 6 is as shown in SEQ ID NO.12：5’ - TAGCCYHTGCTTGGATTTGCACCAAGAGT - 3’；

7th pair of primer：

Forward primer 7 is as shown in SEQ ID NO.13：5’ - GAAGGATAAYAGTCATCCGYTGGTCTTAGG - 3’；

Reverse primer 7 is as shown in SEQ ID NO.14：5’ - GADGTDGAGAATGCTACRAT - 3’；

8th pair of primer：

Forward primer 8 is as shown in SEQ ID NO.15：5’ - GTYACAGCYAACAAYATRTTYCARYT - 3’；

Reverse primer 8 is as shown in SEQ ID NO.16：5’ - CCAAAGTTTCATCADACTGA - 3’；

9th pair of primer：

Forward primer 9 is as shown in SEQ ID NO.17：5’ - CCRAGACTAATGACTTGAAA - 3’；

Reverse primer 9 is as shown in SEQ ID NO.18：5’ - ATAGTTTAANTTBAGAATNCTAGCTTTGGG - 3’；

Tenth pair of primer：

Forward primer 10 is as shown in SEQ ID NO.19：5’ - TCGAGCACCCCTTTATTATTATTGG - 3’；

Reverse primer 10 is as shown in SEQ ID NO.20：5’ - TAGTACTCCACCATTAACTTATGCG - 3’；

Tenth pair of primers：

Forward primer 11 is as shown in SEQ ID NO.21：5’ - CATATCATGAATGATAAGATCA - 3’；

Reverse primer 11 is as shown in SEQ ID NO.22：5’ - TCTCCATAACCACTCTTTACGC - 3’.

（4）PCR amplification mitochondrial genomes

The reaction system of PCR amplification forms：DNTP2 μ L of 2.5mM, 10 × Taq DNA polymerase Buffer2.5 μ L, respectively take 1 μ L concentration is 10 μM of each forward primer and each reverse primer, the 0.2 μ L of Taq DNA polymerase of 5U/ μ L, the DNA masterplates of 100g/ μ L 1 μ L of solution and add to 25 μ L with sterilizing distilled water；

The PCR amplification condition is：95 DEG C of pre-degenerations 5min, subsequent 95 DEG C of denaturation 30s, 56 DEG C of annealing 30s, 72 DEG C of extensions 1~ 4min, carries out 35 circulations altogether, finally carries out 72 DEG C and extends 10min, is preserved under the conditions of 4 DEG C.

（5）The sequencing of PCR product

1.0% agarose gel electrophoresis of PCR product, gel electrophoresis spectrum as shown in Fig. 1~9, obtain size 1000~ Fragment between 4000 bp, the fragment expanded are tapped and recovered using RNA isolation kit, to the Chinese little Sha fourths in recovery product Fish, spot Ji and long-tail anchovy are sequenced, and compared with sequence of the same race on GenBank, sequence similarity exceedes sequencing result 99.9%, it further demonstrate that the universal primer can actually effectively expand clupeoid chondrioid genome.Chinese little Sha fourths Fish（Sardinella nymphaea）Sequencing result as shown in sequence table SEQ ID NO.23, spot Ji (Konosirus Punctatus sequencing result) is as shown in sequence table SEQ ID NO.24, the sequencing result of long-tail anchovy (Coilia mystus) As shown in sequence table SEQ ID NO.25.

Sequence table

<110>Zhejiang Ocean university

<120>A kind of clupeoid chondrioid genom sequence universal primer and it is related to and amplification method

<160> 25

<170> SIPOSequenceListing 1.0

<210> 1

<211> 18

<212> DNA

<213>Universal primer (consensus primer)

<400> 1

gcayaaaggt ttggtcct 18

<210> 2

<211> 21

<212> DNA

<213>Universal primer (consensus primer)

<400> 2

tcgttgaaca aacgaaccct t 21

<210> 3

<211> 20

<212> DNA

<213>Universal primer (consensus primer)

<400> 3

gtttacgacc tcgatgttgg 20

<210> 4

<211> 20

<212> DNA

<213>Universal primer (consensus primer)

<400> 4

agtgtctgrg ttgcatycag 20

<210> 5

<211> 19

<212> DNA

<213>Universal primer (consensus primer)

<400> 5

agaccatgag ccttcaaag 19

<210> 6

<211> 23

<212> DNA

<213>Universal primer (consensus primer)

<400> 6

tarcttcart atcattgrtg kcc 23

<210> 7

<211> 18

<212> DNA

<213>Universal primer (consensus primer)

<400> 7

ccactctgmc actttctt 18

<210> 8

<211> 18

<212> DNA

<213>Universal primer (consensus primer)

<400> 8

tgtgcttggt gggccatt 18

<210> 9

<211> 20

<212> DNA

<213>Universal primer (consensus primer)

<400> 9

attcaactaa tygchacagc 20

<210> 10

<211> 23

<212> DNA

<213>Universal primer (consensus primer)

<400> 10

ganagngggt caatbgcyat rta 23

<210> 11

<211> 25

<212> DNA

<213>Universal primer (consensus primer)

<400> 11

ctacaatgct aaaartycth atccc 25

<210> 12

<211> 29

<212> DNA

<213>Universal primer (consensus primer)

<400> 12

tagccyhtgc ttggatttgc accaagagt 29

<210> 13

<211> 30

<212> DNA

<213>Universal primer (consensus primer)

<400> 13

gaaggataay agtcatccgy tggtcttagg 30

<210> 14

<211> 20

<212> DNA

<213>Universal primer (consensus primer)

<400> 14

gadgtdgaga atgctacrat 20

<210> 15

<211> 26

<212> DNA

<213>Universal primer (consensus primer)

<400> 15

gtyacagcya acaayatrtt ycaryt 26

<210> 16

<211> 20

<212> DNA

<213>Universal primer (consensus primer)

<400> 16

ccaaagtttc atcadactga 20

<210> 17

<211> 20

<212> DNA

<213>Universal primer (consensus primer)

<400> 17

ccragactaa tgacttgaaa 20

<210> 18

<211> 30

<212> DNA

<213>Universal primer (consensus primer)

<400> 18

atagtttaan ttbagaatnc tagctttggg 30

<210> 19

<211> 25

<212> DNA

<213>Universal primer (consensus primer)

<400> 19

tcgagcaccc ctttattatt attgg 25

<210> 20

<211> 25

<212> DNA

<213>Universal primer (consensus primer)

<400> 20

tagtactcca ccattaactt atgcg 25

<210> 21

<211> 22

<212> DNA

<213>Universal primer (consensus primer)

<400> 21

catatcatga atgataagat ca 22

<210> 22

<211> 22

<212> DNA

<213>Universal primer (consensus primer)

<400> 22

tctccataac cactctttac gc 22

<210> 23

<211> 16616

<212> DNA

<213>Sequencing result (the The sequencing results of Sardinella of Chinese Sardinella nymphaea)

<400> 23

gctggtgtag cttaaacaaa gcataacact gaagatgtta agatgggccg tgataagccc 60

cacaagcact gaaaggtttg gtcctggctt tattatcagc tttaccccaa tttacacatg 120

cgagcctccg cggccccgtg aggatgccct cagcctcccg tccggagatg aggagcgggc 180

atcaggcacg atgttcggcc catgacgcct tgtttagcca cacccccaag ggaattcagc 240

agtgattgac attaagacat aagtgaaagc ttgacttagt tagggatgag cagagccggt 300

aaatctcgtg ccagccaccg cggttatacg agaggctcga gttgataatc tcggcgtaaa 360

gagtggttat ggagaagact aaactaaagc tgaagacccc ccaggctgtt taacgcatgc 420

gggtgttcga accacttata cgaaagtagc tttaacgcat tccaccagaa tccacgacag 480

ctgggaaaca aactgggatt agatacccca ctatgccctg ccgtaaactt agatattaca 540

accacaacga atatccgcct gaggactacg agcactagct taaaactcaa aggacttggc 600

ggtgcttcag acccccctag aggagcctgt tctataaccg ataacccccg ttcaacctca 660

ctacttcttg ctcattccgt ttatataccg ccgtcgccag cttatcctgt gaaggttcaa 720

cattaagcaa aatgggtaag gcccagaacg tcaggtcgag gtgcagcgta cgaagtagga 780

agaaatgggc tacattatct gaaacagatt attcacggag agtcgcctga aaccgagcgc 840

tctaaaggtg gatttagcag taagggggga gtagagcgcc cccctgaagc cggctctgaa 900

gcgcgcacac accgcccgtc gttctccccg attaagcaac atattgtaac taacacaaca 960

acaaccagca aggggaggta agtcgtaaca tggtaagtgt accggaaggt gcacttggaa 1020

taaccaggat gtggctgaga tagtcaagca tctcccttac accgagaaaa catccatgca 1080

aattggatca tcctgagcca atcagctagc tcataacccc caacaactaa aataacaata 1140

tacataccat aacacaaact taataactaa aactaaatca ttcaaccacc ccagtatggg 1200

cgacaaaaaa ggatttcacc gaagccatag ataaagtacc gcaagggaaa gctgaaaaag 1260

aactgaaaaa atgcactcaa gcacaaaaaa gcagagacta aaactcgtac cttttgcatc 1320

atgatctagc cagtaccccc aagcaaagag aactccagtt tgatgccccg aaacctccga 1380

cgagctaccc cagggccgcc taaaacaggg caaacccatc tctgtggcaa aagagtggaa 1440

agaccctcag gtagaggtga aaagcctacc gagtcaggtt atagctggtt gcccagaaaa 1500

tgaataatag ttcagcccca tacaacgccc cccgccaacc ataatagtcc cacaggcaac 1560

agggccaccc atgggagtta gttaaaggag gtacagctcc cttaacaaag gatacaacct 1620

tactcaggag gctaaggaat acatcaaatg aaagcacagg tttcagtggg cctaaaagca 1680

gccatctgta cagaaagcgt caaagctcat accagctcaa gcttattata atgttaagca 1740

ccttcctatg cccctcaaca atactgagcc ctcctatgct cccataggag agaccctgct 1800

agaacgagta ataagaagca agaacttctc cctgcacatg tgtaagtcga gctggaccca 1860

ccgtcgacaa ctaacgaacc caacggaaga ggacaatgcg ttaccgccac tataggccaa 1920

ggagaacacg cacctcaccc catcgttaac cccacacagg agtgccttag tcacgggaaa 1980

gacttaaaga ataaaaagga actcggcaaa cccagacccc gcctgtttac caaaaacatc 2040

gcctcctgcc cctaattata taggaggtcc cgcctgccct gtgaccaaaa gtttaacggc 2100

cgcggtatca taaccgtgca aaggtagcgc aatcaattgt cttttaaatg gggacctgta 2160

tgaatggcat aacgagggtc tagctgtctc ttttttctgg tcaatgaaac tgatctgccc 2220

gtgcagaagc gggcatgcta acacaagacg agaagaccct atggagcttt agacgcccac 2280

caaccgcgag cagcgctccc cttaacgggg gcccaaacaa cgcggccctg gtataaacgt 2340

cttaggttgg ggcgaccgca ggggagaaca aagcccctga gagggttggg aacggcccta 2400

aaactgagag ctacagctcc aagtcgcaaa acgtttgacc gataatgatc cggcttactg 2460

ccgatcaacg gaccaagtta ccctagggat aacagcgcaa tcctctccca gagcccatat 2520

cgacgagggg gtttacgacc tcgatgttgg atcaggacat cctaatggtg cagccgctat 2580

taagggttcg tttgttcaac gattaaagtc ctacgtgatc tgagttcaga ccggagtaat 2640

ccaggtcagt ttctatctgt gacgccacct ttcctagtac gaaaggaccg gaatgatgag 2700

gcccatacct ttggcacgcc tccctcccaa cccgctgaag acaactaaag cgggataggg 2760

gagcaactta ccgcccgaga aaagggcatg tttaggtggc agagcccggt aagtgcagga 2820

agcctaagcc ttccctttca gaggttcaag tcctctccca aactatgctt cacaccattc 2880

taacgcacat cattaacccc ctagcctata tcgtccctgt tcttctggct gtggctttcc 2940

ttacgctgat tgaacggaag gttctgggct atatgcaatt gcgaaaggga cctaacgtgg 3000

taggccctta cgggctcctc cagcccattg cagacggagt gaaactcttt atcaaagagc 3060

cggtccgccc ctctacgtcc tccccctttt tatttttagc aacccccact ctcgcgctga 3120

cgctagcgct cactctatga gcccctcttc ccctgcccca cccagttacg gacctaaacc 3180

tgggcctgct ttttatccta gcaatctcca gcttagccgt ctactcaatt ttaggctccg 3240

ggtgagcatc aaactccaaa tacgccctaa ttggagccct gcgggcggtg gctcaaacca 3300

tctcctacga ggtagcactt gggcttattc tcctttgcac ggtcgtcttt acaggaggct 3360

ttaccctctc catgtttagt accacccaag aagcgatctg actcttagcc cctgcttgac 3420

ccctcgcagc aatgtggtac gtctctacct tagcggaaac caaccgagca ccctttgacc 3480

taaccgaggg agaatcagaa ctagtatccg ggtttaacgt ggagtacgcg gggggccctt 3540

tcgccctatt cttcctcgca gagtacgcca acatcttatt tataaacacc ctgtccagca 3600

ttctgtttat tggagcctcc catttcccat ccctacccga gctcaccacg atcaatatta 3660

taatcaaggc cgccctcctt tcagcagtgt tcctctgagt ccgggcttcg taccctcgat 3720

tccgatatga ccagcttata cacctggtgt gaaagagctt cctccccctc accctggcgc 3780

ttatcctctg acatatctcg cttccggtcg gaaccgcggg gcttcctcct caactctaac 3840

cctataggga gctgtgcctg aacgcctaag ggccactttg atagagtgaa ccaagggggt 3900

tagactcccc ccggctcctt agaaagaagg ggctcgaacc cgtcctgcag agatcaaaac 3960

tctgggtgct tccactacac cactttctag caaggtcagc taaataagct ctcgggccca 4020

taccccgaac acgttggtta gaatccttcc cctgctaatg aacccccacg tattaaccgt 4080

cttgctgatt agccttggac tgggaaccac tctgactttc gccagctccc attgacttct 4140

ggcgtgaatg ggcctggaaa tcaacaccct cgccattatt ccccttatag cgcgccgcca 4200

gcacccccga gcggttgagg ctacgaccaa gtacttcttg actcaagcca ccgctgccgc 4260

catgatcttg tttgctgcca caactaacgc gtgagtctgc gggcaatggg aaatcaccca 4320

cttgacaagc cccaccgcct gtacaattgc cgttacagct ctagcactca agattggcct 4380

tgctccagcc cacttttgaa tgcctgaagt actccaggga attaccctgc ccaccggact 4440

aattctctcc acctgacaaa aactagcacc cttcgcccta atcgtccagg tagccggcaa 4500

cgctcacccc tacttactaa cagcactagc catctgctct acactggtcg ggggctgagg 4560

gggactcaac caaactcagc tccggaaggt actagcctac tcatcaattg cccacctagg 4620

ttggatgatt ctcgtggccc aaatggcccc tcacatgacc gtgctggccc tagccaccta 4680

tatcatcata acaacagcgg ccttcctcac cctatacaaa atgaactcga ccaaaaccgt 4740

cactctcgcc tcagccttta ccaaggcccc cacccttgcg gccctcgcct gcttaatttt 4800

actttccctc gggggtctcc cccctcttac cgggtttata cctaagtgac ttatcctgca 4860

ggaggtttcc aaccaaggat tttcccttac agcaaccgtt attgccctga cagctctcct 4920

cagcttgtac ttctaccttc gtctgaccta tgcggtagca cttaccttct ccccctacac 4980

cacctgttct accacaccat gacgagcccc gactagccac ccaacccttc tgttatctgc 5040

atccgtcgtg ctagcgacgt gcttgttacc cctgaccccc tccatcctcg ggattgtagc 5100

ttagggactt aggaataact aagaccaaga gccttcaaag ctctaagtag gagtgagact 5160

ctcctagtcc ctgatctaag acctgcaaga ctctatctta catctcctga atgcaactca 5220

gacactttaa ttaagccaag gcctctctag acgggaaggc ctcgatccta caagatctta 5280

gttaacagct aagcgcccaa accagcgagc atccgcctat cttaccccgc ctgccggggc 5340

gagggcgggg aaagccccgg caggctgtta acctgcgtct tcaggtttgc aacctgacat 5400

gaacttcacc acagagcttt tttgataaga aaaggaatta aacctttatc ctcggagcta 5460

caatccgccg cctaaacctt cggccatcct atctgtggca atcacacgtt gatttttttc 5520

aactaatcat aaagatattg gcacccttta tctagtattc ggtgcctgag cagggatggt 5580

cggaaccgcc ctaagtcttc taatccgagc ggagctgagc cagccagggg cactccttgg 5640

agatgaccag atttataacg tcattgtcac cgcacatgct ttcgtaatga ttttctttat 5700

agttatgcca atcctgattg gagggtttgg aaactgactt gttcctctaa tgatcggagc 5760

gcccgacatg gccttcccgc gaatgaacaa catgagcttc tggctccttc ctccttcttt 5820

ccttcttctc ctcgcctctt caggcgtaga agccggagca gggacaggct gaacagtgta 5880

cccgccctta gcaggtaatc tagcccacgc cggtgcctct gttgacctaa ccattttctc 5940

actacacctg gcaggtattt catcaattct aggggcgatt aacttcatca ccacaatcat 6000

taacatgaaa cctcctgcaa tctcgcagta ccagacaccc ctgtttgtct gagctgtcct 6060

tgtaacagct gttcttctac ttctctctct cccagtccta gctgctggaa ttaccatgct 6120

cctgaccgac cgaaacctga acacgacttt cttcgatcct gcaggcggag gggacccaat 6180

cctgtaccaa cacctatttt gattcttcgg gcatccggag gtatatattc ttattcttcc 6240

tgggttcggg atgatctccc acatcgtagc ttactatgct gggaaaaaag agcccttcgg 6300

atatatggga atggtttgag caatgatggc tatcggcctc ctaggcttca tcgtttgggc 6360

ccaccacatg ttcacagttg gaatggacgt tgacacacgg gcttacttca cgtcggcaac 6420

catgatcatc gctatcccga ccggggtaaa agtgtttagc tggcttgcca cccttcacgg 6480

aggctcaatc aaatgagaga cccctctcct ttgagctctt ggctttatct tcttattcac 6540

agttgggggt ctaacaggaa ttgtactatc caattcttcc ttagacatta tcctccacga 6600

cacctactac gttgtagcac acttccacta tgtcttatcg atgggtgcag tgttcgctat 6660

tatggccgcc tttgtgcact gattccctct cttctcaggg tacacgctac acagcacttg 6720

aacgaaagcc cacttcgcgg ttatgttcgt aggggtcaat cttaccttct tcccacagca 6780

cttcctgggc ctagcaggca tgccacgacg gtactccgac tacccagacg cctataccct 6840

ttgaaacacc gtatcctcaa ttggatcgct gatctccctt gtggccgtga ttatgttctt 6900

gtttattctt tgagaggcat tcaccgctaa acgagaagtt gcatcagtag agctgactac 6960

gacaaacgta gagtggctgc acggctgccc tcctccatac cacaccttcg aggaaccagc 7020

ctttgtacaa gtgcaggcaa agtaacgaga aaggagggaa ttgaaccccc gtcagatggt 7080

ttcaagccaa ctgcatggcc actctgccac tttcttgatt tagacgctag taaaactatt 7140

accttgcctt gtcaaggcaa aattgtgggt taaacccccg cgtgtcttgc tcagagctaa 7200

gtggcacatc cctcacaact aggattgcaa gacgcggcct ccccggttat agaagaactc 7260

ctgcacttcc acgaccacgc tctaatgatt gtccttctaa ttagtgtttt ggtcctttac 7320

atcattgtat ctatagtttc aaccaaactt acagacaagt acatcctaga ctcacaagaa 7380

attgagattg tgtgaaccat cctgccggct gttgttctga tcatgatcgc gttaccttct 7440

ttacggattc tctatctagt agatgaaatt aacgaccccc acctaaccat taaagccatg 7500

ggccaccagt ggtactgaag ctatgagtac accgactacg aagacttggg ctttgactcc 7560

tacatggtcc caactcagga tcttattcca ggccaattcc gactcttgga gacggaccat 7620

cgaatagtcc ttccgataga gtcccccatc cgaatcctcg tctcagcaga agacgtattg 7680

cactcgtgag ccgttcccgc actcggagta aaaatggacg cggttccagg acgacttaac 7740

caaactgcct ttattgcctc ccgcccaggg gtcttctacg gacaatgctc ggaaatttgt 7800

ggtgcaaatc acagtttcat gccaattgta gtagaagctg tacctctaaa acacttcgaa 7860

gactgatcct cactaatact tgaagacgcc tcactaggaa gctaaactgg gcctagcgtt 7920

agccttttaa gctgaagatt ggtgaccccc aaccacccct agtgacatgc ctcaattgaa 7980

tccggcccct tgatttgcaa ttctcgtctt ctcctgattg atcttcctaa cagtaattcc 8040

gccaaaagtt atggcccata attttaataa cgaaccaacc actgcggggg ccgaaaaggc 8100

taaacccgaa ccttgaacct gaccatgata ctaagctttt ttgaccaatt tataagcccc 8160

acctacatgg gaatccccct catcgcacta gcaattgcac tgccctgagc actataccca 8220

acccccacct cccgctgatt aaacaatcga gtgctcaccc ttcagggatg gtttattaac 8280

cgttttaccc agcaacttct tctgccttta aatgttggtg gccataagtg agcacttctg 8340

cttacgtctc taatggtttt cctatttacc attaacatgc taggccttct gccctatacg 8400

ttcacaccaa caacccagct atcgcttaat atgggacttg ctgttcccct ttgactcgcc 8460

acagtgatca tcggtatgcg taataagcca accgatgccc taggccacct cctcccagaa 8520

ggtacaccaa tcgcccttat tccagtacta attattatcg aaaccatcag cttatttatt 8580

cgaccgctgg cactgggggt ccgattgacc gccaacctca ctgctggtca cttacttatc 8640

cagctaattg gcacagcagc atttactctt atgacttcga tgcctattgt tgccgtttta 8700

actgccactg tcctcttcct actcaccctg ctagaagttg ctgtagcgat gatccaggca 8760

tacgtctttg tattgcttct aagcctctat cttcaagaga acgtctaatg gcccaccaag 8820

cgcacgcatt tcacatagta gaccccagcc cctggcccct taccggagca gtcggtgcac 8880

tgctgctaac ctccggcacc gcaatttgat tccacttcca atcaactgcc gtaataacac 8940

tagggttagt tctaaccctt ctaactatat accagtgatg acgagacgtc gtacgagaag 9000

gcactttcca agggcaccac acccctccag tccaaaaggg ccttcgatac gggataatcc 9060

tgtttattac atcagaggtt ttcttctttg caggcttttt ttgagccttc taccactcta 9120

gcctcgcacc tactcccgaa ctgggaggtt gctggccccc aaccggcgta actaccctcg 9180

acccgttcga agtcccccta cttaacacag cggtcctttt ggcctcaggt gttaccgtca 9240

catgagctca ccacagcatt atggaggggg agcgagccca ggcgatccag tctctcaccc 9300

tgacaattct tctcggcttc tattttacat tcctgcaagc cctggagtac tacgaggcac 9360

ctttcaccat tgcagatggc gtatacggct ctactttctt cgtagccaca ggattccacg 9420

gccttcacgt aatcatcggg tcaacattct tagctgtctg ccttctacgc caagtattct 9480

accatttcac ctcgagccac cactttgggt tcgaggcagc tgcatggtac tgacactttg 9540

ttgacgtggt ttgactcttc ctttatgtct ctatctactg atgaggttca taatctttct 9600

agtataaaga atagtacagg tggcttccaa ccatctaatc ttggttaaat tccaaggaaa 9660

gataatgagt ttaattataa cagtcatgac aatcgcatcc cttctatcaa ttattctagt 9720

actcgtctcg ttctggctac cccagatagc gcctgactcg gagaaactct ccccctacga 9780

atgtggtttt gacccactgg ggtctgctcg gctaccgttt tccctacgtt tcttcctagt 9840

ggcaatcctg ttcctactat ttgacctaga aatcgccctt ctacttcccc taccctgagg 9900

taatcaatta cttaacccgc tacttactgt ctcctgggcc accgcaatcc tggttctact 9960

tacacttgga ctagtttatg agtgaacgca gggtggacta gagtgagccg aatagggaat 10020

tagtccaact taagactcct gatttcggct cagatgatta tggttaaatt ccataattcc 10080

cttatgacac cagtacattt tagctttagc ggagcattcg ttcttggcct ggtgggatta 10140

gcattccacc gcgctcacct tctttccgcc ctcctctgcc tggaaggaat gatactgtcc 10200

ttatttatgg ggttctcact atggacactc cagacggggg ttgccaactt ctctgcagcc 10260

ccaatacttc tcctcgcctt ctctgcctgt gaagccagtg cgggcttagc cctgctcgta 10320

gctactgccc gaactcacgg gacagatcgg ctgcagagcc ttaacctcct ccaatgttaa 10380

aagtcctcat cccaaccctc atactgcttc caacaacctg atttgcgcct aaaaagtgac 10440

tttgaactgc tgcaatcaca cacagcctca ccattgcctt actaagcctt acctggctaa 10500

actggacaac ggaaacgggc tgagccctca ccaacaccta tatagccatt gatcccttat 10560

cctccccgct gttggttctt acatgctggc tccttcctct aatggtttta gccagtcaaa 10620

atcacactcg aacagagcca atctcgcggc aacgcatgtt tattagcctg cttgcctccc 10680

tgcaggcgtt ccttattcta gcatttgggg ctacagaaat tatcatgttc tacgtcatat 10740

ttgaggcaac tctggtacca acgctaatca ttatcacccg ctgaggaaat caagcagagc 10800

gtttgaatgc cggcacctac ttcttgtttt ataccttagc gggctcgctt ccgctactag 10860

tcgccctatt aaccctgcaa agctcaacgg gcaccctttc aataatcaca ctaaattttg 10920

gccaagcact ctcgttagtc tcctgagccg acaaggcctg atgagcgggc tgcctgctgg 10980

ccttcttggt taagatacca ctctacggcg ttcacttgtg gctgcccaag gcccacgtag 11040

aagccccaat tgccgggtcc atagttctgg cagcggttct actaaaacta gggggttacg 11100

gcatgattcg aataacctcc attttagacc ccctaaccaa ggacatggcg taccccttca 11160

ttgtgcttgc cctctgaggg gtaattataa ctgggtccat ctgtttacgc caaactgacc 11220

ttaaatcgct cattgcttat tcatctgtta gccacatggg tctagtggcc ggaggtatct 11280

taattcaaac accctgggga ctaaccgggg ccatcatttt aatgattgct cacggactag 11340

tatcctcagc cctcttttgt ctggcaaaca cggcctacga gcggacccac agccgtacaa 11400

tagtcctagc acggggatta cagatgctct tccccttgac agccacttga tgattcgttg 11460

ctaacctggc caaccttgca ctccctcctc tccctaatct tatgggagag gtaataatta 11520

ttaccacgat gttcaactgg tcgccttgaa cgctggtact gacgggagtc ggaaccctaa 11580

ttacagccgg gtactcgctg tacatattcc tgataaccca acgagggccc gtaccagccc 11640

acatcgtagg acttaccccc tatcacaccc gagaacacct cctaatctcg cttcatttaa 11700

tccccgtcat cctgcttatg ttcaagccag aattcgtgtg gggatgattc tactgcagat 11760

atggtttaac aaaaatgttg gattgtgatt ccaaagacgg gagttcaaac ctccttatct 11820

gccgagagag gcttggagca atagagactg ctaatctcta cctccggagt taaaatctcc 11880

ggctccctcg gcctttgaag gataatagtt catccgctgg tcttaggaac cagacactct 11940

tggtgcaaat ctcaagcaaa ggctatgcag acgaccttga ttttaacgtc ctcacttacc 12000

cttatttttg gtcttttaac ctacccaatc ttaacaacga ttaaccccac gcccaaaagc 12060

cctgactggg ccgtgacaca cgtcaaaacc gcagttagct ccgcattttt agttagcctt 12120

ctgccattgt ttatttttct cgaccaaggc gtggagacaa tcgtcaccac atgacactga 12180

ataaacacgg gcacctttaa cattaacatt agcctaaaat ttgatgccta ctcaatcatt 12240

tttaccccta ttgctctcta tgttacatga tcgattttag aatttgcctc atgatacatg 12300

cacgcagatc cttatgtgag tcgtttcttc aagtacctcc ttatgttttt gattgccatg 12360

attatcctag taacagccaa caacatgttc cagcttttta ttggttggga aggtgtgggt 12420

atcatatcgt tcctcttgat cgggtgatgg tacggccgag ctgacgccaa caccgctgcc 12480

ctacaagccg tcctctataa ccgagttggg gatattggac taattatagc catggcatgg 12540

ttcgctataa acctaaactc gtgagagatg caacaaattt tttcgctctc ccacaccagc 12600

acaaccctcc cccttctagg cctaattgtg gccgctaccg ggaagtcggc ccagttcgga 12660

ctccacccct gactaccctc tgcaatggag ggccccactc cagtttccgc tcttctgcac 12720

tccagcacta tggtcgtggc tgggatcttc ctgcttatcc ggcttcaccc acttacccac 12780

tccaatcaaa ccgcactaac tatctgtcta tgccttggag ctcttactac cttatttaca 12840

gccacctgcg ctttaaccca gaatgacatc aagaaaatcg tagcattttc cacatctagt 12900

cagctgggcc ttatgatggt gaccatcggc cttgaccaac cccaactagc ctttttccac 12960

atctgcaccc acgccttctt caaggcaatg ctcttccttt gctccggatc tatcatccac 13020

agccttaatg atgagcaaga catccgtaaa atagggggaa tacacaatct ggccccattc 13080

acctccacct gcctgactat tggcagcctt gccctcacag ggaccccctt ccttgctggc 13140

ttcttctcaa aagatgcgat catcgaagcc ctaaacacct catacctaaa cgcctgagcc 13200

ctagcactca ccttaatcgc cacctccttt acagcggtct atagcttccg agtcgtcttc 13260

ttcgtaacca tggggtcacc tcggttcacc gcattcgccc ccattaacga gaatgacccg 13320

gcggtaatca atcctatcaa acgtctcgcc tgagggagca ttgtcgcagg tcttattctt 13380

acctcaaatg ttcttcccac aaagacccca atcatgacca tacccccact gctgaaacta 13440

gccgccctcg cggttacaat tgtgggcctc ctggtggcca tagaactagc tactcttacc 13500

tccaaacaat tcaaaccaac cccaattatt aagctacaca acttctcgaa tatactaggg 13560

tacttcccgg ccgtagtaca ccgcctggcc cccaagctta acctagtttt aggccaaaca 13620

atggccaacc aaatggtcga tcaggcctga tttgaggctg cagggccaaa ggggctagcc 13680

tccactcaac agaagatatc cgccactacg agtgacgccc agcgaggact aatcaagacg 13740

tacttgataa tcttcttgat aacttcaggt ctggctactc tgcttgcctc tgcctaggga 13800

gctcgtagag ccccccgagc ccgacctcgg gacacctcaa atactactat tagggttaag 13860

aaaagcaccc atgcgcagaa gaggattaat gccccaccaa cggagtatat caccacaact 13920

cccatgtcca cccgctcaat agggaattcc tccacccctg cggcgtagtc gtaaactcac 13980

ggacgcatca tcactaccat caagatcgtc cccacaataa ggactattgt tcgcccaaac 14040

acctggttat ccccaagagt ctcagggtga ggatccgccg acaaagcagc tgagaacccg 14100

aagaccacta acattccccc cagataaact aagaacaaga ctaaagccaa gaacggcgcc 14160

ccctgagcag ataaaatcgc gcaacacgtc cctgaacaca gcaccaggcc aagtgccgcg 14220

aaataagggg ccggattagc ggccaccaac accatcccta acaaaaaccc gtagagtgtg 14280

caaatgataa ttaattccac agtttctacc cgcattttaa ccgagactaa tgacttgaaa 14340

aaccaccgtt gttgttcaac catagaaact ctttaatggc aagcctgcgg aaatcacacc 14400

ctttgctgaa gattgctaac gacgcagttg ttgaccttcc agccccctcc aacatttcgg 14460

tctgatgaaa tttcggatca ctcctgggcc tatgcttagc atcccagatc ctaacaggac 14520

tattcttagc catgcactac acctcagata tcgcaaccgc cttctcttct gttgcacaca 14580

tctgtcgtga cgtaaactac ggttgactaa ttcgtaatat gcatgcaaac ggagcatctt 14640

tctttttcat ctgcatttat gctcacatcg gtcgaggcct ttactacggc tcctacttat 14700

ataaggagac ctgaaacatc ggagtagtcc tcctgctcct tactatgatg acagcattcg 14760

tagggtatgt tttaccttga ggacagatgt ctttctgagg tgctacagtc attactaacc 14820

tcctttcagc cgttccttac gtcggggaga tgctagttca atgaatttga ggcggcttct 14880

cggttgacaa tgcaacttta actcgattct ttgccttcca cttcctcttc ccattcgtga 14940

ttgcaggtgc tactgtcctt catcttctct tcctgcatga aacaggctcc aacaacccgg 15000

caggccttaa ctcagacgct gacaaaatct cattccaccc atacttctca tacaaagacc 15060

tgctaggctt cgccgttata cttcttgcgc taacttctct tgccctattc tccccaaacc 15120

ttctggggga cccggacaac ttcacccccg ccaaccctct agtcacacct ccccatatta 15180

aaccggagtg atacttcctc tttgcctacg ctatcctacg gtcgatccct aacaagcttg 15240

gaggggtctt agccctccta ttctcaattc ttgtccttat ggtcgtccca atcctgcaca 15300

cctcgaagca gcgaggcctt accttccgac caatgactca attcttgttc tgaacattaa 15360

tcgcagatgt ggctattctg acgtgaattg ggggcatgcc cgttgagcac ccatttgtta 15420

tcgtcggcca agtagcctca gtgctttact ttgcattatt cctagttctt gccccactag 15480

cagggtgagc agagaataaa gctttagagt gaaactgcct cagtagctta gccttaaagc 15540

gccggtcttg taaaccggag atcggaggtt aaaatccccc ctaaggccca gagaagaaag 15600

attctaactt ccacctctga ctcccaaagc tagaattcta agttaaacta ttctctgagt 15660

cgcacttcca catccacgcc ccaatgttag aattctagat taaattatgc tctggtttta 15720

ataagatcgc tgtccctagt taagccatca gaccatcata cagtgtttaa ggttagccct 15780

gacgaacgct tacgccgtgc ctctactata tacattgcac agtacgacct gattgtaagc 15840

gcaacgagag agctgcctta acggccctaa ctggagatta gaccctagcc ggactttccc 15900

acccacctgt ctcacggccc tacgggggcc ggaatgcacc agcgtacctg aacagataac 15960

atcaaaagca ggggccaact cattaaacag agccaccctg gctgaccact gagcaccccc 16020

atagaatgaa caagcaggcg ggttattcgg accgaaggcc gcgcgaatcg tccagggcga 16080

ccacaacgtc gatctcgacc gggcggcagt taggaacact gccccagatc accactgcct 16140

ggttagttgc agtaccccct tgacgcccca gctggtaatg tttatttaaa gtgccaccag 16200

acgtggcaag ccgcgtgggc ggtctgggat ctatatagga caccctttcg ttataaagac 16260

ttagcggagg aaccctgttt aacagggcgg gatagaaggt ctggtatata acgctcagcc 16320

gctagcgcat gccgggcatt catttagatg catttggctc ttttatttta gggtcacttt 16380

catcttgcat ttggcgactc cctcctaatg ctaacgtcaa aggttgtaca ttttcttgct 16440

tgttatacta attatgccta cactcaagga cataaatgta agaaccacac aagtgacgtc 16500

aggtgcatag agcatcatta cttatcaaca tttcctgctt ttactacgct gattagacta 16560

gaaagctttt atagtcgaca aacccccccc tacccccccc gccaatgaga atgtta 16616

<210> 24

<211> 16704

<212> DNA

<213>The sequencing result (The sequencing results of Konosirus punctatus) of spot Ji

<400> 24

gcttgtgtag cttaaaacaa agcacgacac tgaagatgtc gagatggacc ttgaaaagtc 60

ccgtgagcac aaaggtttgg tcctgacttt attatcagct ttaactcaat ttacacatgc 120

aagcctccgc gaccctgtga ggatgccctc aatctcccgt ccggagacga ggagccggta 180

tcaggcacaa tgttttgccc aagacgcctt gcttagccac acccctaagg gaattcagca 240

gtgataaaca ttaagccata agtgaaaact tgacttagtc agagttaaga gggccggtaa 300

aactcgtgcc agccaccgcg gttatacgag agacccaagt tgataagccc ggcgtaaaga 360

gtggttatgg atagcacaaa actaaagcta aagaccccct aggctgttat acgcacctgg 420

cggctcgagt caccaacacg aaagtagctt taccccctcc gaccagaacc cacgacagct 480

ggggcacaaa ctgggattag ataccccact atgccctgcc gtaaacttag atattccact 540

acaacaaata tccgcctggg gactacgagc gctagcttaa aacccaaagg acttggcggt 600

gcttcagaac cccctagagg agcctgttct agaaccgata acccccgttc aacctcacta 660

ctccttgctt ttcccgccta tataccgccg tcgccagctt accctatgaa ggtaccacag 720

taagcagagt gggcaatgcc caaaacgtca ggtcgaggtg tagcgtacgg agtaggaaga 780

aatgggctac attatctgaa acagataatt cacggaagga tgcctgaaat cggcaacccg 840

aaggtggatt tagcagtaaa gggggaatag agcgccccct tgaagccggc tctgaagcgc 900

gcacacaccg cccgtcactc tccccgacaa cacccacact tgatacctaa cgcaacaatc 960

agccgaaagg ggaggcaagt cgtaacatgg taagtgtacc ggaaggtgca cttggaataa 1020

ccagggtgtg gctgagatag ttaagcgcct cccttacacc gagaagacat ccatgcaagt 1080

tggatcgccc tgaaccaaac agctagctca aacaaaaaga ctaaatttac aacataggct 1140

aatctaacat aaactaatta cgccaaccaa accatttgac ccccccagta cgggcgacgg 1200

aaaaggacta atgaagcgat agataaagta ccgcaaggga aagctgaaag agagctgaaa 1260

tagcgcactg aagtgaataa aagcagagac cagacctcgt accttttgca tcatgatcta 1320

gccagtaaac tcaagcgaag agccctctag tttgaacccc cgaaaccgga cgagctaccc 1380

cgggacagcc tattgtaggg ccaacccgtc tctgtggcaa aagagtggga agatccccgg 1440

gtagaggtga aagacctacc gagtcaggtt atagctggtt gcccaggaaa cgaatagaag 1500

ttcagccccg tatcagccct tatccacagc agttttactg aagttaggca ggaggacaac 1560

cgcgggagtt agttgaagga ggtacagctc cctcaacaaa ggacacaacc tttacaggag 1620

gctaagagat atactaaatt aaggccacag gtttcagtgg gcctaaaagc agccacctga 1680

acagaaagcg ttaaagctca gaccagatta agcctattat accattaaat atcctcaatg 1740

cccctactaa tactgggccc ccctatgctc acataggaga gaccatgcta gaacgagtaa 1800

taagaagaat gaacttctcc cagcacatgt gtaagtcgaa tcggaccccc catcgacaat 1860

taacgaaccc aaaaacagag ggccatgcat ccgtgccata ctaggccaag aaaatcacgc 1920

agaccccatc gttgacccca cacaggagtg ctcacacaag gaaagactta aaaaatgaaa 1980

aggaactcgg caaacctgaa ccccgcctgt ttaccaaaaa catcgcctcc tgctcacatc 2040

tgttatagga ggtcccgcct gccctgtgac caaaagttta acggccgcgg tattttaacc 2100

gtgcaaaggt agcgcaatca attgtctttt aaatggagac ctgtatgaat ggcataacga 2160

gggtttaact gtctcttttt ttcggtcaat gaaactgatc tgcccgtgca gaagcgggca 2220

tacccacaca agacgagaag accctatgga gctttagacg ctcaccaacc actaaaagag 2280

acccgatcta acgggctttc aaacaatgtg gccgtggtac aaacgtcttc ggttggggcg 2340

accgcggagg agagaaaagc ctccgagagg attgggggag accctaaaac caagagccac 2400

agctctaagt cacaaaatat ttgaccgaca acgatccggc ttcttgccga tcaacggacc 2460

aagttaccct agggataaca gcgcaatccc ctcccagagt ccatatcgac gagggggttt 2520

acgacctcga tgttggatca ggacatccta atggtgcagc cgctattaag ggttcgtttg 2580

ttcaacgatt aaagtcctac gtgatctgag ttcagaccgg agtaatccag gtcagtttct 2640

atctgtgatg ccacccttcc tagtacgaaa ggaccggaat gatgaggcct atgctccaag 2700

cacgcctccc cccgacctga tgaaaacaac tgaagcaggc aaaggggggc caatccccgg 2760

cccgagagaa gggcgagttg gggtggcaga gcccggtaaa tgcaggaagc ctaagccttc 2820

cccttcaggg gttcaaatcc tctccccagc tatgctccac attattatag cccacatcat 2880

taaccctctt gcctatattg ttcccgtgct tctagcggtg gccttcttaa cattaattga 2940

gcgaaaggtg ctcggctata tgcaattgcg caagggccct aacgtggtag ggccctacgg 3000

gcttctacaa cctattgcag acggggtaaa actatttatt aaagagccaa tccgcccgtc 3060

gacatcgtcc ccccttctat ttttagccgc cccggcttta gcgctaaccc tggccctcac 3120

cctatgggca cccttaccgc tcccctatcc agttacagat cttagccttg gtatgttgtt 3180

tattctggcc ctatctagcc tagccgtcta ctccatccta ggttcaggtt gagcatccaa 3240

ctccaagtat gcactgatcg gcgccctacg agccgtagcc caaaccatct cctacgaggt 3300

cgccctagga ctaatccttc tatgcacaat tgtactcacc ggagggttta ccctatcaat 3360

gtttagcacg acccaggaag gcatctgact tctggcccct gcctgaccct tggcagcaat 3420

gtgatacatt tccaccctag cagagactaa ccgtgccccg tttgacctca ctgaaggaga 3480

gtcggagttg gtctccgggt ttaatgtaga atatgcagga ggccccttcg ccctattttt 3540

ccttgccgaa tatgccaaca ttttattcat gaatacactg tccgccatcc tgttcatagg 3600

tgcctcgcat tttccctccc tgccagaatt aaccacaatc agcattatga ttaaagccgc 3660

cctcctctca ggagtattcc tctgggtccg agcctcatat ccgcgattcc gatatgatca 3720

gttgatgcac ctagtgtgga agaatttcct cccactcaca ctcgcacttg ttctatgaca 3780

tatctcgctg ccagtcggca ctgcggggct tccaccacaa ctctagtaaa cctaggagct 3840

gtgcctgaac gcttaagggc cactttgata gagtgaacca agggggttag actccccccg 3900

gctccttaga aagaagggat tcgaacccgt cctccagaga tcaaaactct gggtgcttcc 3960

gctacaccac ttcctagtaa ggtcagctaa ataagctttc gggcccatac cccgaacatg 4020

ttggttaaag tccttcccct gctaatgaac ccttatgtac ttactattct cctatttagc 4080

ctcgggctag gcaccacact gactttcgcc agctcccact gactcctagc ctgaatgggc 4140

ctagagatta acaccttagc catcatcccg ctcatagccc accagcatca cccgcgggcc 4200

gtcgaagcga ctactaagta cttcctcacg caagccgcag ctgccgctgt aatcctcttt 4260

gccagcacca cgaatgcatg gattactgga caatgagaga ttacccacct ctcgcaccca 4320

gtcgcctctg ccattgcggt gacagccctg gccctaaaga ttggacttgc ccccacacac 4380

ctttggctgc cagaagtgct ccaggggatt acactcacca ccggtcttat tctttccacc 4440

tggcaaaaac tggcccccct tgccctcatt ttacaagtag cagaaaatgc ccacccatac 4500

ctgttgacca ctctggcgat tacctccact cttgttggcg ggtgaggagg ccttaaccaa 4560

acccaactac ggaagatcct tgcatactcc tcagtcgcac acttaggatg gataatcctg 4620

gtagcccaga tggcccctca aatgactctg ttggcactga ttacatacat tgttatgacg 4680

accgcagcct ttctgacctt ggacaagata gaatctacca agaccattac tttggcatcg 4740

gcttggacca aggcccctac tttgagtgca ctggcctgcc tcattcttct ttctcttggg 4800

ggactcccgc ctcttacggg cttcatgcct aagtgattta ttctccaaga ggtttctagc 4860

caagggtttc ccctcacagc cacagtaatc gcactcacag ccctcttgag cctatatttt 4920

tacctccgcc tcacctacgc aataaccctc accctcgccc cttacaccat caattcttcc 4980

accccctggc gaagtcccgc taagcgaccc acactactcc tatccgcagc agttgtatca 5040

gcgacctgcc tcttacccct caccccctcc gctctagctc tgctggccta ggggcttagg 5100

atagcacata gaccgtgagc cttcaaagct ccaagcagga gtgaaaatct cctagcccct 5160

gataagactt gcaaggtttt atctcacatc ttctgaatgc aacccagaca ctttaattaa 5220

gctaaagcct ttctagatgg gaaggcctcg atcctacaaa atcttagtta acagctaagc 5280

gccctaacca gcgagcatcc atctactttc ccccgccgtc cggggactaa gggcgggggg 5340

aaagccccgg caggcgttaa cctgcgtctt caggtttgca acctgacatg aacttcacca 5400

cagagcttct tggtaagaag aggagttaaa cctctgtact cggagctaca atccgccgcc 5460

taaaccttcg gccatcctac ctgtggcaat tacacgttga tttttctcaa ctaatcataa 5520

agatattggc accctttatc tagtatttgg tgcctgagca ggaatagtag ggactgccct 5580

aagcctccta atccgagcgg aacttagcca gcccggcgcg ctcctaggag acgatcaaat 5640

ctacaatgtt atcgttacgg cacacgcctt tgtaatgatt ttcttcatag taatgccaat 5700

cctgattgga gggtttggga actgattggt tcccctaatg atcggggcac ccgacatggc 5760

attcccgcga atgaataaca tgagcttctg acttcttcct ccctctttcc ttctcctctt 5820

ggcctcctcc ggtgtagaag ctggggcggg gacaggatgg acagtctacc cccctttatc 5880

agggaaccta gcccatgcag gtgcatccgt cgacctaacc atcttctctc tccatcttgc 5940

aggtatttca tcgatcctag gggcaatcaa ttttattacc acaattatta atatgaaacc 6000

ccctgcaatc tcgcaatacc aaactccttt attcgtttgg gccgtgcttg tcactgctgt 6060

attacttctg ctatctcttc cggtgctggc tgcgggaatc actatgcttc taacggaccg 6120

gaatcttaat accaccttct tcgatcctgc tggcggagga gacccaatcc tttatcaaca 6180

cctcttctgg ttcttcgggc accctgaagt ctacattctt atcctgcctg gatttggaat 6240

gatttcccac atcgtagcct actactcagg caaaaaagag cccttcggct atatgggtat 6300

ggtgtgagca atgatggcca ttggactttt aggcttcatc gtgtgagccc accacatgtt 6360

cacagtaggc atggacgttg acactcgagc ttatttcacc tcagcaacaa tgattattgc 6420

catcccaaca ggggtaaaag tattcagctg acttgccaca ctgcatggag gctcaatcaa 6480

gtgagaaacg ccccttctct gggcccttgg attcatcttc ctcttcaccg tagggggtct 6540

aacagggatt gtgctgtcta attcctcgct agacattgtt ctccacgaca cctattacgt 6600

agtagcacac ttccactatg tcctgtcaat gggtgctgta tttgccatta tggctgcatt 6660

cgtgcactgg ttcccgctat tctcaggata taccctccac agcacttgaa caaaaatcca 6720

ttttggggtg atgtttgtgg gtgttaatct taccttcttc ccgcaacact tcctaggcct 6780

ggcaggaatg ccacggcgat actccgacta cccggacgcc tataccctgt gaaacacagt 6840

gtcttcgatc gggtcattaa tctctcttgt ggctgtaatt atgttcttat tcatcctgtg 6900

ggaagcattc gcagccaaac gagaagtttc gtcggtagaa ctaactataa caaatgtaga 6960

gtgactgcat ggctgccctc ctccatacca cacctttgaa gagccagctt ttgtacaagt 7020

acaagcaaaa taacgagaaa gggaggaatc gaacccccgt gagatggttt caagccagct 7080

gcatggccac tctgccactt tcttaaataa gacactagta aaaccattac cttgccttgt 7140

cgaggcaaaa ttgtgggtta aacccccgcg tgccttagct cagagcttaa tggcacatcc 7200

ctcacaacta ggattgcaag acgcggcctc ccctgttata gaagaactct tacacttcca 7260

cgaccacgcc ctaatgatcg ttttactgat tagcgtactg gtcctttaca ttattgtctc 7320

aatggtgtca accaagctca ccgataaata tatcctagat tcgcaagaaa tcgaaatcgt 7380

atgaactatc ctaccagccg tcatcctaat tatgatcgca ctcccttctc tgcggattct 7440

ttatctcatg gacgagatta atgacccaca cttaacaatc aaagccatgg gccaccaatg 7500

atattgaagc tacgagtata cagactacga agacctgggc ttcgactcgt acatggtgcc 7560

tactcaggat ctaacccctg gtcaattccg gctgctggaa acagaccatc gaatggtaat 7620

tcctatagaa tcgccgatcc gggtcctcgt ctcggctgaa gacgtacttc actcctgagc 7680

ggtccctgcc ctaggagtta aaatagacgc cgtaccaggc cgactaaatc aaactgcctt 7740

catcgcctct cgccccggag tcttctacgg gcaatgctcc gagatctgtg gtgccaatca 7800

cagttttatg cccatcgtag tagaagccgt cccactagaa cactttgaaa actgatcctc 7860

actaatactt gaagacgcct cactaggaag ctaaatcggg cctagcgcca gccttttaag 7920

ctggagactg gtggccccca accacctcta gtgacatgcc ccaacttaat cctgcccctt 7980

gatttgcaat tcttgtcttc tcctgactaa tcttcctaac ggtaattccc ccaaaagtcc 8040

tcgcccataa cttcaacaac gagcccacca ctgtaggggc cgagaaagcc aaacctgagc 8100

cctgaaactg accatgatac taagcttttt tgaccaattt ataagcccaa cacacctagg 8160

agtgccactc attgccctag cgattgcact cccttggacc ctttacccca cccccacctc 8220

ccgatggctc aacaaccggg tgctaactct ccaaggatga tttattaacc gttttactca 8280

gcaactcttg cttcccctaa accaaggcgg gcacaaatga gcagttctgc ttacttcgct 8340

tatggtcttc ctttttacca ttaatatgct tggactcctg ccctacacat ttacaccaac 8400

aactcaactt tctctaaaca tgggcttggc tgtgcccttg tgactcgcca cagtcatcat 8460

tggcatgcgc aaccagccca cagcagcttt aggacaccta ctgcccgagg gaacccccgt 8520

cccgcttatt ccagtcctta ttattatcga aacaattagc ctattcatcc gaccattggc 8580

actgggggtg cggttaactg ctaacctgac ggcgggccac ctgcttattc aactaatcgc 8640

cacggcagca tttgtactta tgcctatgat accaactgtt gctatcttga cagccactgt 8700

cttgtttctt ctcaccctac tagaagtagc tgtggccatg attcaagcat acgtatttgt 8760

tcttcttcta agtctatatc tacaagaaaa cgtctaatgg cccaccaagc acacgcattc 8820

cacatggtag acccaagccc ctgaccgcta accggagcag tcggcgctct gctgctgacc 8880

tctggcaccg caatttggtt ccacttccat tcaatcactt tgataaccct agggctagtg 8940

ctaactatcc taaccatata tcaatggtga cgagacgtcg ttcgagaggg aactttccaa 9000

ggacatcaca cacccccagt acaaaaaggg ctccgatacg gaatgatcct ctttattacc 9060

tcggaagtct tcttctttgc aggctttttc tgagctttct atcactctag cctagcccca 9120

actcctgaac taggagggtg ctggccccca acaggcatca ctactctcga cccctttgag 9180

gtccccttac tgaatacagc cgtcttacta gcctcaggcg ttaccgtcac atgagcccac 9240

cacagcctaa tggaaggaga acgtaaacaa gccatccaat ccctaaccct aactattctc 9300

ctaggcttct acttcacctt cttacaggcc ctagagtatt acgaagcccc cttcactatc 9360

gccgacggag tctacggctc tactttcttc gtagccacag ggttccacgg cctacacgtc 9420

attattggat ccacattcct ggccgtctgc cttatgcgcc aggtacttta ccactttacc 9480

tctaaccacc acttcgggtt tgaggcagct gcctgatact gacattttgt ggacgtggtc 9540

tgactcttcc tttacgtctc tatctattga tgaggatcat aacctttcta gtacaaaaga 9600

cagtataggt ggcttccaac cacttgatct tggttaaagc ccaaggaaag gtaatgagct 9660

taatcataac aatcctaaca atcacctcgc ttctatctat tattctggta atcgtatctt 9720

tctggctgcc ccagataaac ccggacgctg agaagctctc tccttatgag tgcgggtttg 9780

atcccctggg ctccgcccga ctcccctttt ccctacggtt ttttctcgtg gcaatccttt 9840

tcttgctctt tgacttagaa attgctctcc tcctgccgct tccctgaggt aaccagctgc 9900

tcaacccgct cactacagtg tcctgagcga ctgccatcct tgttctactc acgctcgggc 9960

tagtctacga gtgaacgcaa ggagggcttg agtgggctga ataggggatt agtccaacta 10020

aagacttctg atttcggctc agacaattat ggttaaagtc cataaccccc ttatgacacc 10080

agtacacttc agtttcagcg cagcatttat tcttggcctt ataggattag catttcatcg 10140

cacccacctt ctctcagcac ttttatgttt agagggaata atgctatccc tgtttatagc 10200

tctagccctg tggacactcc aaacagaggc caccaacctt actgcagcgc caatgcttct 10260

cctcgcgttc tccgcctgcg aagccagcac gggactggcc ctcctcgtgg ccacggctcg 10320

tactcatggc acagatcgac tccaaagcct taacctccta caatgctaaa gattctaatc 10380

cccactttaa tgctattccc aacaatctga ttaaccccta agaagtggct atggactgcc 10440

gcaatcaccc acagccttct catcgctttg cttagtctca cctgactcaa ctgaaccgcg 10500

gagacgggat ggaccacccc taacgcttac atggcaatcg accctctctc cgctccctta 10560

ttggtgctca cctgctgact gcttccctta atgattttgg caagccaaaa tcacacccga 10620

accgaacctc tctctcggca acgaatgtac atcagcctac tcgcttctct gcagactttc 10680

ctaatcctcg catttggagc cacagaaatt attatgtttt acattatatt tgaggcgaca 10740

ttggtcccaa cgttaattct gattactcgc tggggcaacc aggcagagcg actaaacgca 10800

ggcacctact tcttgttcta tactctcgcc ggatccctcc cgctactagt cgccttgcta 10860

acccttcaaa gctcgacggg aagtctttcc ataatcacgt taaacttcgg ccagccgcta 10920

acacttctat cctgagggga caaaatctgg tgggcaggat gccttattgc cttcctagtt 10980

aaaataccac tttacggggt ccacttatga ctacctaaag ctcacgtaga ggccccaatt 11040

gccgggtcaa tggtgctagc tgcagtcctg ttgaaactgg gaggctatgg catgatccga 11100

ataacatcca ttttggaccc gctcaccaag gaaatagcgt acccgtttat tgtgctcgcc 11160

ttgtggggca ttatcataac aggatcaatc tgcttgcgtc agacggactt gaagtcgcta 11220

atcgcttact cttcagtcag tcacatagga ctggtggccg gagggatttt gattcaaacc 11280

ccctgaggcc taaccggagc tattatttta ataattgccc acggactagt gtcatcagcc 11340

ctcttctgct tggcgaacac agcctatgaa cgcacgcaca gccgtaccat aattttagcc 11400

cgaggactac aaatactatt tcccctaacg gctacctgat ggtttattgc taatctagcc 11460

aaccttgcac tccccccact cccgaatctg atgggggaag ttatgattat cacaaccatg 11520

ttcaactgat ccccgtggac tcttatcctc acaggggttg gcacactaat tacggcaggc 11580

tactccctat acatgttcct catgacccaa cggggccccg tgccggcaca tattattgga 11640

ctaaccccct accacacacg agaacatctc ctcatcaccc ttcaccttgt acctgttatc 11700

cttctcattc tcaagccaga gttcatgtgg gggtggttct actgcaggta tagttttaac 11760

aaaaatgttg gattgtgatt ccaaggacag gagttcaaac ctccttaccc gccgagagag 11820

gcccgtagca gcagggactg ctaatctctg cccccgcagt taaactctgc ggctcactcg 11880

gcctctgaag gataatagtc atccgttggt cttaggaacc aaaaactctt ggtgcaaatc 11940

caagcagagg ctatgcaaac aaccctaatt ctgacatccg cacttaccct gatcttcgca 12000

ctgctggcct accccattat tactacaatc gaccccgcgg ctaaagacgc taactgggcc 12060

gtcacacatg tgaagactgc agtcagcacc gcatttgtgg ttagcctgct ccccttattt 12120

atcttcctgg atcaaggggt agagactatt gttaccactt gacactgaat aaatacatct 12180

actttcaaca tcaacatcag cctgaaattc gacgcctact ccatcatttt tacccctatt 12240

gccctctacg tcacgtggtc tatcctagaa ttcgcttcgt gatacataca ctcggacccg 12300

ttcataaacc gcttctttaa atacctacta atgtttctta ttgccatgat tatcctcgtc 12360

acagctaaca acatattcca actatttatt ggctgggagg gcgtgggaat tatatccttc 12420

ttactaattg gctggtgata tggtcgggct gatgctaata ctgccgcgct ccaagccgtg 12480

atctacaatc gagttggaga tattggacta attatgagca tagcgtgatt tgctataaat 12540

ttaaactcat gggagatgca gcagatcttc tccctctccc acaacactga tatgaccctg 12600

ccccttctgg ggctgattat tgctgcgacc ggcaaatcgg cgcaattcgg actccacccg 12660

tggctgcctt ccgcaatgga agggcccaca ccagtgtctg ccctactgca ttcaagcaca 12720

atggttgtag ccgggatctt cctcctcatt cgcctgcacc cccttactca ttctaatcaa 12780

acagccctta ccatctgttt gtgcctcgga gcactaacca cattattcac cgcaacctgc 12840

gccctcaccc agaatgacat taagaaaatc gtagcatttt ctacctctag ccagctaggg 12900

ctaatgatgg tcactattgg gctggatcaa cctcaactag ctttcttcca tatctgcacg 12960

cacgcttttt tcaaagctat attattcctc tgctctgggt ccatcatcca cagcctgaac 13020

gacgaacagg atatccgaaa aatagggggg atgcataacc tagccccctt cacttcaact 13080

tgtctcacca tcggtagcct ggccttaacc gggaccccct tccttgcggg gtttttctca 13140

aaagatgcaa tcattgaagc tttaaacact tcttatttaa acgcctgggc ccttgctctc 13200

accttgatcg cgacttcctt cactgcagtc tatagcttcc gggtcgtatt tttcgtcacc 13260

ataggaacgc ctcgattctc acctctttcc ccaattaatg aaaatgaccc cgcggtcatt 13320

aaccccatca aacgcctagc ctgaggcagc atcgtggcgg gtctaatcct tacctcaaat 13380

gctctcccta caaaaacccc tatcataacc atgccccctc ttcttaagct ggcggctctc 13440

gctgtcacta ttattggcct tctaacggct atggagctcg cctctctgac agccaaacag 13500

ttcaagccga ctccaattat caagctacac aacttctcta acatactagg ctacttcccc 13560

gcagtagtac accggttggc ccctaaacta aacctggtgc taggtcaaac gatggccaac 13620

cagttggttg atcagacatg attcgaggca gctgggccga aaggattagc ctcagcgcag 13680

ctcaagatat cggccgcgac tagcgatgcc caacggggaa tcatcaaaac atacctcatg 13740

atcttcctga ttacctcagg actcgccacc cttctggctt ccacctaagg agcccgcatt 13800

gatccgcggg atcgcccacg aacgagctct aggactacaa acagagttaa aagcagaact 13860

caggcgcaga acaatatcag catcccaccg accccataaa gaactgcgac tcctgcaaga 13920

tccccccgaa ccactaaaaa ctccttcacg gacgtcagca tccctacact aaactcataa 13980

gccggccgcc caaagtacaa tactgccacc accagcccta gaagataaaa catcgcatac 14040

tcgaacaccg ttacatccgt tcagtcttcg ggaaactcgt cagctgaaag agccgccgcg 14100

tacccaaaaa caacaagcat cccacccaga taaatcaaga atagagacaa ggcgaggaac 14160

ggtgtgccaa acattgctaa aacagcacaa cctgccgctg aagatagtac taaacccatg 14220

gccccaaagt aaggggacgg atatgaagca accccgacca aacccatcat gaacccagca 14280

agcaagagca ggatagtaga taccataaat ttctacccgg attttaaccg agactaatga 14340

cttgaaaaac caccgttgtt attcaactat agaaaccctt taatggcaag cctacgaaaa 14400

acccaccccc tcctaaaaat cgccaacgac gcagtagtcg acctcccggc cccctccaac 14460

atttcagttt gatgaaactt tggctcccta ctaggactat gtttagcagc acaaattctg 14520

accggactgt tcctagccat gcactacacc tctgacattg cgactgcctt ctcatctgtc 14580

acccatattt gccgcgacgt aaattacggc tgacttatcc gcaacatgca cgcaaacgga 14640

gcatccttct tctttatttg catctatgca cacattggcc gaggacttta ctacggctca 14700

tacctataca aggaaacctg aaatgtcgga gttgtcctgc tccttctagt catgataact 14760

gccttcgtcg gctacgtgct gccctgagga caaatgtcct tctgaggtgc tacagtgatt 14820

accaatctcc tatctgctgt accctacgtg ggagaagtac tagtccaatg gatctgggga 14880

ggcttctcgg tagacaatgc caccctcact cgattcttcg ccttccattt cttgttccca 14940

tttgtgattg caggagccac catcctgcac ctcctgttct tgcacgaaac agggtcaaac 15000

aaccccgcag gtcttaactc cgatgccgac aagatctcgt tccacccata cttttcctat 15060

aaagatttgc tgggcttcgc ggttatgctg ctagccttaa catctcttgc tctcttttct 15120

cctaatcttt taggcgaccc tgacaatttc acgcctgcca acccactggt gaccccacca 15180

catatcaagc cagaatggta cttcctgttt gcttacgcaa ttttacgatc tatccctaac 15240

aagctgggag gggtattagc tctgctattc tctatcctgg ttcttatggt tgtacccgtc 15300

ctgcacacct ctaagcaacg aggacttacg ttccgacctt taacacaatt cctgttctga 15360

acactagtgg cggatgtaat catccttacg tgaatcgggg gcatgcctgt tgaacacccc 15420

ttcattatca tcggccaagt ggcctcgcta atttattttt cattgttcct cgttcttgcc 15480

ccactagcgg gatgagtgga gaacaaggcc ttagaatgaa actgccccag tagcttagcc 15540

ttaaagcgcc ggtcttgtaa accggagacc ggaggttaaa atcctccctg aggcccagaa 15600

aagaaagatt ctaacttcca cctctaactc ccaaagctag aattctaaat taaactattc 15660

tctgacataa cccccttcct taccgtggtt tgagcgcgga gccacaagtg gagcttcacg 15720

cgtctacatt gatgccggat tcaatccgcg gcccagccta cggccccagg gcgcctccac 15780

catcggcctg acgcgcccaa aatgcactaa gtcacgcatg atggtgttaa tacattctat 15840

gtattatagt acatatataa tggtgtagta catactatgt attatcacca ataaattata 15900

ttaaccaaac aagtaagttg taaatcaaga cgtaccaaac cataacaatt aacatccaaa 15960

taaaattaaa atcaactaat ttgtactaag tccggtaaac aacgtgaatg gaacatgttc 16020

aatataatga atgaaaccta gtaaatcttc aacaaggcgc tgcattttcc atgcgttact 16080

caataaaaat cggtacgact agaattaatg tagtaagaac cgaccaacca accgagtaat 16140

tgcatatcat gaatgataag atcacggaca acaatcgtgg gggtttcacc cggtgaacta 16200

ttcctggcat ttggttccta tttcagggcc atacgtccat taatccccaa tgaatgaact 16260

ttccaggcat aagttaatgt tagagtacta tgtactcgtt acccaccatg ccgagcgttc 16320

acttatatgc atttggtatt tttttttccg ctcactttca ctttgcattt ggcgactcct 16380

tcctaatgtt aactttcagg gttgaacatt tccttgccta agttaacccc ttgtaaagct 16440

tcacagacat aacacaagaa ttgcataagt gatatcaagt gcataaactc ttaatactca 16500

ccccacagat ccttatacta tgcccccctt cttcgacaaa cgaccagagg ttttcgcgcg 16560

acaaacccct taccccctta cgccctaaga gccttattaa ttcatgtcaa accccaaaac 16620

catgaaagtc tcgactatgg cgtcttcagc gagttccgat acgtgctggt ctacatagtg 16680

ttacaaagtt aattctattg tgtc 16704

<210> 25

<211> 17142

<212> DNA

<213>The sequencing result (The sequencing results of Coilia mystus) of long-tail anchovy

<400> 25

gctcatgtag cttaaattaa agcatagcac tgaagatgct aagatgaacc ctaaaaagtt 60

ccacaagcat aaaggtttgg tcctagcctt agtatcagct ttaaccccac ttacacatgc 120

aagtctccgc aaccctgtga gaatgccccc aacttcccat acggaaatag ggagccggca 180

tcaggcacaa taacttagcc caaaacgcct tgctcagcca cacccccaag ggaattcagc 240

agtgacagac attaagccat aagtgaaaac ttgacttagt tagggttaag agggtcggtt 300

aaactcgtgc cagccaccgc ggttatacga gagaccctag ttgactcaca cggcgtaaag 360

agtggttatg gaattacaaa actaaagccg aaagcccccc agactgtcat acgcatccgg 420

gggccagaac tccactatac gaaagtagct ttaccagcgc ctaccagaac ccacgatagc 480

tggggcacaa actgggatta gataccccac tatgcccagc cataaacctt gatactctca 540

tacaattagt atccgccagg gaactacgag caccagctta aaacccaaag gacttggcgg 600

tgcctcagac ccccctagag gagcctgttc tagaaccgat aacccccgtt aaacctcacc 660

acttcttgcc tattccgcct atataccacc gtcgccagct taccctgtga agggaaaaaa 720

gtaagcaaaa tggaaacttt ccaaaacgtc aggtcgaggt gtagcatacg aagtgggaag 780

aaatgggcta cattacctga tacaggatat tcacggaaag tcacctgaaa cggtaattca 840

aaggtggatt tagcagtaaa agaggaatag agtgcccctt tgaagccggc tctgaggcgc 900

gcacacaccg cccgtcactc tccccaaaca cttacacacc aaagtaaata acatcataaa 960

acataacaag gggaggcaag tcgtaacatg gtaagtgtac cggaaggtgc acttggaata 1020

actaggacgt ggccgagata gttaggcaac tcccttacac tgagtccaca tccatgcaaa 1080

ttgggtcgtc ctaagctaga aagctagctc aaacatctta gcaaaacaaa acattttttt 1140

aactactcat atacaaaaca cccacaaacc aaatcatttg actaccccag tacgggcgac 1200

aaaaaaggat ccacgaagcc atagacaaag taccgcaagg gaaagctgaa aaagaaatga 1260

aaaaattcat taaagcacaa aaaagcagag acactcctcg taccttttgc atcatgattc 1320

agctagtaaa atacatgcaa agtgacctct agtttgtaac cccgaaaccc gacgagctac 1380

tccgggacag cctatcaagg gccaacccgt ctctgtggca aaagagtggg aagatccccg 1440

agtagaagca aaagacctac cgagccgggt tatagctggt tgcccaaata aatgaataaa 1500

agttcagccc cgtatagccc aactcacagc agttttacta caaaagacaa agagctatta 1560

agcgggagtt aatcagggga ggtacagctc ccctgataaa ggacacaacc ttataaagga 1620

gattaaggaa cataatcagt aaaggccaca ggtctcagtg ggcctgaaag cagccacccg 1680

accagaaagc gttaaagctc agaccaaaat aaccctatta ttatattaaa cactccaaaa 1740

tccctaacat aaatgggcct ttctatgccc ccatagaaga gattatgctg aaatgagtaa 1800

caagaagttt gaacttctcc ccgcacaagt gtaaatcgaa ttagacccac taccgacaat 1860

taacgaaccc aataacagag ggccccacat taccgccgta gaaggccagg aaaaacatgt 1920

ccccctaatc gttaacccca cacaggagtg ctaaaccaag gaaagactaa aagaaaaaaa 1980

aggaactcgg caaacctaaa ccccgcctgt ttaccaaaaa catcgcctct tgccaactct 2040

attgtataag aggtcccacc tgccctgtga ccaaaagttt aacggccgcg gtatcctaac 2100

cgtgcgaagg tagcgcaatc aattgccttt taaatgaagg cctgtatgaa tggtataacg 2160

agggtttaac tgtctctttt ttccagtcag ttaaattgat ctgtccgtgc agaagcggac 2220

ataattatac aagacgagaa gaccctatgg agctttagac actaaccaac catgaaaagc 2280

aataaattac atatacccca aacacccatg ggcctggcat aatagtctta ggttggggcg 2340

accacgggag ataacaaagc tcccgagcag atcagggaaa tcctgaaact aagagttaca 2400

actctaagtc acaaaaattt tgactgaaat gatccggcca aagccgatta acgaaccaag 2460

ttaccctagg gataacagcg caatcccctt tcagagtcca tatcgacaag ggggtttacg 2520

acctcgatgt tggatcagga catcctaatg gtgcagccgc tattaagggt tcgtttgttc 2580

aacgattaaa gtcctacgtg atctgagttc agaccggagt aatccaggtc ggtttctatc 2640

tgtaaagcaa ccacccctag tacgaaagga ccggggtggg gaggcccatg ttaataacaa 2700

gcctcccatc tacctgttga aaacagctaa aacagacaaa gatgggcacc ccccagaccc 2760

gcgaccaggg tcacgaaata attcgcgcta gggtggcaga gcccggtaaa tgcagaaagc 2820

ctaagccttt cataccggag gttcaaatcc tccccctaac tatgctaagc accatcatca 2880

cccacattat taacccttta gcttacattg tgcccgtact attagcagtt gccttcttaa 2940

cccttgtcga acgaaaagtt ctaggctata tacaactacg aaaaggccct aacgtagttg 3000

gtccttacgg actacttcag ccaattgcag acggagttaa actatttatc aaagagccca 3060

tccgaccgtc cacctcctcc ccctttttat ttttagctac ccccactctc gcacttaccc 3120

tagcactcac cttatgagct cccctcccca tgccctaccc tgttacagat ttaaacttaa 3180

gcatactatt tattctagca ctatcaagct tggccgtata ctctatttta ggctcaggat 3240

gagcatctaa ctccaaatat gcactaattg gtgctcttcg agcagtagct caaaccattt 3300

catatgaagt agcactagga ctaattctcc tctcaacaat tgtatttaca ggaggattta 3360

ccttaactat gtttagcgta acgcaagaaa gcatttgact cctagcccca gcctggcccc 3420

tagcagcaat atgatttgtc tctacactag ccgaaacaaa ccgagccccc tttgacctaa 3480

ccgaaggaga atcagaatta gtctccggtt ttaacgtaga atacgcaggg ggcccattcg 3540

ctctattctt tcttgcagaa tacgctaaca tcttatttat aaatacacta tcagccattt 3600

tatttatagg agccaccaac ttaccaatat taccagaact aactaccatt aatattataa 3660

ttaaagctgc cctactgtcc gctttatttt tatgagtacg agcctcatat ccacgatttc 3720

gatacgacca acttatacat cttgtatgaa aaaatttcct accacttaca ttagcactta 3780

ttatatgaca caccgccatc cctttagcca ctgcaggtct tccccctcag ctataaagga 3840

actgtgcctg aacgcctaag ggacactttg atagagtgaa ccacaggggt taaactcccc 3900

tcagctcctt agaaacaagg gaattgaacc cttcccacag agatcaaaac tctgagtgct 3960

tcctctacac cactttctag taaggtcagc taaataagct tttgggccca taccccaaac 4020

atgttggtta aaacccttcc catactaatg aacccccacg tactcagcat tctgctcatt 4080

agcctaggac ttggaaccac cctcactttt ataagctccc actgactact agcatgaata 4140

ggcctagaaa ttaatactct tgccattatt ccactaatag cccaaaaaca ccatccacga 4200

gcagtagaag ccaccaccaa atactttctt attcaagcaa cagcagcagc cataatccta 4260

tttgccgctt cttccaacgc atgaatttgt ggacaatgag acattaacca aatatcacac 4320

ccaattacct caaccattac tataacagcc ctagcactaa aaattggtct agcccccctc 4380

cacctttgac tcccagaagt ccttcaaggg atcaccttaa caacaggaat aattttatca 4440

acctgacaaa aactagcccc actagccctc attattcaga cagccaacaa tacccatcca 4500

tttctactca cagcactagc attatgctca accctagttg gaggctgagg aggcttaaat 4560

caaacccaac tacgaaaaat tctagcctac tcatctattg cccacctagg atgaataatc 4620

cttgtttccc aaatagcccc tcaaataact cttattgctc taattacata tattgtaata 4680

accaccgcag ctttccttat gttaaacaac attaactcaa caaaaatcat caccctggcc 4740

tccgcttgaa ccaaagcccc cacactcaca gcactcacct gtttagtaat gttatcacta 4800

ggaggactcc cccctctaac aggatttata cctaaatgac ttattattca agaactagcc 4860

aatcaaggac ttgctaccac agcaactatt attgcactta ccgctcttct cagcctttac 4920

ttctacctcc gactaactca cgccctgacc ctcaccctat ccccccaaat caccaacgcc 4980

accaccccct gacgcacccc taacaaacaa cccaccctcg cactatcttt attcactatt 5040

atggctacct gcctactacc tatcacccca acaattttca ttctaatgac ctaggaactt 5100

aggatagcat taagaccatg agccttcaaa gctccaagca ggagtgaaaa cctcctagtt 5160

cctgataaga cttgcaggat tttatcccac atcacctgaa tgcaactcag acacttttat 5220

taagctaaag cctttctagg caggaaggcc tcgatcctac aaactcttag ttaacagcta 5280

agcgctccaa ccagcgagca tccgcctact ttccccgccg tatccgggaa aggcggggaa 5340

agccccggca ggcgttagcc tacgtcttcg ggtttgcaac ccgacatgaa cttcaccaca 5400

gacttggtag aaagaggact caaacctctg taatcggagc tacaatccgc cgcctaaact 5460

ctcggccaac ctacctgtgg caatcacacg ttgatttttt tcaactaatc ataaagacat 5520

tggtaccctt tatttagtat tcggtgcctg agcaggaata gtaggaacag cattaagcct 5580

cttgattcga gcagaactca gccaacccgg ggccctctta ggagacgatc aaatctacaa 5640

cgttattgtt actgcccacg catttgtaat aattttcttt atagtcatac cggttataat 5700

cggcggcttc ggaaattgac tagtccctct aatacttgga gcacctgaca tagcattccc 5760

ccgaataaac aatatgagct tttgactcct gcccccctca tttctccttc ttttagcctc 5820

atctggggta gaagcagggg caggaacagg atgaacagtc tacccaccct tggcaggaaa 5880

cttggctcac gcaggggcct ctgtagacct aacaatcttt tcacttcacc tggccggaat 5940

ctcatctatt ctaggggcta ttaattttat cacaacaact attaatataa aaccacctgc 6000

aatttcacaa taccaaacac ctttatttgt ctgagctgta ttaattacgg cagtactttt 6060

acttctgtcc ctcccagttc tagctgccgg aatcacaatg ctccttacag accgaaacct 6120

aaatactact ttcttcgacc ctgcaggagg aggcgacccc attctttacc aacacttatt 6180

ctggttcttc ggacacccag aagtatatat tttaattctc ccaggatttg gaattatttc 6240

ccacatcgta gcctattatg caggaaaaaa agaacctttc ggttacatag gaatagtatg 6300

agccatgata gccatcgggc tcctaggttt tatcgtctga gcacaccaca tattcacagt 6360

aggaatggac gttgataccc gagcctactt tacttccgct acaataatta ttgccattcc 6420

aacaggtgtt aaagtattta gctgacttgc cactttacac ggcggcgtca tcaaatgaga 6480

gacacctata ctatgggcac taggatttat tttcttgttt acagtgggag gattaacggg 6540

cattgtccta gccaactcat cacttgatat tgttctccac gacacatatt acgtagtagc 6600

acacttccac tatgtcctat ctataggcgc tgtattcgct attgtagcgg gatttgtgca 6660

ctgattcccc ctattcacag ggtacacctt acatagcact tgaacaaaaa ttcacttcgg 6720

aattatattt gttggagtta atttaacttt cttcccccaa catttcctag gtttagcagg 6780

aatgcctcga cgttattctg actacccaga cgcatatact ctttgaaaca cggtatcttc 6840

aatcggatca ttaatctctc ttgtggctgt aattatgttc ttattcatcc tgtgggaagc 6900

attcgcagcc aaacgagaag tttcgtcggt agaactaact ataacaaatg tagagtgact 6960

gcatggctgc cctcctccat accacacctt tgaggagcca gcttttgtac aagtacaagc 7020

aaaataacga gaaagggagg aatcgaaccc ccgtgagatg gtttcaagcc agctgcatgg 7080

ccactctgcc actttcttaa ataagacact agtaaaacca ttaccttgcc ttgtcgaggc 7140

aaaattgtgg gttaaacccc cgcgtgcctt agctcagagc ttaatggcac atccctcaca 7200

actaggattg caagacgcgg cctcccctgt tatagaagaa ctcttacact tccacgacca 7260

cgccctaatg atcgttttac tgattagcgt actggtcctt tacattattg tctcaatggt 7320

gtcaaccaaa ctcaccgata aatatatcct agattcgcaa gaaatcgaaa tcgtatgaac 7380

tatcctacca gccgtcatcc taattatgat cgcactccct tctctgcgga ttctttatct 7440

catggacgag attaatgacc cacacttaac aattaaagcc atgggccacc aatgatactg 7500

aagttatgaa tacacagact acgaagacct aggctttgac tcatacataa tccctactca 7560

agaccttaca ccaggacaat tccggctcct agaaacagac catcgcatgg tagtaccaat 7620

agagtccccc gtccgtgttt tagtctcggc tgaagatgtt ctccactcct gagctgtacc 7680

agccttaggc gtaaaaatag acgcagttcc cggacgccta aaccaaacag cctttattac 7740

ttctcgccct ggcgtattct atggacaatg ctctgaaatc tgcggtgcaa accacagttt 7800

tatgccaatt gttgtagaag ccgtcccttt agaacatttt gaaaactgat catcacttat 7860

aattgaagac gcctcactag gaagctaaaa aggtctagcg tcagcctttt aagctgaagc 7920

ttggtgactc cgccccacct ctagtgacat gcctcaatta aaccccgctc cttgatttat 7980

aatcctcctt ctctcatgag taacttttct aattattctt cccccaaaag ttttagccca 8040

cgaatttagc aacgaaccta ctattatagg ggccgaaaaa tctaaacctg aatcttgaaa 8100

ctgaccatga tactaagctt ttttgaccaa tttataagcc ctacctacat aggaattcca 8160

ctcattgcac tagcaattac cctcccatga atcctttatc caactcctac ctcacgatga 8220

atcaacaacc gcctacttac ccttcaaggt tgatttatca atcgactaag ccagcagatt 8280

ttcttgccta ttaaccaagg gggccataaa tgagccgtcc tactaacatc tctaataatt 8340

ttccttatta cccttaatat gctaggccta ttaccctaca catttacgcc aacaacccaa 8400

ctttctctta atatagcatt cgcagtacca ttatgattgg caactgttat tattggtata 8460

cgtaatcaac caactgcagc actgggacac ctgctcccag aagggacacc gacccctctt 8520

atcccagtac tgatcattat cgaaaccatt agcctgttta ttcgccctct tgcactaggt 8580

gtccggctaa ctgctaacct cactgcaggc cacttactaa ttcaactaat cgcaacagca 8640

gcatttgttc ttctcccact tatgcctact gttgcaatcc taactgccac cgtactattc 8700

cttttaactt tattagaagt tgctgtagca ataatccaag catacgtatt tgtcttactt 8760

ctaagccttt acctacaaga aaacgtctaa tggcccacca agcacacgcg ttccatatag 8820

tagaccccag cccttgaccc ctaaccggcg cagtcggcgc ccttttacta acatctggta 8880

cagcaatctg atttcacttt aattcaatta cccttgcaac cctaggatta attttaacaa 8940

ttttaaccat atatcaatga tggcgagata ttatccggga aggaacattt caaggtcacc 9000

acaccccgcc tgtccaaaaa ggccttcgat acggaataat tctctttatt acatcagaag 9060

tgttcttctt tgcaggattc ttctgagcat tctaccactc cagcctcgca ccaactcctg 9120

aactaggagg ctgctgaccc cccacaggta ttacaacttt agaccctttt gaagtcccac 9180

ttctaaatac agcggttctt ctagcatcag gtgtgaccgt tacatgggcc caccatagct 9240

taatagaagg cgatcgaaaa caggcaattc aatcccttac actaacaatt ctcctaggat 9300

tttactttac cttcctccaa ggcatagaat attatgaagc cccatttact atcgcagatg 9360

gagtttacgg atcaacattt ttcgtcgcca caggattcca cggcctccat gtaatcatcg 9420

gctcaacatt cttagctgtc tgccttctac gccaagtact ttaccatttt acctcaagcc 9480

accatttcgg ctttgaagcc gctgcctgat actgacactt cgtagacgta gtatgactat 9540

tcctatacgt ctcaatttac tgatgaggat cataatcttt ttagtataaa gacaatacaa 9600

atggcttcca accatttaat cttagttaaa atctaagaaa agataatgaa cctaattaca 9660

acagttctat taatctcaac tactttatca ttagtattaa tagcagtttc tttctgactt 9720

ccccaaataa atccagatgc agaaaaactc tccccttacg aatgcgggtt cgacccctta 9780

ggctccgccc gccttccatt ttcaatacga ttctttttag tggctatcct cttcctctta 9840

tttgatctag aaatcgccct actcctcccc cttccatgag gaaatcaact tcaagaacca 9900

aaaaccacct taatctgggc cattttagta atcggcttac taacactggg gttaatctat 9960

gaatgacttc aaggaggctt agagtgggcc gaatagagga ttagtccaat aaagacttct 10020

aatttcggct tagacaatta tggtgaaact ccataatcct cttatgaccc caacacactt 10080

cagctttacc acagcattca ttctaggcct tatgggggtg gcattccacc gaacccattt 10140

attatccgcc cttctctgct tggagggaat aatactctca ctttttattg ccctctcagt 10200

ctgatcttta cagataggtg taacaaattt caccccagca ccaataatat tactagcctt 10260

ctcagcctgt gaagccagcg caggactagc ccttctagta gctacagccc ggactcacgg 10320

cacagaccgc ctacaaaatc taaacttatt acaatgctaa aaatcctcat cccaactctc 10380

atactattcc ctacaatttg actaacccct aaaaaatgat tatgaagctc tgcaacatct 10440

catagcctca ttattgcttt actaagcttt aactgattaa actgagcctc ggaaacagca 10500

tgaagcactt caaacaacta catggcaatt gaccccctat cttcgcctct tctagttctc 10560

acatgttgac tactcccctt aataatttta gccagtcaaa accacaccca agccgaacca 10620

atttcacgac aacgaactta catcagctta ttagcctcgc tacaaacctt tcttatttta 10680

gcattcgggg ctacagaaat cattatgttt tatattatat ttgaagcaac attagtacct 10740

actttaatta ttattactcg ctgagggaac caagcagaac ggctaaacgc aggcacatac 10800

tttttattct acaccctcgc aggatccttg cccctactag ttgctctcct cgccctacaa 10860

acaacaacag ggggactatc aataattaca ctcaacttca gccaacccct ctccttaatt 10920

tcatgaggtg acaagttatg atgaactgcc tgtcttctag cttttctagt caaaatgccc 10980

ttatacggag tccatctctg gcttcctaaa gcacatgtag aagcaccaat tgccggatct 11040

atagtactag ctgcagtact cctcaagtta ggaggctatg gtataatccg catcaccccc 11100

atcctagacc ctctcacaaa agagatagct tatccattta ttattcttgc tctttgagga 11160

atcgtgataa caggaacaat ttgcttacgc caaacagacc ttaaatcgtt aatcgcttac 11220

tcatccgtaa gccacatagg attagtagca agcggcattc taacccaaac accatgaggt 11280

ataacagggg ccatcatcct tataattgct cacggactta catcttccgc attattctgt 11340

ttagcaaaca caagctacga acgaacacac agccgaacta tagcacttgc acgaggaata 11400

caaatattat ttccactaac agcaacctga tgatttattg ccaacctggc taacttagcc 11460

ctgccccccc ttcctaattt aataggagaa ataataatta ttacaacaat atttaattga 11520

tctccctgat caattgtact caccgggctg ggcacattaa tcaccgcaag ctattctctt 11580

tacatatttc tcataaccca acgaggccaa acccccacac acatcaccgc actacctcct 11640

taccacaccc gagaacacct tttaatttca cttcacttaa tccccattat tcttctcacc 11700

attaaaccag aacttatatg agggtgattc tactgcagac gtagtttact aaaaacgttg 11760

gattgtgatt ccaaaaaagg gggttaaatt cccctcgtcc gccggagaga ggcttgcggc 11820

agcagagact gctaatctct tccaccacag ttaaaatctg tggctccctc ggctcctgaa 11880

ggataacagt catccgctgg tcttaggaac caaaaactct tggtgcaaat ccaagcaaga 11940

gctatgcaaa ctacacttgt tcttacttcc tcactaactc taatcttttt cctattggct 12000

tatccactct tcacaacaat aagcccaact cctgtaaaag aagactgagc aattacacat 12060

gtgaaaaccg cagttagcac tgccttcata attagccttt taccagcatt catttttata 12120

gaccaaggcc tcgaagtaat cgtaacaaac tgacactgaa taaatacatc aacattcaat 12180

attaatatca gcctaaaatt tgattactat tcaattactt tcacacctat tgccctatat 12240

gtaacctgat caattctaga atttgctgcg tgatacatac acgaagaccc ctacataaac 12300

cgtttcttta aatacctact cactttcctc attgcaataa ttattctagt cacagccaac 12360

aacatatttc aactatttat tggttgagag ggcgtaggaa tcatatcctt cctgcttatt 12420

ggatgatgat acggccgaac cgatgccaac accgccgccc tccaagcagt aatttataac 12480

cgagttggag atatcggctt aattataact atagcatgat ttgctactaa aattaattca 12540

tgagaaattc aacagatctt ttctttatcc cacaacttta acacaaccct acccgccctg 12600

ggcctaatca ttgccgcaac aggtaaatca gcacaatttg gacttcaccc atgactccct 12660

tccgcgatag aaggccctac cccagtatcc gccctactac attcaagtac gatagtcgta 12720

gctggaattt ttctccttat ccgcctccac ccatttatag aatctaacaa aactattctc 12780

accatttgtc tttgcttagg agcactaacc acgctattta cagctacatg cgctctaacc 12840

caaaacgata ttaaaaaaat tgtagcattc tcaacttcca gccaacttgg acttataata 12900

gttacaatcg gcctcaacca gccccaattg gccttcctac atatttgcac acatgcattc 12960

tttaaagcaa tactattctt atgctcaggg tctattatcc acagcctcaa cgatgaacaa 13020

gatatccgaa aaataggagg actccacaac ctagcaccat ttacctcaac atgtataaca 13080

attggaagcc tagcccttac aggaactccc ttcctagccg gatttttctc caaagacgca 13140

atcattgaag ccttaaatac ctcacatctc aacgcctgag cccttattct tactttaatc 13200

gcaacttcct tcacagccgc ctatagccta cgagttattt tctttgtatc cataggcacc 13260

ccccgattcc tcccattatc ccccattaac gaaaacgacc caaaagtaat taaccctatt 13320

aaacgactcg cctgaggaag tatcattgca ggtctcattc ttacatcaaa cataacaccg 13380

attaataccc ctattatgac aatacctcct ctactaaaac taggagccct tattgttaca 13440

ctagtcggct tatttacagc catagagcta gccaacataa cctcaaaaca actaaaaaca 13500

accccaaata ttaaactaca taatttttca aatatgctag gttattttcc agccattatc 13560

caccgcctaa caccaaaaat cagtcttatt tttggacaaa cactagccaa ccagctagct 13620

gaccaaacat gatttgaagc atcaggtccc aaaggggttt catcaataca actgaaaata 13680

tccgcactca caagtgacac gcagcaagga ataattaaaa catacctaac cacattcctt 13740

atcacactcg cactagcgac acttttagcc ctaatttaaa tagcccgcaa agcccctcga 13800

ctggcccccc gaacaagctc taataccaca aacaagctta acaacagcac cgaggcacaa 13860

ataactaata gacccccacc taaagaatat atcacaccaa caccccctgt gtcctcacat 13920

aatacaaaaa actctttaca aacacttacc acgggtaata aatagttata tcattcacta 13980

ctaaaatatg ccacagcaac ccctactccc actaagtaaa agactacata ctccaatact 14040

gaaacatccc atcacccctc aggatggggc tcagccgcta aagcggctga ataagcaaat 14100

acaactaaca tcccgcctaa ataaattaag aaaagcacga ctgccaaaaa agaagcccca 14160

gaccctgcca aaacagcaca cccagccccg gcaactacaa ctaaaccgag tgctgcaaaa 14220

taaggcgccg gatttgacgc caccgcaacc aaacctaaaa tcaataatac taatcatata 14280

caaaaaagat aagacataaa tttttacctg gattttaacc aagactaatg acttgaaaaa 14340

ccaccgttgt gattcaacta taaaaactcc taatggcaag cctacgaaaa acccaccccc 14400

tacttaaaat tgctaacgac gcggtcgttg acctaccagc cccatcaaat atctcagtat 14460

gatgaaattt cggatcactt ctaggtctct gtttagcaac acaaatcctt acaggactat 14520

ttctcgccat gcactacaca tctgacattg ccactgcatt ctcatcagta gcccatatct 14580

gccgagatgt aaactatgga tgactaatcc gaaacataca tgcaaatgga gcatccttct 14640

tttttatttg tatctatgct cacattgccc gaggattata ctacgggtcc tatctttaca 14700

tagagacatg aaatatcggt gtagtacttc ttcttttagt aataataacc gcctttgtcg 14760

gttacgttct accttgagga caaatatcct tctgaggtgc aactgtcatc acaaatcttt 14820

tatccgcatt cccatacgta ggaaacgaac tagtccaatg aatctgaggc ggattctccg 14880

tagacaacgc taccctaacc cgattctttg ctttccactt cttattcccg tttgtcatcg 14940

caggggctac aattcttcac cttttattcc ttcacgaaac tggctcaaac aacccagcag 15000

gattaaactc tgactcggac aaaatcgctt tccaccctta tttctcttac aaagacctac 15060

taggatttgc agttatacta ttagccctaa cctcattagc cctgttctct cctaatcttt 15120

taggagaccc agacaatttt accccagcca acccgctagt aacccctcct cacattaaac 15180

cagaatgata cttcctcttt gcctacgcca ttctacgatc aatccccaat aaactaggag 15240

gagtattagc actattattt tcaatccttg tccttatagt ggtcccaatt cttcatacct 15300

ccaaacaacg gggcctaacc ttccgacccc tcactcaatt tcttttctga actctagtcg 15360

cagacgtaat tatcttaaca tgaattggag gaatgcctgt tgaacaccct ttcatcatta 15420

ttggtcaaat cgcatccgtg ctttacttca ccctattttt agtactaagc cccctagcag 15480

gctgactaga aaacaaagcg ctcaactgaa actgccctag tagcttaaat ttaaagcgcc 15540

ggtcttgtaa tccggaaacg aaggttaagt cctccctaag gcccaaggag agagaattaa 15600

actcccacca ctagctccca aagctagcat tctgaaatta aactatcctc tgatactaca 15660

gttttatata aacaaacctg aaaaatcaca tacataaata tgtaagcata gacactattg 15720

tcacaccacc acgccttaat caaaaacatg tcaaaactta attgcgtcgc ctacttaaat 15780

tataggttta agtatccatt atgcatatat aagtattaag gtaaacttca ttcagtaaat 15840

ataatgtata acacaacatt gcatgtgctc tgtatatact atgcattata ttacatatat 15900

tatggtatag tacatactat gcattatatt acatatatta tggtatagta catactatag 15960

catcatttgc aatagtatgg ttagtccata ctatgcatta tattacatat ctaggtatag 16020

tacatactag catatcatac atatatatag tatagtacat actatgcata tattacatat 16080

attatggttt agtacatact atgcaatata tacatatact atggtatagt acatatatgc 16140

atcatattac atataatatg gtatagtaca tactatgcat tatattacat atattatggt 16200

atagtacata ctatgcatta tattacatat attatgatat agtacatact atgcattata 16260

ttacatatat tatggtatag tacatactat gcattatatt acataatata tggtactaac 16320

tacacctgat atccttatta aaatgaatct actttcatta aaaagacaac cataaaaatg 16380

tttaaaaact taacaagtaa gaataaaact aagatataca taaaccacaa acatggaaat 16440

tcagaaacca acctaaaaca cctaagaaat agaataatcc ccattacttc tttcgaacgt 16500

tttccatgca aggactcaac attactcggt attcccttat ttaatgtagt aagagaccac 16560

caaccagtat aattaagcgc atatcatgaa tgataagatc agggacaact attgtggggg 16620

tcgcacaaaa tgaactattc ctggcatctg gttcctactt cagggcccca cttcccttga 16680

tcccccctgc aagagcgttt gcgcatatat taatgtggag tcctaattat cctttacccc 16740

cgatgaggag cgttcactta gtaggcatct gatattttat ccggggttac tttcactttg 16800

catttgacta actccttcat aatgcaaact tcttaaggtt gaacattttc cttgcctggc 16860

aaattaaata tccaatactt catcaacatt gatagaagac ttacataact gatatcaagt 16920

gcataaggtt ttattcctta ctccacaata ccctattata gtgccccccc tgcctacgaa 16980

aattaacttt tttcgcgcgt ataaaccccg ttacccccta cgacccagac aagtctattt 17040

ttatctgtca aaccccgaaa ccaggaaaga ccggactagt gtcgtctagc gagtttcgtt 17100

tatgtgctag tcttatagtg ttgcaaaatg caatttcgtt ta 17142

Claims (7)

1. A kind of 1. clupeoid chondrioid genom sequence universal primer, it is characterised in that the Clupoidea fish line grain Body genom sequence universal primer includes ten pair of primers：Wherein,

   Pair of primers：

   Forward primer 1 is as shown in SEQ ID NO.1：5’ - GCAYAAAGGTTTGGTCCT - 3’；

   Reverse primer 1 is as shown in SEQ ID NO.2：5’ - TCGTTGAACAAACGAACCCTT - 3’；

   Second pair of primer：

   Forward primer 2 is as shown in SEQ ID NO.3：5’ - GTTTACGACCTCGATGTTGG - 3’；

   Reverse primer 2 is as shown in SEQ ID NO.4：5’ - AGTGTCTGRGTTGCATYCAG - 3’；

   3rd pair of primer：

   Forward primer 3 is as shown in SEQ ID NO.5：5’ - AGACCATGAGCCTTCAAAG - 3’；

   Reverse primer 3 is as shown in SEQ ID NO.6：5’ - TARCTTCARTATCATTGRTGKCC - 3’；

   4th pair of primer：

   Forward primer 4 is as shown in SEQ ID NO.7：5’ - CCACTCTGMCACTTTCTT - 3’；

   Reverse primer 4 is as shown in SEQ ID NO.8：5’ - TGTGCTTGGTGGGCCATT - 3’；

   5th pair of primer：

   Forward primer 5 is as shown in SEQ ID NO.9：5’ - ATTCAACTAATYGCHACAGC - 3’；

   Reverse primer 5 is as shown in SEQ ID NO.10：5’ - GANAGNGGGTCAATBGCYATRTA - 3’；

   6th pair of primer：

   Forward primer 6 is as shown in SEQ ID NO.11：5’ - CTACAATGCTAAAARTYCTHATCCC - 3’；

   Reverse primer 6 is as shown in SEQ ID NO.12：5’ - TAGCCYHTGCTTGGATTTGCACCAAGAGT - 3’；

   7th pair of primer：

   Forward primer 7 is as shown in SEQ ID NO.13：5’ - GAAGGATAAYAGTCATCCGYTGGTCTTAGG - 3’；

   Reverse primer 7 is as shown in SEQ ID NO.14：5’ - GADGTDGAGAATGCTACRAT - 3’；

   8th pair of primer：

   Forward primer 8 is as shown in SEQ ID NO.15：5’ - GTYACAGCYAACAAYATRTTYCARYT - 3’；

   Reverse primer 8 is as shown in SEQ ID NO.16：5’ - CCAAAGTTTCATCADACTGA - 3’；

   9th pair of primer：

   Forward primer 9 is as shown in SEQ ID NO.17：5’ - CCRAGACTAATGACTTGAAA - 3’；

   Reverse primer 9 is as shown in SEQ ID NO.18：5’ - ATAGTTTAANTTBAGAATNCTAGCTTTGGG - 3’；

   Tenth pair of primer：

   Forward primer 10 is as shown in SEQ ID NO.19：5’ - TCGAGCACCCCTTTATTATTATTGG - 3’；

   Reverse primer 10 is as shown in SEQ ID NO.20：5’ - TAGTACTCCACCATTAACTTATGCG - 3’；

   Tenth pair of primers：

   Forward primer 11 is as shown in SEQ ID NO.21：5’ - CATATCATGAATGATAAGATCA - 3’；

   Reverse primer 11 is as shown in SEQ ID NO.22：5’ - TCTCCATAACCACTCTTTACGC - 3’.
2. 2. a kind of design method of clupeoid chondrioid genom sequence universal primer according to claim 1, It is characterized in that, there is the overlapping region of 45~55bp between the adjacent primer extension product of described ten a pair of of universal primers.
3. 3. a kind of design method of clupeoid chondrioid genom sequence universal primer as claimed in claim 1, its It is characterized in that, all clupeoid chondrioid genom sequences announced at present is downloaded from Genbank databases, Multiple alignment is carried out to these sequences, the region more larger than more conservative region and variation is found out, ten is designed in conservative region A pair of of universal primer.
4. 4. a kind of amplification method of clupeoid chondrioid genom sequence universal primer as claimed in claim 1, its It is characterized in that, the amplification method of the clupeoid chondrioid genom sequence universal primer comprises the following steps：

   （1）DNA extraction agent boxes method extracts the STb gene of Clupoidea fish；

   （2）Using the clupeoid chondrioid genom sequence universal primer group described in claim 1；

   （3）Using Clupoidea fish total genomic dna as masterplate, PCR amplification is carried out；

   （4）1.0% agarose gel electrophoresis of amplimer, RNA isolation kit are tapped and recovered amplified fragments, sequencing and sequence assembly.
5. 5. a kind of amplification method of clupeoid chondrioid genom sequence universal primer according to claim 4, It is characterized in that, step（1）The Genome DNA extraction of the Clupoidea fish, to avoid intestinal contents from polluting, casts out an abdomen Portion, only extracts DNA from its afterbody.
6. 6. a kind of amplification method of clupeoid chondrioid genom sequence universal primer according to claim 4, It is characterized in that, step（3）The reaction system of the PCR amplification forms：DNTP2 μ L of 2.5mM, 10 × Taq DNA polymerase Buffer2.5 μ L, respectively take each forward primer and each reverse primer, 0.2 μ of Taq DNA polymerase of 5U/ μ L that 1 μ L concentration is 10 μM L, the 1 μ L of DNA masterplates solution of 100g/ μ L and 25 μ L are added to sterilizing distilled water；

   The PCR amplification condition is：95 DEG C of pre-degenerations 5min, subsequent 95 DEG C of denaturation 30s, 56 DEG C of annealing 30s, 72 DEG C of extensions 1~4 Min, carries out 35 circulations altogether, finally carries out 72 DEG C and extends 10min, is preserved under the conditions of 4 DEG C.
7. 7. a kind of amplification method of clupeoid chondrioid genom sequence universal primer according to claim 4, It is characterized in that, the step（4）Middle gel electrophoresis uses 1.0% agarose gel electrophoresis, be tapped and recovered sequencing after each sequence according to Secondary head and the tail splicing can obtain mitochondrial genome complete sequence.