KR20130078265A - Infectious cdna clones of foot-and-mouth disease virus of type o and the complete sequences of the clones - Google Patents

Infectious cdna clones of foot-and-mouth disease virus of type o and the complete sequences of the clones Download PDF

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KR20130078265A
KR20130078265A KR1020110147110A KR20110147110A KR20130078265A KR 20130078265 A KR20130078265 A KR 20130078265A KR 1020110147110 A KR1020110147110 A KR 1020110147110A KR 20110147110 A KR20110147110 A KR 20110147110A KR 20130078265 A KR20130078265 A KR 20130078265A
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이광녕
김수미
박종현
고영준
이향심
조인수
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대한민국(관리부서 : 농림축산식품부 농림축산검역본부)
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Abstract

PURPOSE: A cDNA clone is provided to acquire basic knowledge related to replication, expression, and infection of foot-and-mouth disease virus, and to be used vaccine and diagnostic materials, thereby resolving problems of existing inactivated vaccines. CONSTITUTION: A cDNA clone is prepared using foot-and-mouth disease virus with a base sequence of sequence number 1. The clone is prepared using foot-and-mouth disease virus with a base sequence selected from a group consisted of base sequences of sequence numbers 2, 3, 4, and 5. The clone identifies replication and expression difference of a replicon by manipulating untranslated region (UTR) or T7 promoter site of the clone with a base sequence of sequence number 5. In the clone, G is added at 644nt G and C is substituted with T at 9602nt in T7 promoter. A replicon clone has a base sequence of sequence number 6. [Reference numerals] (AA,BB,CC) Foot-and-mouth disease; (DD) Prepare linear DNA (Nsi cut); (EE) Confirm RNA quantification & integrity; (FF) Confirm virus production; (HH) Acquire infectious foot-and-mouse disease virus or express a non-infectious foot-and-mouth virus-like particle and protein

Description

감염력이 있는 구제역바이러스 Ｏ형 ｃＤＮＡ 클론 및 클론의 전체염기서열{Infectious cDNA clones of Foot-and-mouth disease virus of type O and the complete sequences of the clones}Infectious cDNA clones of Foot-and-mouth disease virus of type O and the complete sequences of the clones}

본 발명은 RNA 바이러스 연구를 위한 reverse genetics의 근간이 되는 감염성 클론의 제작 및 활용에 관한 것으로서, 최근 이 분야는 활발히 연구가 진행되고 있다. 특히, 이러한 감염성 클론은 특정 부위에 돌연변이를 인위적으로 가할 수 있는 기법(site-directed mutagenesis)을 병용하면서 RNA 바이러스의 증식, 병원성 등에 대한 이해를 심화시켰고 의료 및 보건 산업분야에도 활용될 수 있는 가능성이 커졌다.
The present invention relates to the fabrication and utilization of infectious clones, which are the basis of reverse genetics for RNA virus research. Recently, this field has been actively studied. In particular, these infectious clones use site-directed mutagenesis in combination with artificial site-directed mutagenesis to deepen their understanding of the proliferation and pathogenicity of RNA viruses, and are likely to be used in the medical and health industries. Got bigger.

이 발명은 구제역바이러스 O형 ME-SA(Middle East-South Asia) 지역형(topotype) 중 세계적으로 그 전파범위가 넓고, 2000년 및 2002년 국내에서도 발생한 Pan Asian strain에 대한 cDNA 클론을 제작하여, linearized DNA 혹은 in vitro transcribed RNA 형태로 특정 세포에 형질도입(transfection)시켰을 경우에 감염력이 있는 구제역바이러스를 얻고자 한 것이다.The present invention has a wide range of global spread of foot-and-mouth virus O type ME-SA (ME-SA) topotype, and produced a cDNA clone for Pan Asian strain which occurred in Korea in 2000 and 2002, The aim was to obtain infectious foot-and-mouth virus when transfected into specific cells in the form of linearized DNA or in vitro transcribed RNA.

구제역바이러스의 감염성 클론은 Zibert 등(1990)에 의해 O1형의 바이러스에 대해 최초로 제작되었는데, 바이러스의 앞쪽 게놈 영역중 단백질을 코딩하지 않는 부위에 존재하고 있는 poly(C) tract의 길이가 60bp 이상은 되어야 세포에서의 감염력을 나타내는 것으로 보고하였다. 그러나 1993년 Rider 등에 의하면 A12 형에 대한 감염성클론 실험에서 poly(C) tract의 길이가 6개 이상이면 실험실에서 배양한 동물세포에서 감염력을 나타내었고 포유마우스에서는 poly(C) tract가 2개 이상인 클론의 경우 모두 동일한 감염력을 나타냈다고 보고하였다. 이러한 연구결과는 poly(C) tract가 바이러스의 복제에 미치는 영향과 역으로 복제나 계대과정에서 poly(C) tract 길의 변화에 대한 이해가 매우 중요하나 그 역할이나 기작에 대한 이해는 제한적임을 말해주고 있다.
The infectious clone of foot-and-mouth disease virus was first produced by Zibert et al. (1990) for a virus of type O1, but the poly (C) tract of 60 bp or more in the region of the front genome of the virus that does not encode a protein is Only reported as infectivity in cells. However, in 1993, according to Rider et al., In infectious clone experiments for type A 12 , if the length of the poly (C) tract was 6 or more, it showed infectivity in animal cells cultured in the laboratory. All clones reported the same infectivity. These findings indicate that the effect of poly (C) tract on viral replication and, conversely, that the understanding of poly (C) tract pathway changes during replication or passage is very important, but its role or mechanism is limited. Giving.

이하 본 발명과 관련된 종래기술에 대해서 기재한다.Hereinafter, the related art related to the present invention will be described.

첫째, 1990년대 개시된 ‘구제역바이러스 O형 (O1K) 전체 genome을 이용한 감염성 cDNA 클론First, the infectious cDNA clone using the whole foot-and-mouth disease virus type O1 (O1K) genome, disclosed in the 1990s.

(J. of virology, June 1990. 2467-2473)’이 존재한다. Poly(C) tract의 길이가 일정값 이하(25 cytidyl residues)일 때는 감염성을 확인하지 못한 점, 이미 20년 이전에 발생한 바이러스로 제작된 점 등에서 본 발명과 다르다.
(J. of virology, June 1990. 2467-2473). When the length of the poly (C) tract is less than a certain value (25 cytidyl residues), it is different from the present invention in that it has not been confirmed to be infectious or made of a virus that occurred 20 years ago.

둘째, 1990년대 개시된 구제역바이러스 A형 (A12) 전체 genome을 이용한 감염성 cDNA 클론(J. of virology, Sept. 1993. p5139-5145)이 존재한다. 이 발명은 혈청형이 전혀 다른 바이러스를 사용한 점에서 본 발명과 기술적으로 본질이 다르다(이발명의 A형이고, 본 발명은 O형임).Secondly, there is an infectious cDNA clone (J. of virology, Sept. 1993. p5139-5145) using the entire foot-and-mouth virus type A (A 12 ) genome disclosed in the 1990s. This invention differs technically in essence from the present invention in that a virus with a completely different serotype is used (type A of the present invention, and the present invention is type O).

본 발명의 과제는 국내발생 구제역바이러스 게놈(RNA)상의 유전적 정보 일체를 그대로 DNA plasmid (클론) 형태로 전환하여 이 클론으로부터 동일한 유전적 정보를 갖고 있는 감염성이 있는 구제역바이러스를 획득할 뿐만 아니라, 구제역바이러스의 증식성 및 병원성에 영향을 미치는 바이러스 게놈부위(염기서열 혹은 아미노산 위치)에 대한 연구를 수행하고, 그러한 부위에 대한 실험적 결과 일부가 궁극적으로 구제역 백신의 개발에 활용될 수 있는 것을 목표로 하였다.The object of the present invention is not only to obtain infectious foot-and-mouth virus having the same genetic information from the clone by converting all the genetic information on the domestically produced foot-and-mouth virus genome (RNA) into a DNA plasmid (clone) form, Conduct research on viral genomic regions (base sequences or amino acid positions) that affect the proliferative and pathogenicity of foot and mouth virus, and aim to ultimately be able to use some of the experimental results on foot and mouth disease vaccines. It was.

작성된 구제역바이러스 클론(DNA plasmid)으로 부터 세포안 또는 세포밖에서 T7 RNA polymerase 효소의 작용에 의해서, 감염력이 있는 구제역바이러스 입자내에 있는 게놈 서열과 동일하게 또는 거의 유사하게, 세포내에서 증식이 가능하고 또한 감염력이 있는 progeny virus를 생성할 수 있는 구제역바이러스 genomic RNA가 안정적으로 합성될 수 있어야 한다. 또한, 클론에 있는 바이러스의 유전적 정보(염기서열)는 인위적인 방법에 의해서 일부 부위가 의도적으로 또는 무작위적으로 특정한 대체적 서열 또는 공백으로서 치환되어 바이러스의 생물학적, 생화학적 및 물리화학적 특성에 어떠한 변화를 주는 가가 관찰될 수 있다. 특히, 증식성과 항원성 그리고 면역원성 등에 영향을 주는 부위에 대한 조작을 통해서 진단 및 백신개발 분야에 유용하게 활용될 수 있다.
By the action of the T7 RNA polymerase enzyme in or out of cells from the prepared DNA plasmids, it is possible to proliferate intracellularly, similarly or almost identical to the genomic sequence in infectious foot-and-mouth virus particles. Foot-and-mouth virus genomic RNA capable of producing infectious progeny virus must be able to be stably synthesized. In addition, the genetic information (base sequence) of the virus in the clone can be altered by artificial means, intentionally or randomly, as a specific alternative sequence or void, which can cause any changes in the biological, biochemical and physicochemical properties of the virus. State can be observed. In particular, it can be usefully used in the field of diagnosis and vaccine development through manipulation of the site affecting proliferation, antigenicity and immunogenicity.

본 발명이 이루고자 하는 기술적 과제들은 이상에서 언급한 기술적 과제들로 제한되지 않으며, 언급되지 않은 또 다른 기술적 과제들은 본 발명의 기재로부터 당해 분야에서 통상의 지식을 가진 자에게 명확하게 이해될 수 있을 것이다. The technical objects to be achieved by the present invention are not limited to the above-mentioned technical problems, and other technical subjects which are not mentioned can be clearly understood by those skilled in the art from the description of the present invention .

본 발명은 서열번호 1과 같은 염기서열을 갖는 구제역바이러스를 이용하여 작성한 cDNA 클론에 관한 것이다. 상세하게는 서열번호 1에 포함된 구제역바이러스 전체염기서열의 주된 주형(template)이 된 구제역바이러스를 이용하여 작성한 cDNA 클론에 관한 것이다. 즉, 2002년 국내분리 구제역바이러스를 이용하여 작성한 최초의 구제역 cDNA 클론(P11) 및 그 서열에 관한 것이며, 보다 상세히 설명하면 다음과 같다.
The present invention relates to a cDNA clone prepared using foot-and-mouth virus having a nucleotide sequence as shown in SEQ ID NO: 1. Specifically, the present invention relates to a cDNA clone prepared by using a foot-and-mouth virus that has become the main template of the entire foot-and-mouth virus base sequence contained in SEQ ID NO: 1. That is, it relates to the first foot-and-mouth disease cDNA clone (P11) and its sequence created in 2002 using the isolated foot-and-mouth disease virus, which will be described in more detail as follows.

□ P11, 11093 bases□ P11, 11093 bases

* 1-625: pBluescript II SK(+) vector, 1-615* 1-625: pBluescript II SK (+) vector, 1-615

* 625: G(pBluescript II SK(+) vector) -> C(P11)* 625: G (pBluescript II SK (+) vector)-> C (P11)

* 626-644: T7 promotor sequence (TAATACGACTCACTATAGG)* 626-644: T7 promotor sequence (TAATACGACTCACTATAGG)

* 645-8877: 구제역바이러스 서열 (0/SKR/2002)645-8877: foot-and-mouth virus sequence (0 / SKR / 2002)

* 8846-8877: poly(A), 32bps* 8846-8877: poly (A), 32bps

* 8925-11093: pBluescript II SK(+) vector, 793-2961
8925-11093: pBluescript II SK (+) vector, 793-2961

본 발명은 서열번호 2와 같은 염기서열, 서열번호 3와 같은 염기서열, 서열번호 4와 같은 염기서열 및 서열번호 5와 같은 염기서열로 이루어진 군에서 선택된 어느 하나의 염기서열을 갖는 구제역바이러스를 이용하여 작성한 cDNA 클론에 관한 것이다. 즉, P11을 수정하여 2002년 국내발생 소 및 돼지유래 구제역바이러스 서열로 단백질 번역부위 일체를 치환한 클론(ASP, ASC, AS19, AS10) 및 그 서열에 관한 것이며, 보다 상세히 설명하면 다음과 같다.
The present invention uses foot-and-mouth virus having a nucleotide sequence selected from the group consisting of a nucleotide sequence such as SEQ ID NO: 2, a nucleotide sequence such as SEQ ID NO: 3, a nucleotide sequence such as SEQ ID NO: 4, and a nucleotide sequence such as SEQ ID NO: 5 It relates to cDNA clones created by. That is, it relates to a clone (ASP, ASC, AS19, AS10) and its sequence in which P11 was modified to replace all protein translation sites with domestically generated bovine and swine-based foot-and-mouth virus sequences in 2002, which will be described in more detail as follows.

□ ASP, 11093bps□ ASP, 11093bps

* 645-8877: P11에 2002년 구제역감염 돼지유래 임상시료의 구제역바이러스를 세포에 계대한 서열 반영 (반영구간: 1347[TCTAGA, Xba I] to 8083[CTGCAG, Pst I])
* 645-8877: P11 reflects the sequence of the foot-and-mouth virus of the foot-and-mouth disease-derived clinical sample in 2002 in P11 (semi-period: 1347 [TCTAGA, Xba I] to 8083 [CTGCAG, Pst I])

□ ASC, 11093bps□ ASC, 11093bps

* 645-8877: P11에 2002년 구제역감염 소 유래 임상시료의 구제역바이러스를 세포에 계대한 서열 반영 (반영구간: 1347[TCTAGA, Xba I] to 8083[CTGCAG, Pst I])
* 645-8877: P11 reflects the sequence of the foot-and-mouth virus of a foot-and-mouth disease-derived clinical sample from P11 in 2002 (semi-period: 1347 [TCTAGA, Xba I] to 8083 [CTGCAG, Pst I])

□ AS19, 11093bps□ AS19, 11093bps

* 645-8877: P11에 2002년 구제역감염 소 유래 임상시료의 구제역바이러스 서열 반영(반영구간: 1347(TCTAGA, Xba I) to 8083(CTGCAG, Pst I))
* 645-8877: Reflected the foot-and-mouth virus sequence of the foot-and-mouth disease-derived clinical sample in 2002 in P11 (semi-period: 1347 (TCTAGA, Xba I) to 8083 (CTGCAG, Pst I))

□ AS10, 11093bps□ AS10, 11093bps

* 645-8877: P11에 2002년 구제역감염 돼지유래 임상시료의 구제역바이러스 서열 반영(반영구간: 1347(TCTAGA, Xba I) to 8083(CTGCAG, Pst I))
* 645-8877: P11 reflects the foot-and-mouth virus sequence of the foot-and-mouth disease-derived clinical sample in 2002 (semi-period: 1347 (TCTAGA, Xba I) to 8083 (CTGCAG, Pst I))

본 발명은 서열번호 5와 같은 염기서열을 갖는 클론의 UTR(untranslated region) 또는 T7 promotor 부위를 수정하여 replicon의 복제 및 발현의 차이를 확인하게 한 클론으로서, T7 promotor 부위 644nt G 뒤에 G가 하나 더 삽입되고, 9602nt 위치의 C가 T로 치환된 것을 특징으로 하는 클론에 관한 것이다. 즉, AS10의 UTR(untranslated region) 또는 T7 promotor 부위를 수정하여 replicon의 복제 및 발현의 차이를 확인하게 한 클론들(AS10T7G3)에 관한 것이며, 보다 상세히 설명하면 다음과 같다.
The present invention is a clone for modifying the UTR (untranslated region) or T7 promotor region of a clone having a nucleotide sequence as shown in SEQ ID NO: 5 to identify the difference in replication and expression of the replicon, one more G after the T7 promotor region 644nt G A clone is inserted and the C at 9602nt position is substituted with T. That is, it relates to the clones (AS10T7G3) that allow the modification of the replication and expression of replicon by modifying the untranslated region (UTR) or T7 promotor region of AS10.

□ AS10T7G3, 11094bps□ AS10T7G3, 11094bps

* AS10의 T7 promotor 부위 644nt G 뒤에 G가 하나 더 삽입됨(TAATACGACTCACTATAGG -> TAATACGACTCACTATAGGG)* One more G is inserted after 644nt G of the T7 promotor region of AS10 (TAATACGACTCACTATAGG-> TAATACGACTCACTATAGGG)

* C9602T (AS10의 9602nt 위치의 C가 T로 치환됨, pBluescript vector 영역)
* C9602T (C at 9602nt position of AS10 is replaced by T, pBluescript vector region)

본 발명은 서열번호 5와 같은 염기서열을 갖는 클론의 UTR(untranslated region) 또는 T7 promotor 부위를 수정하여 replicon의 복제 및 발현의 차이를 확인하게 한 클론으로서, 1309nt 위치에서 A가 G로 치환되고, 9458nt 위치의 C가 A로 치환된 것을 특징으로 하는 클론에 관한 것이다. 즉, AS10의 UTR(untranslated region) 또는 T7 promotor 부위를 수정하여 replicon의 복제 및 발현의 차이를 확인하게 한 클론들(AS10IR)에 관한 것이며, 보다 상세히 설명하면 다음과 같다.
The present invention is a clone for modifying the UTR (untranslated region) or T7 promotor region of the clone having a nucleotide sequence as shown in SEQ ID NO: 5, to identify the difference in replication and expression of the replicon, A is replaced by G at the 1309nt position, C at 9458 nt is replaced with A. That is, it relates to clones (AS10IR) that allow the modification of the replication and expression of the replicon by modifying the UTR (untranslated region) or T7 promotor region of AS10.

□ AS10IR, 11093bps□ AS10IR, 11093bps

* A1309G (1309nt 위치에서 AS10의 A가 G로 치환됨, 구제역의 IRES 영역)* A1309G (A in AS10 is replaced by G at position 1309nt, IRES region of foot and mouth disease)

* C9458A (AS10의 9458nt 위치의 C가 A로 치환됨, pBluescript vector 영역)
* C9458A (C at 9458nt position of AS10 is substituted with A, pBluescript vector region)

본 발명은 서열번호 5와 같은 염기서열을 갖는 클론의 UTR(untranslated region) 또는 T7 promotor 부위를 수정하여 replicon의 복제 및 발현의 차이를 확인하게 한 클론으로서, 1309nt 위치의 A가 G로 치환되고, 9602nt 위치의 C가 T로 치환된 것을 특징으로 하는 클론.에 관한 것이다. 즉, AS10의 UTR(untranslated region) 또는 T7 promotor 부위를 수정하여 replicon의 복제 및 발현의 차이를 확인하게 한 클론들(AS10T7G3IR)에 관한 것이며, 보다 상세히 설명하면 다음과 같다.
The present invention is a clone for modifying the UTR (untranslated region) or T7 promotor region of the clone having a nucleotide sequence such as SEQ ID NO: 5, the clone is to identify the difference in replication and expression of the replicon, A in the 1309nt position is substituted with G, C at position 9602nt is substituted with T. That is, it relates to the clones (AS10T7G3IR) that allow the modification of the replication and expression of replicon by modifying the UTR (untranslated region) or T7 promotor region of AS10.

* AS10T7G3IR, 11094bps* AS10T7G3IR, 11094bps

AS10의 T7 promotor 부위 644nt G 뒤에 G가 하나 더 삽입됨(TAATACGACTCACTATAGG -> TAATACGACTCACTATAGGG)One more G is inserted after 644nt G of the T7 promotor region of AS10 (TAATACGACTCACTATAGG-> TAATACGACTCACTATAGGG)

A1309G (AS10의 1309nt 위치의 A가 G로 치환됨, 구제역의 IRES 영역)A1309G (A at position 1309nt of AS10 is substituted for G, IRES region of foot and mouth disease)

C9602T (AS10의 9602nt 위치의 C가 T로 치환됨, pBluescript vector 영역)
C9602T (C at 9602nt position of AS10 is replaced by T, pBluescript vector region)

본 발명은 서열번호 6과 같은 염기서열을 갖는 replicon 클론에 관한 것이다. 즉, ASP의 S fragment와 poly(C) 부위를 교체하기 위해 작성한 변형된 ASP클론(ASP_A618G)및 그 서열과 이 클론에서 해당부위가 교체된 replicon 클론(ASPs12L) 및 서열에 관한 것이며, 보다 상세히 설명하면 다음과 같다.
The present invention relates to a replicon clone having a nucleotide sequence as shown in SEQ ID NO: 6. That is, it relates to a modified ASP clone (ASP_A618G) and its sequence and replicon clone (ASPs12L) and sequence in which the corresponding site was replaced to replace the S fragment and the poly (C) region of the ASP. Is as follows.

□ ASP_A618G, 11093bps□ ASP_A618G, 11093bps

* ASP의 618nt A를 G로 치환함. 이로써 클론내 유일한 Asc I 제한효소 인식부위(GGCGCGCC)가 생성되었고, 이 부위와 Xba I 제한효소 인식부위(TCTAGA, 1347-1352) 사이를 돼지유래가검시료의 세포계대주 바이러스를 대상으로 증폭하여 교체함.
* 618nt A in ASP is replaced with G. This generated the only Asc I restriction enzyme recognition site (GGCGCGCC) in the clone, and amplified and replaced between this site and the Xba I restriction enzyme recognition site (TCTAGA, 1347-1352) in the swine-derived cell lineage virus. .

□ ASPs12L, 11083bps□ ASPs12L, 11083bps

* ASP와 비교할 때, A618G로 치환, A732G로 치환 C819T로 치환, T865C로 치환, 1013-1034 poly(C) 부위(CCCCCCTCCCCCCCCCCCCCCC)가 CCCCCCCCCCCC(12개)로 교체됨.
* Compared to ASP, substituted with A618G, substituted with A732G substituted with C819T, substituted with T865C, and 1013-1034 poly (C) site (CCCCCCTCCCCCCCCCCCCCCCCC) replaced with CCCCCCCCCCCC (12).

본 발명은 ASPs12L에서 poly(C) 부위를 12개에서 24~36개로 연장한 감염성 클론에 관한 것이다. 즉, ASPs12L에서 유래된 poly(C) 부위를 연장한 감염성이 확인된 클론(ASPs24L) 및 그 서열에 관한 것이며, 보다 상세히 설명하면 다음과 같다.
The present invention relates to infectious clones extending from 12 to 24 to 36 poly (C) sites in ASPs12L. That is, the present invention relates to an infectious clone (ASPs24L) having its poly (C) site derived from ASPs12L and its sequence, which will be described in more detail as follows.

□ ASPs24L, 11095bps□ ASPs24L, 11095bps

* Poly(C)부위를 12개에서 24개 이상으로 늘림. Capillary type의 sequencer로서 판별할 수 있는 병렬로 연결된 C의 개수에는 제한이 있음. * Increased Poly (C) area from 12 to 24 or more. There is a limit to the number of parallel Cs that can be identified as capillary type sequencers.

* 여기에서는 24개로 기록하나 site-dircted mutation에서 의도한 바대로 변형되었다면 실제로는 36개 정도까지 연장되었을 수 있음.
* It is recorded here as 24, but if it is modified as intended by site-dircted mutation, it may actually be extended to 36.

본 발명은 ASPs24L에서 VP1 34th 부위 아미노산서열을 교체한 감염성 클론에 관한 것이다. 즉, ASPs24L에서 VP1 34th 부위 아미노산서열을 교체한 감염성이 확인된 클론(ASPs11F) 및 그 서열에 관한 것이며, 보다 상세히 설명하면 다음과 같다.
The present invention relates to an infectious clone with the replacement of the VP1 34th site amino acid sequence in ASPs24L. That is, the present invention relates to an infectious clone (ASPs11F) having replaced the VP1 34th site amino acid sequence in ASPs24L and its sequence.

□ ASPs11F, 11092bps□ ASPs11F, 11092bps

* ASPs24L의 4022nt C를 T로 치환함 (758째 아미노산, VP1의 34번째 아미노산이 F(phenylalanine)에서 L(leucine)으로 치환됨)* 4022nt C of ASPs24L is replaced by T

* Mutagenesis 과정에서 poly(C)부위의 C의 갯수가 24개에서 11개로 줄어듦
* The number of C in the poly (C) region was reduced from 24 to 11 during the mutagenesis process.

본 발명은 구제역바이러스 클론의 poly(C) 부위를 연장하여 감염력의 획득력에 변화를 가져올 수 있는 돌연변이 작성법, 이 때 사용된 프라이머 서열에 관한 것이다. 이에 대해 보다 상세히 설명하면 다음과 같다.
The present invention relates to a mutagenesis method which can extend the poly (C) site of foot-and-mouth virus clones, resulting in a change in the acquisition power of infectivity, and the primer sequence used at this time. This will be described in more detail as follows.

□ High fidelity PCR by Phusion Hot Start DNA Polymerase (프라이머는 poly(C)의 길이를 36개 까지 연장할 수 있도록 설계된 Forward primer(5'-CCCCCCCCCCCCCCCCCCAAGTTTTTAC-3')와 Reverse primer( 5'-GGGGGGGGGGGGGGGGTGAAAGGT-3')를 사용하여 ASPs12L를 주형으로 함)□ High fidelity PCR by Phusion Hot Start DNA Polymerase (Primer is designed to extend the length of poly (C) up to 36 Forward primer (5'-CCCCCCCCCCCCCCCCCCAAGTTTTTAC-3 ') and Reverse primer (5'-GGGGGGGGGGGGGGGGTGAAAGGT-3' Using ASPs12L as a template)

□ Dpn I 제한효소를 처리하여 주형으로 상용된 플라스미드를 분해함. (Dpn I(TOYOBO)은 methylated된 기존의 플라스미드만 분해함)□ Decompose the plasmid commercially available as a template by treating Dpn I restriction enzyme. ( Dpn I (TOYOBO) only degrades conventional methylated plasmids)

□ Self-ligation시킨 후에 transformation 시킴 (Electromax E. coli DH10B cells (Invitrogen) 사용)
□ Perform transformation after self-ligation (use Electromax E. coli DH10B cells (Invitrogen))

본 발명은 Poly(C)의 길이가 24개 이상으로 연장되어 감염력이 확인된 구제역바이러스 cDNA 클론이 형질전환된 대장균의 대량 배양법에 관한 것이다. 이에 대해 보다 상세히 설명하면 다음과 같다.
The present invention relates to a mass culture method of Escherichia coli transformed with a foot-and-mouth virus cDNA clone whose poly (C) is extended to 24 or more in length and confirmed its infectivity. This will be described in more detail as follows.

□ 정확한 원인은 알 수 없지만, poly(C)의 길이가 24개 이상으로 연장되어 감염력이 있는 구제역바이러스 cDNA 플라스미드로 최초 확인된 대장균 클론의 경우는 miniprep을 통해 mutagenesis가 성공된 사실은 확인이 가능하였으나 master plate에서 해당 colony가 잘 자라지 않는 경우가 있었고, midiprep을 위해 일반적으로 사용되는 형질전환된 대장균의 배양조건(16-20시간, 250rpm, 37℃)에서는 증식되지 않는 경우가 있었음.
□ The exact cause is unknown, but in the case of E. coli clones first identified as infectious foot-and-mouth virus cDNA plasmids with more than 24 poly (C) lengths, miniprep confirmed that mutagenesis was successful. The colony did not grow well on the master plate, and the colony did not grow under the culture conditions (16-20 hours, 250rpm, 37 ℃) of transformed Escherichia coli commonly used for midiprep.

□ 따라서, 이러한 경우 감염력이 확인된 클론의 플라스미드를 대장균 세포에 재차 형질전환시켜서 약 1ml에 해당하는 배지에서 1시간 정도 배양후 이 배지액 전량을 항생제가 포함된 배지 200ml에 부어준 뒤 100rpm 및 37℃ 온도조건에서 30시간 이상 배양시킴으로서 200ml에서 약 100ug이상의 플라스미드를 획득할 수 있었음.□ Therefore, in this case, the cloned plasmid of confirmed infectivity was transformed into E. coli cells again, and cultured for about 1 hour in a medium corresponding to about 1 ml, and then the whole amount of the medium was poured into 200 ml of the medium containing antibiotics, followed by 100 rpm and 37 By incubating for 30 hours or more at a temperature of ℃ ℃ was able to obtain a plasmid of more than about 100ug in 200ml.

국내분리된 구제역바이러스 O형 PanAsia 형 바이러스의 게놈 정보를 그 조성과 순서에 있어서 99.9% 이상 동일(poly(C) tract 및 poly(A) tract 제외, 이 부위는 공시된 바이러스의 게놈에서도 그 길이를 정확히 파악하기 곤란함)하거나 아미노산 서열에서는 100% 동일하게 DNA형태로 세균의 플라스미드 형태로 제작하여, 이후 구제역바이러스 감수성 세포에서 이러한 클론(플라스미드)의 구제역바이러스 RNA 게놈을 생성해 내고, 복제 및 발현을 일으키고 또한 최종적으로는 공시된 바이러스와 같이 감염력이 있는 바이러스입자를 생산해 내는 것을 다루었다. 다른 혈청형의 바이러스로 감염성클론이 작성된 사례가 있지만 구제역 혈청형 O내의 PanAsia형 바이러스는 최초로 2002년 국내발생바이러스의 세포계대주에 대해 감염성 클론을 제작하였다. More than 99.9% of genome information of Korean isolated foot-and-mouth virus O PanAsia virus is identical in composition and order (except poly (C) tract and poly (A) tract). Difficult to determine exactly) or 100% identical amino acid sequence in the form of a bacterial plasmid in the form of bacteria, and then to produce foot-and-mouth virus RNA genome of these clones (plasmid) in foot-and-mouth virus-sensitive cells, and replication and expression And finally producing infectious viral particles, such as the published virus. Infectious clones were produced with other serotype viruses, but the PanAsia virus in foot-and-mouth disease serotype O was the first to produce infectious clones against the cell lineage of domestically generated viruses in 2002.

본 발명은 특히 감염력이 확인되지 않는 replicon형태의 클론 및 감염력이 확인되는 감염성 클론을 포함하고 있으며, 이들 간의 서열상 차이가 사실상 미미한 점을 고려할 때 감염력에 영향을 주는 인자에 대한 연구를 위한 클론 세트를 제공하고 있으며, 복제에 영향을 주는 인자에 대한 연구결과도 나타내었다. 특히, poly(C)의 길이를 조절하는 방법을 제공하고 바이러스의 생존에 있어서 이들의 역할을 파악할 수 있는 재료와 기초결과를 제공한다. 또한, 대장균의 생장과 구제역 게놈이 들어가 있는 플라스미드와의 상호작용에 대한 단서를 제공한다. 또한, 이들의 전체서열과 클론의 기본골격을 확인할 수 있는 제한효소의 사용과 전체서열을 분석할 수 있는 프라이머의 제공으로 원하는 방식으로 클론을 조작하여 백신 및 진단법 개발, 병원성 관련인자 탐색 연구등에 효과적으로 사용될 수 있는 자료를 제공한다.The present invention specifically includes replicon clones without infectivity and infectious clones with confirmed infectivity, and considering the fact that the differences in sequence among them are virtually insignificant, a clone set for the study of factors affecting infectivity The results of studies on factors that affect replication are also presented. In particular, it provides a method for controlling the length of poly (C) and provides materials and basic results to understand their role in the survival of the virus. It also provides clues to the growth of Escherichia coli and its interaction with the plasmid containing the foot-and-mouth genome. In addition, the use of restriction enzymes that can identify the entire sequence and basic skeleton of these clones, and the provision of primers that can analyze the entire sequence to manipulate the clones in the desired manner to effectively develop vaccines and diagnostics, and to study pathogenic factors. Provide data that can be used.

결과적으로 구제역바이러스의 복제와 발현 그리고 감염과 관련된 기초지식을 획득하는데 이용될 뿐만아니라 일부유전자 및 부위를 조작하여 백신 및 진단의 재료로 활용할 수 있으므로 기존의 구제역 불활화 백신으로 해결할 수 없었던 분야에 도움을 줄 수 있다.As a result, it is not only used to obtain basic knowledge related to the replication and expression of foot-and-mouth virus, and to be used as a material for vaccines and diagnosis by manipulating some genes and sites, thereby helping the field which could not be solved by the existing foot-and-mouth inactivated vaccine. Can give

도 1에서 구제역바이러스의 전체 게놈을 플라스미드에 클로닝한 그림이다.
도 2는 구제역바이러스 전체 게놈이 클로닝될 때 나타나는 제한효소 소화 패턴이다.
도 3은 P11 클론을 수정하기위해 공시된 4종의 국내발생구제역 바이러스와 이들이 클로닝된 그림이다.
도 4는 구제역 전체게놈 클론을 세포에 형질도입시켜 그 감염여부를 확인하는 실험법 소개이다.
도 5는 도4의 4개 클론들이 작성되어 실험결과를 나타낸 것이다.
도 6은 도5에서 발현력이 가장 좋은 AS10을 변형시킨 클론들을 DNA형태로 세포에 도입한 결과이다.
도 7은 도5에서 발현력이 가장 좋은 AS10을 변형시킨 클론들을 RNA형태로 세포에 도입한 결과이다.
도 8은 ASP클론을 감염력을 얻게 하기 위해 변형시킨 클론이다.
도 9는 ASP클론의 감염력이 확인되어 그 상층액에 대한 염기서열분석결과이다.
도 10은 감염력이 있는 구제역 바이러스 획득 또는 비감염력의 구제역 바이러스 유사입자 및 단백질의 발현에 관한 것이다. In Fig. 1, the whole genome of foot-and-mouth virus was cloned into a plasmid.
Figure 2 is a restriction enzyme digestion pattern that appears when the foot and mouth virus whole genome is cloned.
3 shows four domestic foot-and-mouth disease viruses that have been published for the modification of P11 clones and their clones.
Figure 4 is an introduction to the test method for confirming the infection by transducing the whole foot-and-mouth genome clone cells.
5 shows four experimental results of four clones of FIG. 4.
FIG. 6 shows the results of introducing clones modified with AS10 having the best expression ability into cells in the form of DNA.
FIG. 7 shows the results of introducing clones modified with AS10 having the best expression ability into cells in RNA form.
8 is a modified clone of the ASP clone to obtain infectivity.
9 is confirmed the infectivity of the ASP clone is the sequencing results of the supernatant.
10 relates to the expression of infectious foot and mouth virus acquisition or non-infective foot and mouth virus like particles and proteins.

본 발명은, 구제역바이러스 전체게놈 정보가 이중나선의 circular DNA 벡터에 실려서 T7 RNA polymerase의 작용에 의해 구제역바이러스 단백질의 발현이나 구제역바이러스의 RNA 게놈의 복제가 확인되지 않는 형태의 원형의 DNA클론 (P11)을 제공한다. The present invention is a circular DNA clone (P11) in which the expression of the foot-and-mouth virus virus or the replication of the foot-and-mouth virus virus genome is not confirmed by the action of T7 RNA polymerase in the whole genome of foot-and-mouth virus genome information in a double-stranded circular DNA vector. ).

이 클론으로부터 구제역바이러스 전체게놈 정보가 이중나선의 circular DNA 벡터에 실려서 T7 RNA polymerase의 작용에 의해 구제역바이러스 단백질을 합성하고 구제역바이러스의 RNA 게놈의 복제가 확인되지만 구제역 감수성 세포로부터 감염성이 있는 구제역 바이러스 입자를 생산할 수는 없는, 즉 replicon 형태의 RNA 게놈과 그 유래의 구제역단백질 생산이 확인되는, DNA 클론(AS10, ASC 및 ASP) 및 이들 클론의 특정 부위에 돌연변이를 유도하거나 특정부위를 교체한 DNA 클론들(ASPs12L 등)을 제공한다. 이들은 모두 감염성이 있는 구제역 바이러스 입자를 즉각적으로 생산할 수는 없지만 RNA 게놈 복제 및 단백질 번역 효율에 상호간 차이를 나타낸다. From this clone, the full-mouth disease virus genome information is loaded into a double-stranded circular DNA vector to synthesize the foot-and-mouth virus protein by the action of T7 RNA polymerase and to confirm the replication of the RNA genome of the foot-and-mouth virus, but infectious foot-and-mouth virus particles from foot-and-mouth susceptible cells. DNA clones (AS10, ASC, and ASP) that are unable to produce RNA genomes of replicon form and foot-and-mouth protein derived from them, and DNA clones that induce mutations or replace specific sites in these clones (Such as ASPs12L). They cannot all produce infectious foot-and-mouth virus particles immediately, but show mutual differences in RNA genome replication and protein translation efficiency.

또한 위 클론중 ASPs12L에서 유래된, 구제역바이러스 전체게놈 정보가 이중나선의 circular DNA 벡터에 실려서 T7 RNA polymerase의 작용에 의해 구제역 감수성 세포로 부터 감염성이 있는 구제역바이러스 입자를 생산할 수 있는 구제역바이러스 RNA 게놈을 세포내 또는 세포밖에서 합성해 낼 수 있는 DNA 클론(ASPs35L 및 ASPs11F)을 제공한다.In addition, the full-mouth disease virus genome information derived from ASPs12L of the above clones was loaded into a double-stranded circular DNA vector, and the action of the T7 RNA polymerase produced a foot-and-mouth virus RNA genome capable of producing infectious foot-and-mouth virus particles from foot-and-mouth susceptible cells. Provided are DNA clones (ASPs35L and ASPs11F) that can be synthesized intracellularly or extracellularly.

또한, DNA 클론(ASPs12L) 클론의 poly(C) tract 길이를 연장하여 새로운 클론(ASPs24L)을 작성한 뒤, 이 클론을 세포에 도입시키는 실험을 하게 되면, 세포내에서 RNA의 복제과정중에 일어나는 무작위적 혹은 선택적 변이에 의해 DNA 클론에서 의도적으로 변화시킨 부위(VP1, 34th aa)가 둘 다 소수성(hydrophobic)한 아미노산인 L(leucine)에서 F(Phenylalanine)로 변화됨을 관찰하였다. 이 때 사용되는 poly(C) 부위를 연장시킬 수 있는 돌연변이에 사용되는 올리고뉴클레오타이드 서열 및 돌연변이 생성법 그리고 poly(C) tract가 최소 24개 이상으로 연장된 DNA 클론(ASPs24L)를 제공한다. In addition, when a new clone (ASPs24L) is made by extending the poly (C) tract length of the DNA clone (ASPs12L) clone and then introducing the clone into the cell, randomization occurs during the replication of RNA in the cell. Alternatively, it was observed that the sites (VP1, 34th aa) intentionally changed in DNA clones by selective mutations changed from L (leucine), F (Phenylalanine), to both hydrophobic amino acids. Provided are oligonucleotide sequences and mutagenesis methods used for mutations that can extend the poly (C) site used, and DNA clones (ASPs24L) with at least 24 poly (C) tracts.

위와 같이 RNA 게놈의 복제 및 구제역바이러스 단백질의 생산을 유도할 수 있지만 감염성이 있는 온전한 구제역바이러스 입자의 존재를 간접적으로 확인할 수는 없었던 DNA 클론(ASPs12L)에서 감염성의 획득에 핵심적인 위치로 발견된 VP1의 34번째 아미노산을 L34F로 돌연변이 유도를 해주면 poly(C) tract 길이를 늘리지 않고도 감염성이 있는 구제역바이러스의 생산을 유도할 수 있는 이 아미노산 부위 및 이 부위가 치환된 DNA 클론(ASPs11F)을 제공한다. 이 때 poly(C)의 길이는 12개에서 11개로 오히려 1개 줄어든다.VP1 was found to be a key position for obtaining infectivity in DNA clones (ASPs12L) that could induce the replication of RNA genomes and the production of foot-and-mouth virus proteins, but indirectly confirmed the presence of infectious intact foot-and-mouth virus particles. Mutation induction of the 34th amino acid into L34F provides this amino acid site and a DNA clone with this site (ASPs11F) that can induce the production of infectious foot and mouth virus without increasing the poly (C) tract length. At this time, the length of poly (C) is reduced from 1 to 12 rather than 12.

구제역 바이러스 RNA 게놈으로 부터 작성된 DNA클론이 감염성이 있는 구제역바이러스를 생산하는 여부를 확인하는 방법을 구체적으로 설명하면;
Specifically, a method for confirming whether a DNA clone prepared from the foot-and-mouth virus RNA genome produces an infectious foot-and-mouth virus is described in detail;

(1) 클로닝된 plasmid 전체서열을 확인하여 구제역 RNA 게놈서열과 일치하는 여부를 확인하는 단계;(1) checking the cloned plasmid entire sequence to determine whether it matches the foot-and-mouth RNA genome sequence;

(2) 클로닝된 plasmid를 대량으로 얻기 위하여 형질전환된 특정 대장균을 배양하고 plasmid를 회수하는 단계;(2) culturing the transformed specific E. coli and recovering the plasmid to obtain a large amount of the cloned plasmid;

(3) 회수된 plasmid를 특정한 제한효소(Nsi I)를 사용하여 구제역바이러스 게놈의 끝단에 해당하는 poly (A)의 뒤쪽을 절단하여 선형의 이중가닥 DNA형태로 만드는 단계;(3) cutting the back of the poly (A) corresponding to the end of the foot and mouth virus genome by using a specific restriction enzyme (Nsi I) to form a linear double-stranded DNA;

(4) 선형 이중나선 구조의 DNA 끝단을 Klenow 효소를 사용하여 Blunt end로 변형시키는 단계;(4) transforming the DNA ends of the linear double helix structure into Blunt ends using Klenow enzyme;

(5) T7 RNA polymerase가 발현되는 세포주(BHK T7-9)에 위 (4)의 DNA를 형질도입(transfection)시켜서 구제역바이러스 게놈 RNA와 (거의) 동일한 서열의 RNA를 세포내에서 합성되도록 하는 단계, 또는 세포밖 튜브내에서 in-vitro transcribed 된 RNA를 형질도입시키는 단계;(5) transducing the DNA of (4) to the cell line (BHK T7-9) expressing T7 RNA polymerase to allow RNA synthesis of (almost) the same sequence as the foot and mouth virus genomic RNA Or transducing in-vitro transcribed RNA in an extracellular tube;

(6) DNA로부터 합성된 구제역바이러스 유사 RNA 게놈이 세포내에서 증식하고 단백질을 번역하는 단계;(6) foot-and-mouth virus-like RNA genomes synthesized from DNA proliferate intracellularly and translate proteins;

(7) 세포내에서 일반적인 구제역바이러스의 복제와 차이점을 찾기 어려운 방식으로 복제된 (염기서열이 구제역바이러스와 (거의) 일치하는) RNA 게놈과 동시에 세포내에서 발현된 구제역 입자 단백질이 조합되어 바이러스입자로 완성된 후 이 바이러스가 또 같은 또는 다른 종류의 새롭게 배양된 세포를 감염시켜서 그 세포내에 증식되어 그 세포를 깨뜨림으로서 감염성이 있는 구제역바이러스로 생산된 단계, 혹은 다른 세포를 감염시키지 못하거나 감염시키더라도 더 이상 증식을 하지 못하는 단계 ;를 포함한다.
(7) Viral particles are combined with the RNA genome (base sequence of which (almost) matches the foot-and-mouth virus) and the expression of the foot-and-mouth particle protein expressed in the cell simultaneously in a manner that is difficult to find a difference with the general foot-and-mouth virus in the cell. The virus then infects another culture of the same or another type of newly cultured cell, and then proliferates within that cell, breaking the cell, producing the infectious foot-and-mouth virus, or preventing or infecting other cells. Even if the further proliferation step; includes.

이하, 본 발명을 도면을 참조하여 보다 구체적으로 설명한다.
Hereinafter, the present invention will be described in more detail with reference to the drawings.

도 1은 구제역 게놈이 벡터에 삽입된 형태를 나타낸 그림으로서 T7 promotor 아래에 바로 구제역 게놈서열이 시작되며 구제역 게놈 끝단의 poly (A) tail 뒤에는 제한효소 사이트(Nsi I)을 두어서 원형의 플라스미드를 단일의 제한효소 처리로 선형화 할 수 있도록 하였다. 따라서 이 제한효소는 이 원형의 플라스미드에서 오직 한 위치에만 존재하게 된다.
1 is a diagram showing the structure of the foot-and-mouth genome inserted into the vector. The foot-and-mouth genome sequence starts immediately under the T7 promotor, and the circular plasmid is placed with a restriction enzyme site (Nsi I) behind the poly (A) tail at the end of the foot-and-mouth genome. It was possible to linearize with a single restriction enzyme treatment. Thus, this restriction enzyme is only present at one position in this circular plasmid.

도 2는 도 1과 같이 구제역 전체 게놈이 cDNA 형태로 정상적으로 클로닝 되었을 때 나타나는 제한효소(Fsp I)의 소화된 형태를 나타낸다(약 6147, 2999, 1172, 759 크기의 네 개 단편이 확인 되어야 함). 이러한 패턴은 클론의 일부 유전자나 일부 서열을 수정(치환, 삭제 또는 삽입)하는 과정에서 그 조작이 가해진 산물에 대하여 그 조작의 전후에 있어서 클론의 기본적인 골격이 유지되었는지를 확인할 때 유용하다. 이렇게 그 기본골격이 유지된 클론에 대해 서열분석을 통해 의도한 돌연변이가 생성되었는지를 확인하게 되며, 연속되는 또는 추가적인 클로닝 과정중에 이러한 DNA 클론의 기본 골격이 깨진 군집(colony)는 선택에서 쉽게 배제되어 원하는 돌연변이의 선발을 보다 용이하게 한다.
Figure 2 shows the digested form of restriction enzyme (Fsp I) appears when the entire foot-and-mouth genome is normally cloned into cDNA form as shown in Figure 1 (four fragments of about 6147, 2999, 1172, 759 size should be identified) . This pattern is useful for confirming that the basic backbone of the clone was maintained before and after the manipulation of the product that had been manipulated in the process of modifying (substituting, deleting or inserting) some genes or sequences of the clone. The sequencing of the clones whose base skeleton was maintained confirms that the intended mutations were generated. During the subsequent or additional cloning process, colonies whose base skeleton of these DNA clones were broken were easily excluded from selection. Make selection of the desired mutations easier.

도 3은 최초의 구제역 cDNA 클론(P11)의 염기서열이 당초 공시된 바이러스의 서열과 비교할 때 CDR (coding region)에 상당한 비의도적 변화가 있음이 밝혀진 후에, P11의 CDR(coding region) 대부분을 교체하기 위해 선발되고 공시된 네 종류의 야외주 또는 세포 계대주 구제역바이러스와 이들 바이러스의 CDR으로 P11이 치환된 네 가지의 클론의 모식도를 나타낸다.
Figure 3 replaces most of the CDR regions of P11 after it has been found that the base sequence of the first foot-and-mouth disease cDNA clone (P11) has a significant unintentional change in the coding region (CDR) compared to the originally disclosed virus sequence. The schematics of the four types of field or cell passage foot-and-mouth disease virus selected and published for the purpose of the experiment and four clones in which P11 is substituted by CDRs of these viruses are shown.

도 4는 작성된 구제역 클론을 DNA 또는 RNA형태로 세포에 형질도입 시키는 과정과 형질도입후 게놈의 복제, 발현 및 감염성 획득여부에 따라 나타날 수 있는 결과에 대한 도해이다. 클론을 DNA형태로 도입시킬 경우에는 T7 RNA polymerase가 세포내에서 발현되도록 변형시킨 세포주(BHK T7-9, 일본 기후대학 Dr Ito 박사로부터 연구용으로 기증받음)를 활용하였다.
Figure 4 is a diagram showing the results of the process of transducing the prepared foot-and-mouth clones in the form of DNA or RNA and depending on whether the genome is cloned, expressed and infected after transduction. When the clone was introduced in the form of DNA, a cell line (BHK T7-9, donated for research by Dr. Ito, Ph.D., Japan Climatic University) was modified to express T7 RNA polymerase in cells.

도 5는 도 3과 같이 P11의 5' UTR(untranslated region) 대부분과 3‘ UTR을 그대로 유지하고 CDR을 교체하여 새롭게 작성한 네 종류의 클론(ASP, ASC, AS10 및 AS19)을 선형화하여 BHK T7-9세포에 transfection한 결과 AS10에서 구제역 바이러스 특이 구조단백질의 발현이 세 개 클론(ASP, ASC 및 AS10)에서 나타났고, AS10에서 가장 뚜렷하게 나타났다. 한편 상층액에서 바이러스의 RNA를 추출하여 실시간 RT-PCR로 정량분석을 실시한 결과 단백질의 발현량과 비례하는 수치로 RNA양이 관찰되었으며, AS19에서의 수치가 가장 낮았다. 따라서, AS10을 형질도입한 세포의 상층액에 감염성이 있는 구제역바이러스가 생성되어 있을 가능성이 가장 컸으나, 이들 상층액을 구제역감수성 세포에 접종시킨 결과 감염성은 확인되지 않았다. 비록 감염성은 확인되지 않았지만 클론별로 생산되는 RNA copy수의 차이 등을 볼 때 AS10, ASP 및 ASC 등은 세포내에서 replicon형태로 세포내에서 복제되고 구제역바이러스 단백질도 생산하고 있음을 확인할 수 있었다.
FIG. 5 shows the BHK T7 − by linearizing four newly cloned clones (ASP, ASC, AS10 and AS19) by maintaining most of the 5 'untranslated region (PTR) and 3' UTR of P11 as shown in FIG. Expression of foot-and-mouth virus-specific structural proteins in AS10 was observed in three clones (ASP, ASC and AS10), and most clearly in AS10. On the other hand, RNA was extracted from the supernatant and quantitatively analyzed by real-time RT-PCR. The amount of RNA was observed in proportion to the expression level of the protein, and the lowest in AS19. Therefore, infectious foot-and-mouth virus was most likely generated in the supernatant of cells transduced with AS10, but the infectivity was not confirmed as a result of inoculating these supernatants into the foot-and-mouth-sensitive cells. Although the infectivity was not confirmed, it was confirmed that AS10, ASP and ASC were cloned intracellularly in the form of replicon and produced foot-and-mouth disease virus protein by the difference in the number of RNA copies produced by the clones.

도 6은 가장 뚜렷한 replicon의 특징을 보이는, 공시된 구제역바이러스와 서열을 일치시킬 수록 감염력을 획득할 가능성이 높다는 기존의 문헌과 추론에 의거하여, 여전히 AS10의 5'UTR부위(대부분의 5'UTR은 클론 P11 유래임) 특정위치의 염기(IRES부위, 5‘ 끝단으로부터 665nt 번째, 기준 바이러스: )를 공시된 구제역바이러스 야외주에서 관찰되는 서열과 일치시키기 위해 Adenine에서 Guanine으로 치환 조작되거나 T7 RNA polymerase의 전사효율을 늘리고자 T7 promotor 끝단의 연속된 두 염기(GG)를 세 개로 연장한 클론의 세포내 형질도입 실험결과이다. 이러한 작업은 site-directed mutagenesis를 이용하였고 각각의 클론 조작결과를 IR 및 T7G3로 명명하였으며 두 종류의 돌연변이의 조합을 갖춘 조작결과는 T7G3IR로 명명하였다. 한편, 복제력을 상실한 클론을 작성하기 위하여 이미 보고된 바와 같이 3D polymerase의 일부를 제한효소, Sal I을 이용하여 삭제하였고 이 클론을 3Dmut으로 명명하고 세포내에서 복제가 불가능한 대조군으로 활용하였다. 새로이 작성된 클론들은 모두 전체염기서열을 분석하여 의도한 부위외에는 다른 돌연변이가 없음을 확인하였고, 선형화(線形)된 형태로 세포에 형질도입시킨 결과 도 6과 같이 대조 클론(3Dmut)과 비교할 때 다른 클론들에서는 뚜렷하게 증가된 복제현상을 확인할 수 있었다. 또한 이렇게 형질도입된 세포의 상층액에서 구제역바이러스 구조단백질이 발현되고 있음을 western blot으로 확인할 수 있었는데, 복제력을 상실시킨 AS10 유래의 대조 클론(3Dmut)도 세포에 전달시 구조단백질의 발현이 일부 확인되었다. 이는 바이러스 RNA 복제가 없어도 형질도입된 DNA로부터 전사된 RNA가 messenger RNA로 작동하여 일정량의 구제역바이러스 단백질을 발현시킬 수 있음을 보여주었다. 그러나 이들 모두에 있어서 2차 배양에서 세포의 변성을 일으키며 감염성을 보이는 현상은 확인되지 않았다. 한편, IR의 조작을 가할 경우 기존의 AS10 클론에 비교하여 복제나 발현능이 다소 상승되는 효과를 나타내었고 반대로 T7G3의 경우 오히려 AS10의 경우 보다 발현이나 복제능이 다소 떨어지는 현상을 보였으며 이는 T7G3IR의 경우도 마찬가지였다.
FIG. 6 is still the 5'UTR site of AS10 (most 5'UTRs), based on previous literature and inference that matching sequence with published foot-and-mouth virus, which is characterized by the most pronounced replicon, is more likely to obtain infectivity. Is derived from clone P11) A specific position (IRES site, 665nt from 5 'end, reference virus:) was engineered to substitute Guanine for Adenine or T7 RNA polymerase to match the sequence observed in the published foot and mouth virus virus The results of intracellular transduction experiments of clones that extended three consecutive two bases (GG) at the end of the T7 promotor to increase the transcriptional efficiency of. This work was performed using site-directed mutagenesis and each clone was named IR and T7G3, and the result of the combination of two mutations was named T7G3IR. On the other hand, in order to create clones that lost replication ability, as previously reported, a part of 3D polymerase was deleted using restriction enzyme, Sal I, and the clone was named as 3Dmut and used as a control that cannot be replicated in cells. All newly created clones were analyzed by the whole base sequence to confirm that there were no mutations other than the intended site. As a result of transducing the cells in a linearized form, the other clones were compared with the control clone (3Dmut) as shown in FIG. In the field, the replication phenomenon was clearly increased. In addition, Western blots showed that foot-and-mouth virus structural protein was expressed in the supernatant of the transduced cells. As a control clone (3Dmut) derived from AS10, which lost the replication ability, some of the structural protein expression was delivered to the cells. Confirmed. This demonstrated that RNA transcribed from transduced DNA could act as messenger RNA to express an amount of foot-and-mouth virus protein without viral RNA replication. However, in all of them, degeneration of cells in secondary culture and showing infectivity were not confirmed. On the other hand, when IR was manipulated, the replication or expression ability was slightly increased compared to the existing AS10 clone. On the contrary, T7G3 showed a somewhat lower expression or replication capacity than AS10, which is also the case with T7G3IR. It was the same.

도 7은 한편, 도 6에서 비교된 총 다섯 종의 구제역 cDNA클론을 in vitro transcription을 하여 RNA를 합성 후 서로 다른 세 종류의 세포에 도입시켜 발현 및 복제 양상을 비교해 본 것인데, BHK 세포에서는 구제역바이러스의 입자를 구성하는 구조단백질(VP1)의 발현을 웨스턴 블롯으로 확인할 수 있었다. 특히, T7 promotor 끝단에 한 개의 dGTP를 추가하여 총 세 개가 되게 하는 조작(T7G3)은 세포내에서의 복제 및 발현 효율을 떨어뜨니는 결과를 나타냈는데 이러한 결과는 도 6에서 선형화된 DNA형태로의 형질도입 실험결과와도 일치하는 것이었다. 이는 구제역바이러스 고유의 RNA 게놈의 5’ UTR 끝단에 원래 구제역바이러스 게놈에는 존재하지 않는 guadinine 3개가 불필요하게 따라 붙으면서 복제 및 발현에 오히려 저해되는 효과를 준 것으로 판단된다. 한편, 도 6과는 달리 RNA 게놈의 복제력을 고의적으로 상실시킨 AS10 유래의 대조 클론(3Dmut)에서는 RNA형태로의 세포내 형질 도입의 경우는 구조단백질 발현이 확인되지 않았다. 이러한 차이는 세포내에서 단백질 발현에 사용될 수 있는 messenger RNA의 상대적인 생성량(도입량)의 차이 혹은 messenger RNA의 세포내 이동경로나 분포의 차이에 의한 것으로 추정된다.
FIG. 7 is a comparison of expression and replication patterns by synthesizing RNA into three different types of cells after in vitro transcription of a total of five foot-and-mouth disease cDNA clones compared with those of FIG. 6, and in foot-and-mouth virus in BHK cells. Expression of the structural protein (VP1) constituting the particles of was confirmed by Western blot. In particular, the manipulation of adding one dGTP to the T7 promotor end to make a total of three (T7G3) resulted in a decrease in the efficiency of replication and expression in cells. The results were consistent with the transduction experiments. It is believed that this resulted in the inhibition of replication and expression by unnecessarily attaching three guadinines that were not present in the foot-and-mouth virus genome to the 5 'UTR end of the foot-and-mouth virus-specific RNA genome. On the other hand, unlike in FIG. 6, in the control clone (3Dmut) derived from AS10 which intentionally lost the replication power of the RNA genome, structural protein expression was not confirmed in the case of intracellular transduction into the RNA form. These differences may be due to differences in the relative production (introduction) of messenger RNA that can be used for protein expression in cells, or to differences in the intracellular migration pathway or distribution of messenger RNA.

도 8은 도 5에서 작성된 cDNA 클론중 증식성은 확인되었으나 감염성이 확인되지 않은 돼지유래 구제역바이러스 세포계대주 클론(ASP)의 추가 조작후 세포에 형질도입시킨 결과이다. 먼저 공시된 세포계대주 염기서열 결과에 비교할 때 s-fragment 및 poly (C) 부위에서 세포계대주의 염기서열과 불일치하는 P11 유래의 5'UTR 영역을 교정하기 위해 공시된 세포주 유래 구제역바이러스의 해당부위를 증폭하여 클로닝한 뒤 ASP의 해당부위를 교체하는 데 성공하였고 이를 ASPs12L 클론으로 명명하였고, poly(C) tract의 cytidine의 개수는 여기서 12개였다. 여기서 poly(C) segment의 cytidine의 개수를 늘려서 감염력의 획득에 보다 유리한 조건을 충족시킨 바, 이 클론을 ASPs24L로 명명하였다. 이 클론의 poly(C) tract의 정확한 길이는 capillary sequencing 기술로는 단정하기 어려우며 최소 24개에서 최대 36개로 추정된다. 이 클론(ASPs24L)의 경우 공시된 구제역바이러스 세포계대주의 서열 대비 synonymous mutation은 6개가 관찰되었고 non-synonymous mutation은 1개가 관찰되었다. 여기서 아미노산 차이는 VP1의 34번째 아미노산에 해당되었다. ASPs24L클론을 선형화된 DNA의 형태로 BHK T7-9 세포에 형질도입한 이후 세포변성효과가 7일경부터 관찰되었고 이 상층액을 IBRS-2세포에 감염시킨 결과 감염력이 확인되었다.
FIG. 8 is a result of transducing cells after further manipulation of swine-based foot-and-mouth virus virus subpopulation clones (ASPs) of which the proliferability was confirmed but not confirmed in the cDNA clones prepared in FIG. 5. Compared to the cell lineage sequence results disclosed above, the corresponding site of the cell line-derived foot-and-mouth virus that was disclosed to correct the P11-derived 5'UTR region at the s-fragment and poly (C) sites inconsistent with the cell lineage sequence After amplification and cloning, we replaced the corresponding site of ASP and named it ASPs12L clone, and the number of cytidine of poly (C) tract was 12 here. Here, the clone was named ASPs24L because it increased the number of cytidine in the poly (C) segment to satisfy the more favorable condition for obtaining infectivity. The exact length of the poly (C) tract of this clone is difficult to determine with capillary sequencing techniques and is estimated to be from 24 to 36. In this clone (ASPs24L), six synonymous mutations and one non-synonymous mutation were observed relative to the published foot-and-mouth virus cell line sequence. The amino acid difference corresponds to the 34th amino acid of VP1. After transducing the ASPs24L clone into BHK T7-9 cells in the form of linearized DNA, cytopathic effects were observed from day 7 and infecting the supernatant with IBRS-2 cells confirmed the infectivity.

도 9는 이렇게 감염된 세포의 상층액을 수거하여 VP1부위의 염기서열을 조사 비교한 결과로서 ASPs24L클론이 공시된 세포계대주 대비 아미노산서열에서 유일하게 상이했던 VP1의 non-synonymous mutation site가 이 클론을 통하여 IBRS-2의 상층액에서 획득한 바이러스에서는 공시된 세포계대주와 일치하였는데, 이는 무작위적 생성 또는 무작위적 발생과 결부된 선택의 결과로 해석될 수 있을 것이다. 이렇게 최초 IBRS-2 세포주 상층액은 약 3x104 PFU/ml의 역가를 나타내었다. 한편, poly(C) tract의 연장없이 site-directed mutagensis를 통하여 VP1의 34번째 아미노산 서열만을 분리주의 아미노산 서열(F)과 일치시킨 클론 (ASPs11F)도 감염성이 있음을 확인하였다.
9 is a non-synonymous mutation site of VP1 which was uniquely different from the amino acid sequence compared to the cell line where the ASPs24L clone was disclosed as a result of collecting the supernatant of the infected cells and examining the nucleotide sequence of the VP1 region. Virus obtained from the supernatant of IBRS-2 was consistent with the published cell lines, which could be interpreted as the result of selection associated with random generation or random development. The initial IBRS-2 cell line supernatant thus showed a titer of about 3 × 10 4 PFU / ml. On the other hand, the site-directed mutagensis without poly (C) tract extension confirmed that the clone (ASPs11F), which only matched the amino acid sequence (F) of the 34th amino acid sequence of VP1, was also infectious.

이하, 본 발명을 실시예 및 시험예를 통하여 보다 구체적으로 설명하지만, 본 발명이 이에 의해 한정되는 것은 아니고, 당업자에서 통상적으로 주지된 변형, 치환 및 삽입 등을 수행할 수 있으며, 이에 대한 것도 본원 발명의 범위에 포함된다.
Hereinafter, the present invention will be described in more detail with reference to examples and test examples, but the present invention is not limited thereto, and modifications, substitutions, and insertions that are commonly known by those skilled in the art can be performed. It is included in the scope of the invention.

[실시예 1] P11을 기본 골격으로 하는 replicon 수준 구제역 cDNA 클론의 작성
Example 1 Preparation of replicon level foot-and-mouth disease cDNA clones based on P11

2002년 국내분리 O형 구제역바이러스를 감염시킨 IBRS-2 세포 상층액을 RNeasy extraction 키트(Qiagen)를 사용하여 바이러스 RNA를 추출하였다. 상보적인 DNA는 random hexamer 또는 oligo dT를 프라이머로 하여 Superscript III (Invitrogen)를 사용하여 제작하였고, cDNA를 주형으로 Expand Long template PCR system (Roche) 또는 LA Taq(Takara)를 이용하여 구제역 게놈 전체영역을 4개로 나누어 증폭한 뒤 각각을 클로닝하고 4개의 클론을 순차적으로 연결하여 최종적인 클론(P11)을 얻었다. P11 클론은 정확도(fidelity)가 높지 않은 Taq polymerase의 사용과 지나치게 많은 단계의 클로닝과정 때문에 아미노산 수준에서 의도하지 않았던 돌연변이가 나타났고, 특히 AY312589.1의 여기서열과 비교할 때 10개 이상이 달랐고, 5‘UTR의 변화도 생겨났다.
Viral RNA was extracted from IBRS-2 cell supernatants infected with domestically isolated O-type foot-and-mouth virus in 2002 using the RNeasy extraction kit (Qiagen). Complementary DNA was prepared using Superscript III (Invitrogen) using a primer of random hexamer or oligo dT, and the entire foot-and-mouth genome region was constructed using Expand Long template PCR system (Roche) or LA Taq (Takara) as a template for cDNA. After amplification into four, each cloned and four clones were sequentially connected to obtain a final clone (P11). The P11 clone had unintended mutations at the amino acid level due to the use of non-fidelity Taq polymerase and too many steps of cloning, especially over 10 different in comparison to the AY312589.1 here column. UTRs have also changed.

따라서, 우리는 P11 클론을 교정하기 위해서 에러율이 아주 적은 Phusion High Fidelity DNA polymerase(Finnzyme)를 이용하여 공시된 네 종류의 바이러스의 특정부위를 폭하였다. 증폭에 사용된 순방향 프라이머(xba-pst-F)의 서열은 5' CCTAGTTTTGCCCGTTTTCA 3'이고 역방향 프라이머(xba-pst-R)의 서열은 5' GCCACTAGT TTATTGTTTCTCGACGCCAAT 3'이다. 증폭된 산물을 제한효소 Xba I와 Pst I으로 소화시키면 약 6730bp에 달하는 구제역바이러스 게놈의 약 82%에 해당하는 insert를 얻을 수 있고 벡터는 P11클론을 역시 Xba I과 Pst I로 소화시켜 얻었다.. 이들 분해 산물을 agarose gel상에서 분리 정제한 후 Long ligation kit(TAKARA, Japan)을 이용하여 ligation 시키고 에탄올 precipitation으로 다시 정제한 후, ElectroMAX DH10B competent cell(Invitrogen)에 electroporation 법으로 도입한 후 amplicilin이 함유된 세균배지에 도말하여 하루 이상 37℃ 배양기에 배양하여 최종적으로 형질전환된 군집(colony)를 찾았다. Colony에 대한 PCR을 통해서 일단 구제역바이러스 게놈이 삽입된 것으로 추정되는 DNA클론을 선발하였다. 그리고 종류(AS10, AS19, ASP, ASC)별로 최종 선발된 군집(colony)을 다시금 24시간 이상 200ml 이상의 액체배지에 배양하여 이 대장균들에서 midiprep을 실시하여 약 100ug 이상의 DNA 클론을 얻었다. 이렇게 획득된 DNA 클론은 FSP I 제한효소로 처리하여 DNA클론의 크기와 방향성이 올바른 지 확인된 이후(도 2), DNA 클론 전체 부위에 대한 서열분석을 실시하여 의도한 돌연변이가 제대로 유도되었는 지를 확인하였다 (AS10, AS19, ASP, ASC 각 서열정보).
Therefore, we used the Phusion High Fidelity DNA polymerase (Finnzyme) with a very low error rate to correct the P11 clones, thus defining four specific viruses. The sequence of the forward primer (xba-pst-F) used for amplification is 5 'CCTAGTTTTGCCCGTTTTCA 3' and the sequence of the reverse primer (xba-pst-R) is 5 'GCCACTAGT TTATTGTTTCTCGACGCCAAT 3'. Digestion of the amplified product with restriction enzymes Xba I and Pst I yielded an insert corresponding to about 82% of the AML virus genome of about 6730 bp, and the vector was digested with P11 clones by Xba I and Pst I. These products were separated and purified on agarose gel, ligation using long ligation kit (TAKARA, Japan), purified by ethanol precipitation, introduced into electroMAX DH10B competent cell (Invitrogen) by electroporation, and then containing amplicilin. The plated on bacterial medium was cultured in an incubator at 37 ° C. for at least one day to find a finally transformed colony. PCR for colony was used to select DNA clones once believed to contain the foot and mouth virus genome. The colonies finally selected for each type (AS10, AS19, ASP, ASC) were again incubated in a liquid medium of 200 ml or more for more than 24 hours and subjected to midiprep of the E. coli to obtain a DNA clone of about 100 ug or more. The DNA clones thus obtained were treated with FSP I restriction enzymes to confirm that the size and orientation of the DNA clones were correct (FIG. 2), followed by sequencing of the entire region of the DNA clones to confirm that the intended mutations were properly induced. (SEQ ID information of AS10, AS19, ASP, ASC).

각각의 DNA 클론중에서 세포에 전달시 복제 및 구제역단백질 발현이 확인되는 AS10클론을 site-directed mutageneiss kit (GenTailer, Invitrogen)을 이용하여 T7 promotor 뒤에 G의 길이를 2개에서 3개로 연장하고 이에 대하여 다시 IRES의 특정부위의 서열을 공시된 바이러스의 서열과 일치시켜 교체하였다.
In each of the DNA clones, the AS10 clone, whose replication and foot-and-mouth protein expression was confirmed upon delivery to the cell, was extended to two to three G lengths after the T7 promotor using the site-directed mutageneiss kit (GenTailer, Invitrogen). The sequence of the specific site of the IRES was replaced by matching the sequence of the published virus.

[실시예 2] ASP를 기본 골격으로 하는 감염성 구제역 cDNA 클론의 작성
Example 2 Preparation of Infectious Foot-and-mouth disease cDNA Clones Based on ASP

ASP 클론의 S fragment를 site-directed mutation을 시킨 클론(ASPs12L)의 poly (C) region을 최대 36개까지 연장시키기 위해 특별히 제작된 primers 를 사용하였다. 이는 poly(C)의 길이가 길어질수록 바이러스의 병원성과 상관성이 있을 수 있다는 발표와 관련이 있다. 이렇게 얻어진 클론의 poly(C) region이 얼마나 연장되었는가는 capillary sequencing 방법을 통해서는 정확히 그 길이를 가늠할 수는 없었지만 기존에 12개 이었던 poly (C)의 길이가 최소 24개 이상으로 연장된 것은 확실히 판단할 수 있었으며 최대 36개 일 것으로 추정된다. 이 클론을 ASPs24L로 명명하였다. 여기서 L은 VP1의 34번째 아미노산으로서 공시된 구제역바이러스(돼지유래세포계대주)의 아미노산서열(F)의 불일치가 일어나는 유일한 곳으로서 아직 F로 치환되지 않은 상태를 나타낸다. 이 ASPs24L은 DNA 및 RNA 형태로 세포에 형질도입시 모두 감염력이 있는 바이러스가 생산되는 것이 확인되었다.
Specially designed primers were used to extend up to 36 poly (C) regions of the clones (ASPs12L) that site-directed mutation of the S fragment of the ASP clone. This is related to the announcement that longer poly (C) may correlate with the pathogenicity of the virus. The length of the poly (C) region of the clone thus obtained could not be accurately determined by the capillary sequencing method, but it was certainly judged that the length of the previously 12 poly (C) was extended to at least 24. It is possible to do so and is estimated to be up to 36. This clone was named ASPs24L. Where L is the only place where the discrepancy of the amino acid sequence (F) of foot-and-mouth virus (swine-derived cell lineage) published as the 34th amino acid of VP1 occurs and is not yet substituted with F. The ASPs24L was confirmed to produce infectious viruses when transduced into cells in the form of DNA and RNA.

[실시예 3] ASP클론에서 감염성 바이러스를 얻기 위한 충분조건인 VP1의 34번째 아미노산 서열(F) 및 감염력 획득에 있어서 poly(C)의 역할
Example 3 34th amino acid sequence (V) of VP1 which is sufficient condition for obtaining infectious virus in ASP clone and role of poly (C) in obtaining infectivity

한편, 감염력을 획득한 바이러스는 VP1의 34번째 아미노산이 L에서 F로 치환되어 있었다. 따라서, ASPs35L 클론의 VP1 34번째 아미노산인 L을 F로 site-directed mutagenesis를 통해서 조작한 결과 VP1 34번째 아미노산 L이 F로 변환되었으나 poly(C)의 길이가 11개로 줄어들어 있었다. ASPs11F로 명명된 이 클론도 세포실험결과 감염성을 나타내었다. 따라서 poly (C)의 길이가 감염력의 획득에 중요한 역할을 하는 것이 인정되지만, poly(C)의 길이가 11개 일때도 감염력을 나타내는 것을 볼 때 poly(C) tract의 역할은 바이러스의 RNA genome의 복제과정 중 바이러스의 생존에 필요한 돌연변이의 발생 및 선발을 촉발하는 기능에 일조하는 것으로 추정된다.
On the other hand, in the virus which obtained infectivity, the 34th amino acid of VP1 was substituted from L to F. Therefore, the 34th amino acid L of the VP1 35L clone was manipulated by site-directed mutagenesis to F, but the 34th amino acid L of VP1 was converted to F, but the length of poly (C) was reduced to 11. This clone, named ASPs11F, also showed infectivity in cell experiments. Therefore, although the length of poly (C) plays an important role in the acquisition of infectivity, the role of poly (C) tract may be due to the viral genome of 11 viruses. It is believed to contribute to the ability to trigger the development and selection of mutations necessary for the survival of the virus during replication.

[실시예 4] 구제역바이러스 게놈 cDNA 플라스미드와 이 플라스미드가 복제되는 숙주대장균의 증식과의 상관성 확인
Example 4 Confirmation of correlation between foot-and-mouth virus genome cDNA plasmid and host E. coli to which this plasmid is replicated

이러한 현상은 바이러스와 진핵세포와의 상호작용의 결과가 아닌 바이러스 게놈이 담긴 플라스미드와 이 플라스미드가 복제되는 곳인 대장균(competent cell)과의 상호작용 결과에 대한 것이다. 이는 바이러스 게놈이 대장균내에서서 circular한 DNA 이중나선 구조로 존재하면서 일정 정도 transcription이 일어나 mRNA형태로 전사되고 이중 일부는 또 단백질로 번역되면서 세균의 생장에 부정적인 역할을 할 수 있다는 추정을 하게된다. 이는 DNA 클론에 돌연변이가 성공적으로 가해져 공시된 바이러스의 게놈 서열과 보다 완벽하게 일치될수록 이렇게 성공된 플라스미드가 담긴 형질전환된 대장균의 colony는 상대적으로 그 크기가 작거나 생장이 느려지는 경우가 많기 때문이다. 따라서, poly(C) tract가 연장된 colony를 대량배양하여 상당한 양의 플라스미드를 정제해 내기 위해서는 변형된 방법으로 이들을 배양할 필요가 있다. 이렇게 poly(C)가 연장된 플라스미드가 확인된 후 대량으로 이들을 정제회수하기 위해서는, 이 프라스미드를 다시 대장균에 transformation하여 세균배지에 도말하여 선발하는 과정을 거치지 않고 1시간정도 transformation된 대장균을 약 1ml 정도의 액체배지에서 진탕배양한 다음 바로 이 배양액을 200ml 이상의 달하는 액체배지에 부어서 1-2일간 낮은 rpm에서 생장시키는 것이다. 이러한 방법으로 보다 성공적으로 형질전환된 세균으로 부터 불안정하고 생장을 더디게 만드는 구제역바이러스 전체 게놈서열이 정확한 순서와 구성으로 담긴 플라스미드를 대량으로 얻어 실험에 활용할 수 있었다.
This is not a result of the interaction between the virus and the eukaryotic cell, but the result of the interaction between the plasmid containing the viral genome and the competent cell, where it is replicated. It is assumed that the viral genome exists as a circular DNA double-helix structure in Escherichia coli, which causes transcription to some extent and is transcribed into mRNA, some of which may also be translated into proteins, negatively affecting the growth of bacteria. This is because colony of transformed Escherichia coli containing such a successful plasmid is relatively small in size or slowed down in growth as the mutation of the DNA clone is successfully matched with the published genome sequence. . Therefore, in order to purify a considerable amount of plasmids by mass culturing poly (C) tract extended colonies, it is necessary to culture them by a modified method. In order to purify the poly (C) -extended plasmids and to recover them in large quantities, about 1 ml of the transformed E. coli transformed for about 1 hour without undergoing the process of transforming the plasmids into E. coli and smearing them into a bacterial medium. After shaking the culture in the liquid medium of the degree, this culture is poured into 200 ml or more of liquid medium and grown at low rpm for 1-2 days. In this way, the entire genome sequence of foot-and-mouth virus, which is unstable and slow to grow from more successfully transformed bacteria, was obtained in large quantities and used in experiments.

이상 본 발명의 구체적 실시형태와 관련하여 본 발명을 설명하였으나 이는 예시에 불과하며 본 발명은 이에 제한되지 않는다. 당업자는 본 발명의 범위를 벗어나지 않고 설명된 실시형태를 변경 또는 변형할 수 있으며, 이러한 변경 또는 변형도 본 발명의 범위에 속한다. 또한, 본 명세서에서 설명한 각 구성요소의 물질은 당업자가 공지된 다양한 물질로부터 용이하게 선택하여 대체할 수 있다. 또한 당업자는 본 명세서에서 설명된 구성요소 중 일부를 성능의 열화 없이 생략하거나 성능을 개선하기 위해 구성요소를 추가할 수 있다. 뿐만 아니라, 당업자는 공정 환경이나 장비에 따라 본 명세서에서 설명한 방법 단계의 순서를 변경할 수도 있다. 따라서 본 발명의 범위는 설명된 실시형태가 아니라 특허청구범위 및 그 균등물에 의해 결정되어야 한다.
Although the present invention has been described in connection with the specific embodiments of the present invention, it is to be understood that the present invention is not limited thereto. 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 as defined by the appended claims. In addition, the materials of each component described herein can be readily selected and substituted for various materials known to those skilled in the art. Those skilled in the art will also appreciate that some of the components described herein can be omitted without degrading performance or adding components to improve performance. In addition, those skilled in the art may change the order of the method steps described herein depending on the process environment or equipment. Therefore, the scope of the present invention should be determined by the appended claims and equivalents thereof, not by the embodiments described.

<110> Animal, Plant and Fisheries Quarantine and Inspection Agency <120> Infectious cDNA clones of Foot-and-mouth disease virus of type O and the complete sequences of the clones <130> PH11-8663 <160> 6 <170> KopatentIn 2.0 <210> 1 <211> 11093 <212> DNA <213> cDNA clone <400> 1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtgctagta acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaagct tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gagcacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta 1800 aaacactgtt cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gttcaacgga ggatgcttgt tggtggccat ggtgccagaa ctttgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacgtgtt gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg ttcacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aatccacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgag ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgt 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tgcaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggctcgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg cgagagcccc gtgaccaatc tgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ctgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagtccaa ccctggaccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccat caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa gactatcagg accggtctcg acgaggccaa accctggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg 5100 ccattctcaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac agacctggtg cctggcatcc 5220 ttgaaaagca acaggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag aaccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt tgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaagg tcatcatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aagacatgtt caagccccaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccgggt cgagctccac gagaaggtgt 6000 cgaaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgaattctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacccc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acactgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaga gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttgg cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 2 <211> 11093 <212> DNA <213> ASP clone <400> 2 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtactagta acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaagct tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gagcacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta 1800 aaacactgtt cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gttcaacgga ggatgcttgt tggtggccat ggtgccagaa cattgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacatgtt gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg ttcacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aattcacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgaa ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgc 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tgcaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggcttgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg caagagcccc gtgaccaatc tgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ctgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagtccaa ccctgggccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccat caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa ggctatcagg accggtctcg acgaggccaa accatggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg 5100 ccattctcaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac agacctagtg cctggcatcc 5220 ttgaaaagca gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag aaccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt cgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaagg tcatcatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aagacatgtt caagcctcaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccgggt cgagctccac gagaaggtgt 6000 cgaaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgaattctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acactgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 3 <211> 11093 <212> DNA <213> ASC clone <400> 3 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtgctagta acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaaact tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gaccacaacc gactgtttca tcgctttgtt gtacgcctcc agagagatta 1800 aaacactgtt cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgggggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gctcaacgga ggatgcttgt tggtggccat ggtgccagaa ctttgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcga 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacatgtt gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg tttacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aattcacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgag ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgt 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tacaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacaagg gggacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggcttgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg cgagagcccc gtgaccaatc cgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ttgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgagaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagcccaa ccctgggccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccgt caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa ggccatcagg accggtctcg acgaggccaa accctggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcggc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg cgacatcaat gacatattcg 5100 ccattcttaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac agacctggtg cctggcatcc 5220 ttgaaaagca gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag caccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt tgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaggg tcatcatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aaaacatgtt caagccccaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccaggt cgagctccac gagaaggtgt 6000 cggaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgacttctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatggttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acattgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 4 <211> 11093 <212> DNA <213> AS19 clone <400> 4 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtgctagtg acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaaact tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gaccacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta 1800 aaacaccgtt cttatcacga gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gttcaacgga ggatgcttgt tggtggccat ggtgccagaa ctttgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacatgtt gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg tttacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aattcacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgct tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgag ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgt 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tacaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggcttgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg cgagagcccc gtgaccaatc cgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ttgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagcccaa ccctgggccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccgt caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa ggccatcagg accggtctcg acgaggccaa accctggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg 5100 ccattcttaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac agacctggtg cctggcatcc 5220 ttgaaaagca gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagcggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag caccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt tgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaagg tcataatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aagacatgtt caagccccaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccaggt cgagctccac gagaaggtgt 6000 cggaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgacttctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acattgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagggccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cctccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 5 <211> 11093 <212> DNA <213> AS10 clone <400> 5 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtgctagta acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaagct tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gagcacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta 1800 aaacactgtt cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gttcaacgga ggatgcttgt tggtggccat ggtgccagaa ctttgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacatgtt gccggggcgg ttcaccaacc tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg ttcacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aattcacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgag ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgt 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tgcaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggcttgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg cgagagcccc gtgaccaatc tgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ctgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagtccaa ccctgggccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccat caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa ggctatcagg accggtctcg acgaggccaa accctggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg 5100 ccattctcaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac ggacctggtg cctggcatcc 5220 ttgaaaagca gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag aaccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt tgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaagg tcatcatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aagacatgtt caagccccaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccgggc cgagctccac gagaaggtgt 6000 cgaaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgaattctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acactgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagcgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 6 <211> 11083 <212> DNA <213> ASPs12L clone <400> 6 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgggc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gggccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccacttg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctacttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccccc 1020 ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc ttgcgactgt 1080 cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg gcgttaattg 1140 gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc ctagttttgc 1200 ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg aggaggactt 1260 gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac ttgaaactcc 1320 gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact ccacgctcgg 1380 tccactggcg agtactagta acagcactgc tgcttcgtag cggagcatgg tggccgcggg 1440 aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac ggacccacca 1500 tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg acactgatac 1560 tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga ggtaacacgc 1620 gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt ttaaaaagct 1680 tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact atttttaaat 1740 gagcacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta aaacactgtt 1800 cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga agaaaacatt 1860 ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc agttgtttag 1920 gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc tcacgctcga 1980 tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg gaccacccgc 2040 tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg cttcgcgacc 2100 cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc acgctggcat 2160 cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg ggtggtacgc 2220 gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc tggtgttcgt 2280 cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga aacgactcag 2340 aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta acactggaag 2400 cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac aacttggtga 2460 caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca cccacacaac 2520 caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta gcggtctttt 2580 cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg accgcatcct 2640 cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag tcacttacgg 2700 gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg agaccagggt 2760 tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca gtgacccgtt 2820 cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg gcagcctgac 2880 cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg tgggaaatca 2940 gttcaacgga ggatgcttgt tggtggccat ggtgccagaa cattgctcta ttgacaagag 3000 agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga cgaacatgac 3060 ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca aagtacacaa 3120 accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg aaggtgcccc 3180 acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg gtgagctccc 3240 ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc tggtgaccac 3300 tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc gcaacatgtt 3360 gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt tcctgcactt 3420 tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac tcgcccagtt 3480 tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc tcgcccagta 3540 ctacacacag tacagcggca ccatcaacct gcacttcatg ttcacaggac ccactgacgc 3600 gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca aaacacccga 3660 ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa aattcacatt 3720 ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg ccgcggagac 3780 cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg ctgacggcga 3840 cgcactggtc gttctagcta gcgccggtaa ggactttgaa ctgcgtctgc cagttgacgc 3900 tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca ccgttgagaa 3960 ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgc tcatactaga 4020 cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga tgcaaatccc 4080 tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg cagatctgga 4140 agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc ccgaggcagc 4200 gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc ggcttgcact 4260 gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact gcaagtatgg 4320 caagagcccc gtgaccaatc tgagaggtga cctgcaagtg ttgacccaga aggcggcaag 4380 aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga ctgaactgct 4440 ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta ttcacccgag 4500 cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga actttgacct 4560 gctcaagttg gcaggagacg tcgagtccaa ccctgggccc ttcttcttct ctgacgtcag 4620 gtcaaatttt tccaagttgg ttgaaaccat caaccagatg caggaggaca tgtcaacaaa 4680 acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca ctggagtgaa 4740 ggctatcagg accggtctcg acgaggccaa accatggtac aagctcatca agctcttgag 4800 ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc ttgtggccat 4860 catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga agaagatctc 4920 cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg gagccccgat 4980 tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac ccgaagacct 5040 tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg ccattctcaa 5100 gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca aggcatggat 5160 cgcctcagaa gaaaagtttg tcaccatgac agacctagtg cctggcatcc ttgaaaagca 5220 gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg acaacgcgcg 5280 ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag tggttgcccc 5340 agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca agtcaggcca 5400 gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact tcactggcag 5460 aaccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt acaaccagca 5520 gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact tcaagtactt 5580 cgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg aggacaaagg 5640 caagcctttt aacagcaagg tcatcatcgc caccaccaac ctgtactcgg gcttcacccc 5700 gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca ttgacgtgag 5760 cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc ttgaagacac 5820 ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg gcatggccgt 5880 tgaaatgaag agaatgcaac aagacatgtt caagcctcaa ccgcccctcc agaacgtcta 5940 ccagcttgtt caggaggtga ttgaccgggt cgagctccac gagaaggtgt cgaaccaccc 6000 gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc tcattgagaa 6060 aggccagcac gaagcagcaa ttgaattctt tgaggggatg gtgtgtgact ccatcaagga 6120 ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta agcgcctgaa 6180 ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag tgatcatgat 6240 ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt acactgagaa 6300 ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc ctctggaaac 6360 cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca agacgggtga 6420 tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg aaggacccta 6480 caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc cgcagcagga 6540 ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag cgaaagcccc 6600 ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt tgaaggtgaa 6660 agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc aaaagatggt 6720 catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg ccatctgctg 6780 cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg cagagaaata 6840 tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag tgtttgagtt 6900 tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg tgcttcaccg 6960 tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa tgaagaaagg 7020 cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct tctctggtga 7080 ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc ccggtctctt 7140 cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg caaaggacgg 7200 agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg gatactgctc 7260 atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg aaccacacca 7320 cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca tgcgcaaaac 7380 caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc ctgccgcctt 7440 gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca tcttctccaa 7500 acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc gctgtgctgc 7560 tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac tgagcattta 7620 cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg cgcctggtct 7680 cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga acggcactgt 7740 cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca agtttgtatg 7800 ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg gtaagactcg 7860 cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga ttggcagatt 7920 ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg gttgtaatcc 7980 tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg tgtgggatgt 8040 ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca tgtttgagga 8100 ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga aaactctcgt 8160 gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga tgccgtctgg 8220 ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc tctacgcctt 8280 gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct acggagacga 8340 catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc acttcaaatc 8400 ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc ttggtcactc 8460 cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa ctgggtttta 8520 caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac gtcgtgggac 8580 catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac ctgacgagta 8640 ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca gatcacttta 8700 cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc acaattggca 8760 gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac tttttcccgc 8820 ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc atgtattcta 8880 tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc atggtcatag 8940 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 9000 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 9060 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 9120 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 9180 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 9240 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 9300 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 9360 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 9420 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 9480 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 9540 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 9600 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 9660 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 9720 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 9780 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 9840 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 9900 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 9960 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 10020 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 10080 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 10140 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 10200 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 10260 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 10320 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 10380 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 10440 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 10500 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 10560 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 10620 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 10680 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 10740 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 10800 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 10860 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 10920 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 10980 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 11040 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11083 <110> Animal, Plant and Fisheries Quarantine and Inspection Agency <120> Infectious cDNA clones of Foot-and-mouth disease virus of type O          and the complete sequences of the clones <130> PH11-8663 <160> 6 <170> Kopatentin 2.0 <210> 1 <211> 11093 <212> DNA <213> cDNA clone <400> 1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtgctagta acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaagct tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gagcacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta 1800 aaacactgtt cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gttcaacgga ggatgcttgt tggtggccat ggtgccagaa ctttgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacgtgtt gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg ttcacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aatccacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgag ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgt 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tgcaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggctcgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg cgagagcccc gtgaccaatc tgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ctgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagtccaa ccctggaccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccat caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa gactatcagg accggtctcg acgaggccaa accctggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg 5100 ccattctcaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac agacctggtg cctggcatcc 5220 ttgaaaagca acaggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag aaccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt tgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaagg tcatcatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aagacatgtt caagccccaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccgggt cgagctccac gagaaggtgt 6000 cgaaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgaattctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacccc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acactgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaga gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttgg cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 2 <211> 11093 <212> DNA <213> ASP clone <400> 2 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtactagta acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaagct tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gagcacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta 1800 aaacactgtt cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gttcaacgga ggatgcttgt tggtggccat ggtgccagaa cattgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacatgtt gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg ttcacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aattcacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgaa ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgc 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tgcaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggcttgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg caagagcccc gtgaccaatc tgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ctgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagtccaa ccctgggccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccat caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa ggctatcagg accggtctcg acgaggccaa accatggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg 5100 ccattctcaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac agacctagtg cctggcatcc 5220 ttgaaaagca gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag aaccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt cgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaagg tcatcatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aagacatgtt caagcctcaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccgggt cgagctccac gagaaggtgt 6000 cgaaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgaattctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acactgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 3 <211> 11093 <212> DNA <213> ASC clone <400> 3 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtgctagta acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaaact tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gaccacaacc gactgtttca tcgctttgtt gtacgcctcc agagagatta 1800 aaacactgtt cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgggggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gctcaacgga ggatgcttgt tggtggccat ggtgccagaa ctttgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcga 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacatgtt gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg tttacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aattcacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgag ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgt 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tacaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacaagg gggacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggcttgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg cgagagcccc gtgaccaatc cgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ttgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgagaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagcccaa ccctgggccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccgt caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa ggccatcagg accggtctcg acgaggccaa accctggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcggc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg cgacatcaat gacatattcg 5100 ccattcttaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac agacctggtg cctggcatcc 5220 ttgaaaagca gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag caccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt tgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaggg tcatcatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aaaacatgtt caagccccaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccaggt cgagctccac gagaaggtgt 6000 cggaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgacttctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatggttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acattgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 4 <211> 11093 <212> DNA <213> AS19 clone <400> 4 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtgctagtg acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaaact tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gaccacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta 1800 aaacaccgtt cttatcacga gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gttcaacgga ggatgcttgt tggtggccat ggtgccagaa ctttgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacatgtt gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg tttacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aattcacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgct tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgag ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgt 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tacaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggcttgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg cgagagcccc gtgaccaatc cgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ttgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagcccaa ccctgggccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccgt caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa ggccatcagg accggtctcg acgaggccaa accctggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg 5100 ccattcttaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac agacctggtg cctggcatcc 5220 ttgaaaagca gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagcggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag caccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt tgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaagg tcataatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aagacatgtt caagccccaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccaggt cgagctccac gagaaggtgt 6000 cggaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgacttctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acattgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagggccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cctccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 5 <211> 11093 <212> DNA <213> AS10 clone <400> 5 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgagc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gagccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccactcg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctatttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccctc 1020 cccccccccc ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc 1080 ttgcgactgt cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg 1140 gcgttaattg gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc 1200 ctagttttgc ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg 1260 aggaggactt gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac 1320 ttgaaactcc gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact 1380 ccacgctcgg tccactggcg agtgctagta acagcactgc tgcttcgtag cggagcatgg 1440 tggccgcggg aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac 1500 ggacccacca tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg 1560 acactgatac tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga 1620 ggtaacacgc gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt 1680 ttaaaaagct tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact 1740 atttttaaat gagcacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta 1800 aaacactgtt cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga 1860 agaaaacatt ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc 1920 agttgtttag gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc 1980 tcacgctcga tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg 2040 gaccacccgc tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg 2100 cttcgcgacc cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc 2160 acgctggcat cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg 2220 ggtggtacgc gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc 2280 tggtgttcgt cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga 2340 aacgactcag aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta 2400 acactggaag cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac 2460 aacttggtga caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca 2520 cccacacaac caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta 2580 gcggtctttt cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg 2640 accgcatcct cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag 2700 tcacttacgg gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg 2760 agaccagggt tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca 2820 gtgacccgtt cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg 2880 gcagcctgac cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg 2940 tgggaaatca gttcaacgga ggatgcttgt tggtggccat ggtgccagaa ctttgctcta 3000 ttgacaagag agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga 3060 cgaacatgac ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca 3120 aagtacacaa accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg 3180 aaggtgcccc acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg 3240 gtgagctccc ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc 3300 tggtgaccac tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc 3360 gcaacatgtt gccggggcgg ttcaccaacc tccttgatgt ggctgaggcg tgccctacgt 3420 tcctgcactt tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac 3480 tcgcccagtt tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc 3540 tcgcccagta ctacacacag tacagcggca ccatcaacct gcacttcatg ttcacaggac 3600 ccactgacgc gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca 3660 aaacacccga ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa 3720 aattcacatt ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg 3780 ccgcggagac cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg 3840 ctgacggcga cgcactggtc gttctagcta gcgccggtaa ggactttgag ctgcgtctgc 3900 cagttgacgc tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca 3960 ccgttgagaa ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgt 4020 tcatactaga cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga 4080 tgcaaatccc tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg 4140 cagatctgga agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc 4200 ccgaggcagc gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc 4260 ggcttgcact gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact 4320 gcaagtatgg cgagagcccc gtgaccaatc tgagaggtga cctgcaagtg ttgacccaga 4380 aggcggcaag aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga 4440 ctgaactgct ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta 4500 ttcacccgag cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga 4560 actttgacct gctcaagttg gcaggagacg tcgagtccaa ccctgggccc ttcttcttct 4620 ctgacgtcag gtcaaatttt tccaagttgg ttgaaaccat caaccagatg caggaggaca 4680 tgtcaacaaa acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca 4740 ctggagtgaa ggctatcagg accggtctcg acgaggccaa accctggtac aagctcatca 4800 agctcttgag ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc 4860 ttgtggccat catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga 4920 agaagatctc cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg 4980 gagccccgat tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac 5040 ccgaagacct tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg 5100 ccattctcaa gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca 5160 aggcatggat cgcctcagaa gaaaagtttg tcaccatgac ggacctggtg cctggcatcc 5220 ttgaaaagca gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg 5280 acaacgcgcg ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag 5340 tggttgcccc agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca 5400 agtcaggcca gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact 5460 tcactggcag aaccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt 5520 acaaccagca gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact 5580 tcaagtactt tgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg 5640 aggacaaagg caagcctttt aacagcaagg tcatcatcgc caccaccaac ctgtactcgg 5700 gcttcacccc gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca 5760 ttgacgtgag cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc 5820 ttgaagacac ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg 5880 gcatggccgt tgaaatgaag agaatgcaac aagacatgtt caagccccaa ccgcccctcc 5940 agaacgtcta ccagcttgtt caggaggtga ttgaccgggc cgagctccac gagaaggtgt 6000 cgaaccaccc gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc 6060 tcattgagaa aggccagcac gaagcagcaa ttgaattctt tgaggggatg gtgtgtgact 6120 ccatcaagga ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta 6180 agcgcctgaa ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag 6240 tgatcatgat ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt 6300 acactgagaa ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc 6360 ctctggaaac cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca 6420 agacgggtga tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg 6480 aaggacccta caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc 6540 cgcagcagga ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag 6600 cgaaagcccc ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt 6660 tgaaggtgaa agcaaagaac ttgattgtca ctgagagcgg tgctccccct actgacttgc 6720 aaaagatggt catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg 6780 ccatctgctg cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg 6840 cagagaaata tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag 6900 tgtttgagtt tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg 6960 tgcttcaccg tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa 7020 tgaagaaagg cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct 7080 tctctggtga ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc 7140 ccggtctctt cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg 7200 caaaggacgg agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg 7260 gatactgctc atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg 7320 aaccacacca cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca 7380 tgcgcaaaac caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc 7440 ctgccgcctt gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca 7500 tcttctccaa acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc 7560 gctgtgctgc tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac 7620 tgagcattta cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg 7680 cgcctggtct cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga 7740 acggcactgt cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca 7800 agtttgtatg ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg 7860 gtaagactcg cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga 7920 ttggcagatt ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg 7980 gttgtaatcc tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg 8040 tgtgggatgt ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca 8100 tgtttgagga ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga 8160 aaactctcgt gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga 8220 tgccgtctgg ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc 8280 tctacgcctt gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct 8340 acggagacga catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc 8400 acttcaaatc ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc 8460 ttggtcactc cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa 8520 ctgggtttta caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac 8580 gtcgtgggac catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac 8640 ctgacgagta ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca 8700 gatcacttta cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc 8760 acaattggca gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac 8820 tttttcccgc ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc 8880 atgtattcta tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc 8940 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 9000 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 9060 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 9120 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 9180 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 9240 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 9300 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 9360 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 9420 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 9480 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 9540 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 9600 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 9660 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 9720 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 9780 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 9840 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 9900 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 9960 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 10020 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 10080 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 10140 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 10200 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 10260 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 10320 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 10380 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 10440 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 10500 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 10560 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 10620 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 10680 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 10740 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 10800 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 10860 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 10920 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 10980 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 11040 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11093 <210> 6 <211> 11083 <212> DNA <213> ASPs12L clone <400> 6 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 60 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 120 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 180 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 240 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 300 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 360 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 420 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 480 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 540 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 600 taaaacgacg gccagtgggc gcgcctaata cgactcacta taggttgaaa gggggcgcta 660 gggtctcatc cctaacatgc caacgacagc tcctgcgctg cactctacac ccacgtctgt 720 gtgtgtgcgg gggccgctgg actaccgtcc acccacctac ggttggactc acgacaccgc 780 gtggccattt tagctggact gtgcggacga acgccacttg cgcaccccgc gtgactggtt 840 aatactctta ccactttccg cctacttggt cgttagcgct gccctgggca ctcccgttgg 900 gggccatccg gtgctccacg gttcccacgc atgacaaact acggtgatgg ggccgcttcg 960 tgcgagttgg gcgcctggcg tgtttcggct gtcacccgaa gtccaccttt cacccccccc 1020 ccccaagttt ttaccgcctt tcccggcgtt aaagggaggt aaccacaagc ttgcgactgt 1080 cttgctcgac gataaagggc tgtgaccgca agatgacacc gcctttcccg gcgttaattg 1140 gatgtaacca caagatgaac cttcacccgg aagtaaaacg gtaaattcgc ctagttttgc 1200 ccgttttcac gagaaatggg acgtctgcgc acgaaacgcg ccgtcgcttg aggaggactt 1260 gtacaaacac ggtccattca ggttcccaca accgacacaa atcgtgcaac ttgaaactcc 1320 gcctggtctt tccaggtcta gaggggtgac attttgtact gtgcttgact ccacgctcgg 1380 tccactggcg agtactagta acagcactgc tgcttcgtag cggagcatgg tggccgcggg 1440 aattcctcct tggtaacagg gacccgcggg gccgaaagcc acgtcctcac ggacccacca 1500 tgtgtgcaac cccagcacgg caactttact gtgaaaacca ctttaaggtg acactgatac 1560 tggtactcaa tcactggtga caggctaagg atgcccttca ggtaccctga ggtaacacgc 1620 gacactcggg atctgagaag gggactgggg cttctttaaa agcgcctagt ttaaaaagct 1680 tctacgcctg aataggtgac cggaggccgg cacctttctt ttaaacaact atttttaaat 1740 gagcacaacc gactgtttca tcgctttgtt gtacgccttc agagagatta aaacactgtt 1800 cttatcacaa gcacaaggaa agatggagtt cacacttcac aacggtgaga agaaaacatt 1860 ctactccagg cccaacaacc acgacaactg ctggctgaac gccatcctcc agttgtttag 1920 gtacgttgat gaacctttct tcgactgggt ctactactca cctgagaacc tcacgctcga 1980 tgctatcaaa caattggaag gaattactgg tctcgagctc cacgagggtg gaccacccgc 2040 tctcgttatt tggaacatta aacacctgct caacaccgga atcggcaccg cttcgcgacc 2100 cagcgaagtg tgcatggtag atgggacgga catgtgtttg gctgacttcc acgctggcat 2160 cttcctgaaa ggacaggaac acgctgtgtt cgcctgtgtc acctccaacg ggtggtacgc 2220 gattgatgac gaggactttt acccctggac gccggacccg tctgacgttc tggtgttcgt 2280 cccgtacgac caagaaccgc tcaacggaga atggaaagca aaggttcaga aacgactcag 2340 aggcgccggg caatccagcc cggcgactgg gtcgcagaac cagtcaggta acactggaag 2400 cattatcaac aattactaca tgcagcagta ccagaactcc atggacacac aacttggtga 2460 caacgctatt agcggaggct ccaacgaggg gtccacggac accacctcca cccacacaac 2520 caacactcag aataatgact ggttttcaaa gctggccagt tccgctttta gcggtctttt 2580 cggcgctctc ctcgccgaca agaaaaccga ggagaccact cttctcgagg accgcatcct 2640 cactacccgc aacggacaca cgacctcgac aacccagtcg agcgttggag tcacttacgg 2700 gtacgcaaca gctgaggact tcgtgagcgg accaaacaca tctgggcttg agaccagggt 2760 tgtgcaggca gagcggttct ttaaaaccca cttgttcgac tgggtcacca gtgacccgtt 2820 cggacggtgc tacctgctgg aactcccaac tgaccacaaa ggtgtctacg gcagcctgac 2880 cgactcttat gcttacatga gaaacggttg ggacgttgag gtcactgcgg tgggaaatca 2940 gttcaacgga ggatgcttgt tggtggccat ggtgccagaa cattgctcta ttgacaagag 3000 agggctgtac cagctcacgc tctttcccca ccagttcatc aacccccgga cgaacatgac 3060 ggcgcacatt actgtgccct ttgttggcgt caaccgctac gaccagtaca aagtacacaa 3120 accttggacc ctcgttgtca tggttgtggc cccgctgact gtcaacaccg aaggtgcccc 3180 acagatcaag gtctatgcca acatcgcccc tactaacgtg cacgttgcgg gtgagctccc 3240 ttctaaggaa gggatcttcc ccgtggcatg tagcgacggt tacggtggtc tggtgaccac 3300 tgacccaaag acggctgacc ccgcctacgg gaaagtgttc aatccacctc gcaacatgtt 3360 gccggggcgg ttcaccaact tccttgatgt ggctgaggcg tgccctacgt tcctgcactt 3420 tgagggtgac gtgccgtacg tgaccacaaa gacggactca gacagggtac tcgcccagtt 3480 tgacttgtct ctggcagcaa agcacatgtc aaacaccttc ctggcaggtc tcgcccagta 3540 ctacacacag tacagcggca ccatcaacct gcacttcatg ttcacaggac ccactgacgc 3600 gaaagcgcgt tacatgattg catacgcccc ccctggcatg gagccgccca aaacacccga 3660 ggcggccgct cactgcattc acgcggagtg ggacacaggg ttgaattcaa aattcacatt 3720 ttcaatccct tacctttcgg cggctgatta tgcgtacacc gcgtctgacg ccgcggagac 3780 cacaaatgtg cagggttggg tttgcctgtt tcaaattaca cacgggaagg ctgacggcga 3840 cgcactggtc gttctagcta gcgccggtaa ggactttgaa ctgcgtctgc cagttgacgc 3900 tcgcacgcag accacctcca caggtgagtc ggctgacccc gtgactgcca ccgttgagaa 3960 ctacggtggt gagacacagg tccagagacg ccaacacacg gatgtctcgc tcatactaga 4020 cagatttgtg aaagtaacac caaaagacca aattaatgtg ttggacctga tgcaaatccc 4080 tgcacacact ttggtaggcg cgctcctccg tactgccacc tactacttcg cagatctgga 4140 agtggcagtg aaacacgagg ggaacctcac ctgggtcccg aacggggcgc ccgaggcagc 4200 gttggacaac accaccaatc caacggccta tcacaaggcg ccgctcaccc ggcttgcact 4260 gccttacacg gcaccacacc gtgtcttggc tactgtttac aacgggaact gcaagtatgg 4320 caagagcccc gtgaccaatc tgagaggtga cctgcaagtg ttgacccaga aggcggcaag 4380 aacgctgcct acctccttca attacggtgc catcaaagcc actcgggtga ctgaactgct 4440 ttaccgcatg aagagggccg aaacatactg cccccggcct cttttggcta ttcacccgag 4500 cgaagctaga cacaaacaaa agattgtggc gcctgtgaaa cagctgttga actttgacct 4560 gctcaagttg gcaggagacg tcgagtccaa ccctgggccc ttcttcttct ctgacgtcag 4620 gtcaaatttt tccaagttgg ttgaaaccat caaccagatg caggaggaca tgtcaacaaa 4680 acacggaccc gactttaacc ggttggtgtc tgcatttgag gaactggcca ctggagtgaa 4740 ggctatcagg accggtctcg acgaggccaa accatggtac aagctcatca agctcttgag 4800 ccgcctgtca tgtatggccg ctgtagcagc acggtcaaag gacccactcc ttgtggccat 4860 catgctggct gacaccggcc ttgagatcct ggacagtacc tttgtcgtga agaagatctc 4920 cgactcgctc tccagtctct ttcacgtgcc ggcccccgtc ttcagtttcg gagccccgat 4980 tttgttggcc gggttggtca aagtcgcctc gagtttcttc cggtccacac ccgaagacct 5040 tgagagagcg gagaaacagc tcaaagcacg tgacatcaat gacatattcg ccattctcaa 5100 gaacggcgag tggctggtca agctgattct tgccatccgc gactggatca aggcatggat 5160 cgcctcagaa gaaaagtttg tcaccatgac agacctagtg cctggcatcc ttgaaaagca 5220 gcgggatctc aacgacccaa gcaagtacaa ggaggccaag gagtggctcg acaacgcgcg 5280 ccaagcgtgt ctgaagagcg ggaacatcca catcgcaaac ctttgcaaag tggttgcccc 5340 agcgcccagc aggtcgaggc ccgaacccgt ggtcgtttgc ctccgtggca agtcaggcca 5400 gggcaagagt ttccttgcga acgtgcttgc acaagcaatt tcaacccact tcactggcag 5460 aaccgattca gtttggtact gcccacctga ccctgaccac ttcgacggtt acaaccagca 5520 gaccgttgta gtaatggatg atttgggcca gaaccccgac gggaaggact tcaagtactt 5580 cgcccaaatg gtttcaacta cggggtttat cccgcccatg gcttcactcg aggacaaagg 5640 caagcctttt aacagcaagg tcatcatcgc caccaccaac ctgtactcgg gcttcacccc 5700 gagaactatg gtgtgccctg atgcactgaa ccgaaggttc cactttgaca ttgacgtgag 5760 cgccaaggac gggtacaaaa ttaacaacaa attggacatc atcaaagctc ttgaagacac 5820 ccacaccaac ccagtggcaa tgtttcaata cgactgtgcc cttctcaacg gcatggccgt 5880 tgaaatgaag agaatgcaac aagacatgtt caagcctcaa ccgcccctcc agaacgtcta 5940 ccagcttgtt caggaggtga ttgaccgggt cgagctccac gagaaggtgt cgaaccaccc 6000 gattttcaag cagatctcaa ttccttccca aaaggctgtg ctgtactttc tcattgagaa 6060 aggccagcac gaagcagcaa ttgaattctt tgaggggatg gtgtgtgact ccatcaagga 6120 ggagctccgg cctctcatcc aacagacctc atttgtgaag cgcgctttta agcgcctgaa 6180 ggaaaacttt gagatagttg ccctgtgttt gactctaatg gcaaacatag tgatcatgat 6240 ccgcgagact cgcaagagac agcagatggt ggatgatgca gtgaacgagt acactgagaa 6300 ggtaaacatc accacggatg acaagactct tgacgaggcg gaaaagaacc ctctggaaac 6360 cagcggtgcc accactgttg gtttcagaga gaaaactctc ccggggcaca agacgggtga 6420 tgacgtgagc tccgagccca ccaaacccgt ggaaggacaa ccacaagctg aaggacccta 6480 caccggtcca ctcgagcgtc aaaaacctct gaaagtgaga gccaagctcc cgcagcagga 6540 ggggccctat gctggcccca tggagagaca gaaaccactg aaagtgaaag cgaaagcccc 6600 ggtcgttaag gaaggacctt acgaaggacc ggtaaagaaa cctgtcgctt tgaaggtgaa 6660 agcaaagaac ttgattgtca ctgagagtgg tgctccccct actgacttgc aaaagatggt 6720 catgggtaac accaagcctg ttgagctcgt cctcgacggg aagacggtgg ccatctgctg 6780 cgccaccgga gtgtttggta ctgcctacct tgttcctcgt catcttttcg cagagaaata 6840 tgacaagatc atgctggacg gcagagccat gacagacagt gactacagag tgtttgagtt 6900 tgagattaaa gtgaaaggac aggacatgct ctcagacgcc gcgctcatgg tgcttcaccg 6960 tgggaatcga gtgcgggaca tcacgaagca cttccgtgat gtggcaagaa tgaagaaagg 7020 cacccccgtc gtcggcgtga tcaacaacgc tgatgttggg aggctgatct tctctggtga 7080 ggcccttacc tacaaggaca ttgtagtgtg catggacgga gacaccatgc ccggtctctt 7140 cgcctacaaa gccgccacca aggcgggcta ctgtggagga gccgttcttg caaaggacgg 7200 agccgagact ttcatcgtcg gcactcactc cgcaggcggc aatggagttg gatactgctc 7260 atgcgtttcc aggtctatgc tgcttaaaat gaaggcacac atcgatcccg aaccacacca 7320 cgagggattg atagttgaca ccagagatgt tgaggagcgc gtacacgtca tgcgcaaaac 7380 caagctcgca cccaccgtag catacggtgt gtttaacccc gaatttgggc ctgccgcctt 7440 gtccaacaag gacccgcgcc tgagtgaggg ggttgtcctc gatgaagtca tcttctccaa 7500 acacaaagga aacacaaaga tgtctgagga ggacagagcg ctgttccgcc gctgtgctgc 7560 tgactacgcg tcgcgtctgc acagtgtgct gggtacggca aatgccccac tgagcattta 7620 cgaggcaatc aagggcgtcg acggacttga cgccatggaa ccagacaccg cgcctggtct 7680 cccctgggct ctccagggga aacgccgtgg tgcgctcatt gacttcgaga acggcactgt 7740 cggacccgag gttgaagctg ccttgaagct catggagaaa agagagtaca agtttgtatg 7800 ccagaccttc ttgaaggacg agattcgccc gatggagaag gtacgtgccg gtaagactcg 7860 cattgtcgac gtcctgcctg ttgaacacat tctttacacc aggatgatga ttggcagatt 7920 ttgtgctcaa atgcactcaa acaacggacc gcaaattggc tcggcggttg gttgtaatcc 7980 tgatgttgat tggcaaagat ttggcacgca ctttgctcag tacagaaacg tgtgggatgt 8040 ggactattcg gccttcgacg ccaaccactg cagtgacgca atgaacatca tgtttgagga 8100 ggtgttcaac acggatttcg gtttccaccc aaacgctgag tggatcctga aaactctcgt 8160 gaacactgaa cacgcctacg agaacaaacg catcactgtt gaaggcggga tgccgtctgg 8220 ttgttccgcg acaagcatca tcaacacaat tttgaacaac atctacgtgc tctacgcctt 8280 gcgtagacac tacgagggag ttgagctgga ctcttacacc atgatctcct acggagacga 8340 catcgtggtt gcaagtgatc acgatctgga ctttgaggcc ctcaagcctc acttcaaatc 8400 ccttggtcaa accatcactc cagctgacaa aagcgacaaa ggttttgttc ttggtcactc 8460 cattaccgat gtcactttcc tcaaaagaca cttccacatg gactatggaa ctgggtttta 8520 caaacctgtg atggcttcga agaccctcga ggctatcctc tcctttgcac gtcgtgggac 8580 catacaggag aagttgatct ccgtggcagg actcgccgtc cactctggac ctgacgagta 8640 ccggcgtctc tttgagcctt tccagggtct ctttgagatt ccaagctaca gatcacttta 8700 cctgcgttgg gtgaacgccg tgtgcggtga cgcataatcc ctcagatgtc acaattggca 8760 gaaagacgct gaggcgagcg acgccgtagg agtgaaaagt ccgaaaggac tttttcccgc 8820 ttcctatttc aagagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgc atgtattcta 8880 tagtgtcacc taaatcatat gggagagctc ccaacgcgct tggcgtaatc atggtcatag 8940 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 9000 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 9060 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 9120 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 9180 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 9240 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 9300 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 9360 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 9420 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 9480 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 9540 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 9600 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 9660 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 9720 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 9780 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 9840 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 9900 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 9960 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 10020 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 10080 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 10140 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 10200 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 10260 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 10320 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 10380 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 10440 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 10500 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 10560 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 10620 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 10680 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 10740 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 10800 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 10860 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 10920 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 10980 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 11040 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac 11083

Claims (7)

서열번호 1과 같은 염기서열을 갖는 구제역바이러스를 이용하여 작성한 cDNA 클론(clone).
CDNA clone prepared using foot-and-mouth virus having the same nucleotide sequence as SEQ ID NO: 1.
서열번호 2와 같은 염기서열, 서열번호 3와 같은 염기서열, 서열번호 4와 같은 염기서열 및 서열번호 5와 같은 염기서열로 이루어진 군에서 선택된 어느 하나의 염기서열을 갖는 구제역바이러스를 이용하여 작성한 클론.
Clones created using foot-and-mouth virus having any one nucleotide sequence selected from the group consisting of a nucleotide sequence such as SEQ ID NO: 2, a nucleotide sequence such as SEQ ID NO: 3, a nucleotide sequence such as SEQ ID NO: 4, and a nucleotide sequence such as SEQ ID NO: 5 .
서열번호 5와 같은 염기서열을 갖는 클론의 UTR(untranslated region) 또는 T7 promotor 부위를 수정하여 replicon의 복제 및 발현의 차이를 확인하게 한 클론으로서, T7 promotor 부위 644nt G 뒤에 G가 하나 더 삽입되고, 9602nt 위치의 C가 T로 치환된 것을 특징으로 하는 클론.
A clone that modifies the untranslated region (UTR) or the T7 promotor region of a clone having a nucleotide sequence such as SEQ ID NO: 5 to identify the difference in replication and expression of the replicon, wherein one more G is inserted after the T7 promotor region 644nt G. C at position 9602nt is substituted with T. Clone.
서열번호 5와 같은 염기서열을 갖는 클론의 UTR 또는 T7 promotor 부위를 수정하여 replicon의 복제 및 발현의 차이를 확인하게 한 클론으로서, 1309nt 위치에서 A가 G로 치환되고, 9458nt 위치의 C가 A로 치환된 것을 특징으로 하는 클론.
A clone that modifies the UTR or T7 promotor region of a clone with a nucleotide sequence such as SEQ ID NO: 5 to identify the difference in replication and expression of the replicon. Clone, characterized in that substituted.
서열번호 5와 같은 염기서열을 갖는 클론의 UTR 또는 T7 promotor 부위를 수정하여 replicon의 복제 및 발현의 차이를 확인하게 한 클론으로서, 1309nt 위치의 A가 G로 치환되고, 9602nt 위치의 C가 T로 치환된 것을 특징으로 하는 클론.
A clone that modified the UTR or T7 promotor region of a clone having a nucleotide sequence as shown in SEQ ID NO: 5 to identify a difference in replication and expression of replicon, wherein A at 1309nt was replaced with G, and C at 9602nt was replaced with T. Clone, characterized in that substituted.
서열번호 6과 같은 염기서열을 갖는 replicon 클론.
Replicon clone having the same nucleotide sequence as SEQ ID NO: 6.
제6항의 클론에서 poly(C) 부위를 12개에서 24~36개로 연장한 것을 특징으로 하는 감염성 클론.The infectious clone of claim 6, wherein the poly (C) region extends from 12 to 24 to 36 in the clone of claim 6.
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20190032299A (en) * 2017-08-01 2019-03-27 지프로안 바이오테크놀로지즈 (스저우), 아이엔씨. Foot-and-mouth disease virus-like particle vaccine and method for its production
WO2021169768A1 (en) * 2020-02-25 2021-09-02 中国农业科学院兰州兽医研究所 Method for in vitro assembly of virus-like particles of foot-and-mouth disease and use thereof
KR20220106945A (en) * 2020-06-25 2022-08-01 대한민국(농림축산식품부 농림축산검역본부장) Novel adjuvant, and vaccine composition comprising the same
CN117771354A (en) * 2024-02-23 2024-03-29 云南省畜牧兽医科学院 mRNA-LNP vaccine for preventing O-type foot-and-mouth disease and preparation method thereof

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20190032299A (en) * 2017-08-01 2019-03-27 지프로안 바이오테크놀로지즈 (스저우), 아이엔씨. Foot-and-mouth disease virus-like particle vaccine and method for its production
WO2021169768A1 (en) * 2020-02-25 2021-09-02 中国农业科学院兰州兽医研究所 Method for in vitro assembly of virus-like particles of foot-and-mouth disease and use thereof
KR20220106945A (en) * 2020-06-25 2022-08-01 대한민국(농림축산식품부 농림축산검역본부장) Novel adjuvant, and vaccine composition comprising the same
CN117771354A (en) * 2024-02-23 2024-03-29 云南省畜牧兽医科学院 mRNA-LNP vaccine for preventing O-type foot-and-mouth disease and preparation method thereof

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