KR20160003691A - Opa1 단상부족에 의해 원인이 되는 질병 치료를 위한 인공전사인자 - Google Patents

Opa1 단상부족에 의해 원인이 되는 질병 치료를 위한 인공전사인자 Download PDF

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KR20160003691A
KR20160003691A KR1020157031596A KR20157031596A KR20160003691A KR 20160003691 A KR20160003691 A KR 20160003691A KR 1020157031596 A KR1020157031596 A KR 1020157031596A KR 20157031596 A KR20157031596 A KR 20157031596A KR 20160003691 A KR20160003691 A KR 20160003691A
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알버트 네우트체트너
조세프 플래머
앨리스 헉슬리
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Abstract

본 발명은 활성 단백질 도메인(activatory protein domain), 및 핵국재화서열(nuclear localization sequence)에 융합된 OPA1 프로모터를 특정적으로 표적하는 폴리닥틸 징크핑거 단백질(polydactyl zinc finger protein)을 포함하는 인공전사인자에 관한 것이다. OPA1 프로모터에 대하여 직접적인 인공전사인자(transcription factors)는 상염색체성우성시신경위축(autosomal dominant optic atrophy), 상염색체성우성시신경위축 증후군 플러스(syndromic autosomal dominant optic atrophy plus) 및 정상 긴장상태 녹내장(normal tension glaucoma)과 같은 OPA1단상부족(haploinsufficiency)과 연관된 질병 치료에 유용하다.

Description

OPA1 단상부족에 의해 원인이 되는 질병 치료를 위한 인공전사인자{Artificial transcription factors for the treatment of diseases caused by OPA1 haploinsufficiency}
본 발명은 활성도메인(activatory domain) 및 핵국재화서열(nuclear localization sequence)에 융합된 특정적으로 OPA1 유전자 프로모터를 표적하는 폴리닥틸 징크핑거단백질(polydactyl zinc finger protein), 및 단상부족(haploinsufficiency)을 유도하는 OPA1에서 돌연변이에 의해 원인이 되는, 상염색체성 우성 시신경위축(autosomal dominant optic atrophy) (ADOA) 또는 증후군 ADOA 플러스와 같은 질병 치료에서 이들의 용도를 포함하는 인공전사인자(transcription factors)에 관한 것이다.
인공전사인자(transcription factors) (ATFs)는 유전자 발현을 조절하는 유용한 도구로서 제안되었다(Sera T., 2009, Adv Drug Deliv Rev 61, 513-526). 유전자 발현의 억제(repression) 또는 활성을 통하여 유전자 발현에 영향을 주는, 많은 자연적으로 발생하는 전사인자(transcription factors)는 확실한 DNA 서열의 인식을 위하여 복합 특정 도메인(complex specific domains)을 소유한다. 만약 누군가 그들의 특수성 및 표적유전자를 수정하기를 의도한다면 이것은 그즐을 조작을 위하여 흥미없는 표적으로 만든다. 그러나, 전사인자(transcription factors)의 확실한 분류는 모듈(modular) 및 유전자조작을 그들 스스로에 빌려주는, 징크 핑거(ZF) 도메인으로 불리는, 몇 가지를 함유한다. 징크핑거는 거의 비의존적 3개의 DNA 베이스 쌍(base pairs)을 표적하는 짧은(30 아미노산) DNA 결합 모티프이다. 따라서 함께 융합된 몇 개의 상기 징크 핑거를 포함하는 단백질은 긴 DNA 서열을 인식할 수 있다. 헥사메릭징크핑거단백질(hexameric zinc finger protein)(ZFP)는 전체 인간게놈에서 거의 고유한, 18 베이스쌍(base pairs) (bp) DNA 표적을 인식한다. 초기에 완전히 상황(context)에 비의존적으로 여겼던, 더욱 인-뎁스(in-depth) 분석은 징크핑거에 대한 일부 상황(context) 특수성을 드러냈다(Klug A., 2010, Annu Rev Biochem 79, 213-231). ZF 모듈의 결합특수성을 변형하는 징크핑거 인식표면에서 특정 아미노산의 돌연변이는 5’-GNN-3’, 5’-CNN-3’, 5’-ANN-3’의 대부분, 및 일부 5’-TNN-3’ 코돈 (예들 들면, 소위 Barbas 모듈, Dreier B., Barbas C.F. 3rdet al., 2005, J Biol Chem 280, 35588-35597을 참조하시오)에 대하여 정의된 ZF 빌딩 블록(ZF building blocks)을 초래한다. 인공전사인자(transcription factors)에 대해 초기 작업은 알려진 3 bp 표적서열로 된 미리 선별된 징크핑거 혼합에 기반한 이성적 디자인에 집중된 반면에, 징크핑거의 확실한 상황(context) 특수성의 실현은 FACS 분석을 사용하여 박테리아 또는 이스트원하이브리드(yeast one hybrid), 파지 디스플레이(phage display), 구분된 리보좀 디스플레이(ribosome display) 또는 생체내(in vivo)선별과 같은 복잡한 방법을 사용하여 조사되는, 큰 징크핑거 라이브러리의 발생을 필요로 한다.
상기 인공 징크핑거 단백질을 사용하여, 인간게놈 내에 DNA 유전자 자리(loci)는 높은 특수성으로 표적될 수 있다. 따라서, 상기 징크핑거 단백질은 특정한 프로모터 서열에 전사-조절(modulatory)활성을 가지는 단백질 도메인을 운반(transport) 하는데 이상적 도구이고 이는 관심있는 유전자 발현 조절을 초래한다. 유전자 전사의 활성을 위한 적절한 도메인은 헤르페스 심플렉스 바이러스 VP16(herpes virus simplex VP16)(서열번호:1) 또는 P64 (VP16의 4분자체(tetrameric) 반복, 서열번호:2) 도메인(Beerli R.R. et al., 1998, Proc Natl Acad Sci USA 95, 14628-14633)이다. 전사 활성을 부여하는 것으로 고려되는 추가 도메인은 CJ7(서열번호:3), p65-TA1(서열번호:4), SAD(서열번호:5), NF-1(서열번호:6), AP-2(서열번호:7), SP1-A(서열번호:8), SP1-B(서열번호:9), Oct-1(서열번호:10), Oct-2 (서열번호:11), Oct-2_5x (서열번호:12), MTF-1(서열번호:13), BTEB-2(서열번호:14) 및 LKLF(서열번호:15)이다. 추가적으로, 유전자 존재론(ontology) GO:0001071 (http://amigo.gene존재론(ontology).org/cgi bin/amigo/term_details?term=GO:0001071)에 의해 정의되는 단백질의 전사적으로활성 도메인은 표적단백질의 전사조절을 얻는것으로 고려된다.
소분자 약물은 특정한 특색의 높은 보존에 기인하여 주어진 단백질 페밀리의 확실한 구성원을 선별적으로 표적할 수 없는 반면에, 생물학적제제(biologicals)는 항체-기반 신규약물에 대하여 나타낸 바와 같이 생물학적제제(biologicals)는 큰 특수성을 제안한다. 그러나, 사실상 지금까지 모든 생물은 세포밖으로 활동한다. 특히 상기 언급된 인공전사인자(transcription factors)는 치료적으로 유용한 방식으로 유전자 전사를 영향주는데 적절할 것이다. 그러나, 상기 작용 부위-핵(nucleus)-에 상기 인자를 운반하는 것은 쉽게 얻어지지 않아서, 따라서 예들 들면, 면역원성(immunogenicity) 및 세포의 변형을 위한 잠재성과 같은 상기 방법의 모든 결점을 가지는 레트로바이러스 운반에 의존한, 치료 인공 전사인자 접근의 유용성을 방해한다(Lund C.V. et al., 2005, Mol Cell Biol 25, 9082-9091).
소위 단백질 형질도입(transduction) domains(PTDs))은 원형질막(plasma membrane)을 가로질러 세포질/핵원형질(cytosol/nucleoplasm)로 단백질 위치변경(translocation)을 촉진하는 것으로 나타난다. HIV 유도된 TAT 펩타이드 (서열번호:16) 및 그박의 것과 같은 짧은 펩타이드는 카고(cargo)단백질의 세포-유형 비의존적 거대 음작용 섭취(macropinocytotic uptake)를 유도하는 것으로 나타낸다(Wadia J.S. et al., 2004, Nat Med 10, 310-315). 사이토졸(cytosol)에 도착에 따라, 상기 융합(fusion)단백질은 생물활성을 가지는 것으로 나타난다. 흥미롭게도, 미스폴드된(misfolded) 단백질은 하기의 세포 내의(intracellular) 샤페론의 작용을 통하여 기능적이 될 수 있다.
유전자 돌연변이는 많은 유전적 질병의 심장에 있다. 일반적으로, 질병 모두에 원인이 되는 열성(recessive) 돌연변이에 관하여, 어머니 같은(maternal) 및 아버지 같은(paternal), 유전자 복사(copy)가 돌연변이 되는 것이 필요한 반면에, 상기 돌연변이는 심지어 단지 하나의 유전자 복사(copy)-어머니 같은(maternal) 또는 아버지 같은(paternal)-영향받을 때 질병형태의 원인이 되는 우성 돌연변이를 가지는, 유전적(inheritance) 모드에 관하여 우성(dominant) 또는 열성(recessive)으로 분류될 수 있다. 우성 돌연변이는 도미넌트-네거티브(dominant-negative) 작용 또는 단상부족(haploinsufficiency)에 의해, 2개의 일반적 기전의 하나에 의해 질병을 일으킬 수 있다. 도미넌트-네거티브(dominant-negative)돌연변이의 경우에, 유전자 생성물은 새로운, 독성이 있고 질병형태의 원인이 되는 비정상 기능을 얻는다. 실시예는 상기 돌연변이가 단백질 복합체의 적절한 기능을 방지하는 멀티메릭(multimeric) 단백질 복합체의 서브유닛이다. 또한 우성 패션으로 유전된 질병은 단상부족(haploinsufficiency)에 의해 원인이 될 수 있고, 상기 돌연변이 원인이 되는- 질병은 영향 받은 유전자를 비활성화하고, 따라서 효과적 유전자 용량을 낮춘다. 이러한 환경하에서, 두번째, 온전한 유전자 복사(copy)는 정상 기능을 위하여 충분한 유전자 생성물을 제공할 수 없다. 약 12’000 인간 유전자는 질병과 연관되는 것으로 알려진 약 300 유전자를 가지는 단상부족(haploinsufficiency)인 것으로 추정된다(Huang et al., 2010, PLoS Genet. 6(10), e1001154).
신경 생존성은 많은 신경병성 장애의 심장에서 미토콘드리아 불이행을 가지는 미토콘드리아 기능에 임계적으로 의존한다(Karbowski M., Neutzner A., 2012, Acta Neuropathol 123(2),157-71). ATP 형성에서 에너지를 제공하는 것에서 그들의 필수 기능 이외에, 미토콘드리아는 대사작용(metabolic) 과정 및 또한 세포사멸(programmed cell death) 뿐만 아니라 칼슘 버퍼링, 다양한 이화작용에 임계적으로 관련된다. 미토콘드리아의 이러한 중요한 기능은 미토콘드리아를 유지하고 미토콘드리아 불이행 및 그 뒤에 세포죽음(cell death)을 방지하는 적소에서 많은 세포의 기전으로 비춰진다(Neutzner A. et al., 2012, Semin Cell Dev Biol 23, 499-508). 상기 과정 중에 중심 역할은 균형된 미토콘드리아 형태론을 가지는 다이나믹 미토콘드리아 네트위크의 유지를 수행한다. 이것은 Drp1, Fis1, Mff, MiD49 및 MiD51 - Mfn1, Mfn2 및 OPA1 경우에 미토콘드리아 튜불(tubules)의 융합의 경우에서 미토콘드리아의 핵분열(fission)을 촉진하는 소위 미토콘드리아 모르포겐(morphogens)에 의해 얻어진다. 미토콘드리아 융합(fusion) 결실이 ATP 생성물의 결실을 촉진하고 신경병성 장애와 연관되는 신경 세포죽음(neuronal cell death)에 상기 과정을 연결하는 세포사멸(apoptotic) 자극에 세포를 민감시키는 것으로 알려져 있기 때문에 미토콘드리아 형태를 균형하는 것은 필수적이다.
미토콘드리아 융합(fusion)의 과정에서 주요 인자는 시신경 위축(optic atrophy) 1 또는 OPA1이다. OPA1은 OPA1 유전자에 의해 암호화 되는 큰 GTPase이고 미토콘드리아 융합에 필수적이다. 추가적으로, OPA1은 크리스태(cristae)의 구성요소로서 내부, 미토콘드리아 구조를 유지하는데 중요한 역할을 한다. OPA1유전자 발현의 하향조절(downregulation)은 융합 결실에 기인하여 미토콘드리아 단편을 일으키고 세포사멸(apoptotic) 자극에 세포를 민감하게 한다.
OPA1에서 돌연변이는 Kjer의 시각 신경병증(optic neuropathy) 또는 보통염색체 우성 위축증(autosomal dominant atrophy) (ADOA)의 약 70%에 대하여 책임있는 것으로 확인되었다. 대부분의 인구에서, ADOA는 1/10’000와 3/100’000사이에서 유행하고 유아기(early childhood)에서 시작하는 시각 감소를 천천히 진행하는 것으로 특징된다. 약한 것으로부터 법률적으로 맹인(blind)까지의 범위의 시각 손상은 비가역적(irreversible)이고, 망막 신경절(retinal ganglion)세포(RGCs)의 느린 퇴화에 의한 원인이 된다. 대부분의 경우에서, ADOA는 비-증후군(non-syndromic)이지만, 눈-외의(extra-ocular) 환자의 약 15%에서, 감각-뇌 청각(sensori-neural hearing)결실과 같은 신경근 징후(neuro-muscular manifestation)을 만난다. 현재까지, 상기 질병을 위한 보이는 치료가 사용가능하지 않다. 흥미롭게도, 확실한 OPA1 대립형질(alleles)은 정상 긴장상태(tension)에 연결되지만, 높은 긴장상태(tension)에는 연결되지 않고, 정상 미토콘드리아 생리(physiology)를 유지하는데 OPA1의 중요성을 다시 하이라이트한다.
도 1: 변환 가능한 인공전사인자( transducible transcription factors)를 사용하여 단상부족(haploinsufficiency)을 완화하기 위한 치료 접근
(A) 단상부족(haploinsufficiency) 돌연변이 (HM)는 야생형 상황(WT)과 비교하여 프로모터(P)의 대조군 하에서 유전자(G)로부터 유전자 생성물 발생(GP)의 감소를 일으킨다.
(B) 핵국재화서열(nuclear localization sequence) (NLS) 뿐만 아니라 활성도메인(활성 domain) (RD)에 융합된 단상부족(haploinsufficiency) 유전자(G)의 프로모터(P) 부위를 특정적으로 표적하는 헥사메릭징크핑거(hexameric zinc finger) (ZF) 단백질을 포함하는 인공전사인자는 TAT 또는 그 밖의 것과 같은 단백질 형질도입(transduction) 도메인 (PTD)의 작용에 의해 세포로 운반된다. 돌연변이된 (HM) 및 야생형유전자(G)에 결합에 따라, 야생형 유전자 복사(copy)로부터 유전자 생성물의 생성은 돌연변이된 유전자 복사(copy)로부터 유전자 생성물의 결실에 대하여 대체하는 것으로 증가된다.
(C) 핵국재화서열(nuclear localization sequence) (NLS) 뿐만 아니라 활성도메인에 융합된 단상부족(haploinsufficiency)유전자(G)의 프로모터(P) 부위를 특정적으로 표적하는 헥사메릭징크핑거(hexameric zinc finger) (ZF)를 포함하는 인공전사인자는 상기 인공전사인자에 대하여 cDNA 코딩의 바이러스 형질도입(transduction)에 따른 세포에 의해 발현된다. 돌연변이된 (HM) 및 야생형유전자(G)의 프로모터에 결합에 따라, 야생형 유전자복사(copy)로부터 유전자 생성물의 생성은 돌연변이된 유전자 복사로부터 유전자 생성물의 결실에 대하여 대체하는 것으로 증가된다.
도 2: OPA1 프로모터 부위
나타낸 OPA1 프로모터부위는 PA1 프로모터(서열번호:17)를 포함하는 OPA1의 5’ 비독해부위이다. 하이라이트된 부분은 본 발명의 인공전사인자를위한 결합위치이다(밑줄친 부분(underlined), 85부터 102까지 부위 및 91부터 108까지 위치, 834부터 853까지 위치, 및 983부터 1000까지 위치 겹치는 부분), 및 전사 시작 (굵은 부분(bold))을 위한 846 위치.
도 3: OPA1 -특정한 인공전사인자의 활성을 평가하기 위한 루시퍼리제 리포터 분석
HeLa세포를 OPA1_akt1 내지 OPA1_akt5 (panel A, 라벨된 A1 내지 A5) 또는 OPA1_akt6 내지 OPA1_akt10 (panel B, 라벨된 A6 내지 A10) 및 인간 OPA1 프로모터의 대조군 하에서 가우시아(Gaussia) 루시퍼라제를 및 CMV 프로모터의 대조군 하에서 배출된 알칼린 포스파타제를 포함하는 리포터 플라스미드를 위한 발현 플라스미드로 공동-형질감염되었다. 비활성(수정된) OPA1_akt1 (panel A) 또는 비활성(수정된) OPA1_akt6 (panel B)로 형질감염, 여기서 징크핑거 단백질에 있는 모든 징크-동등한 시스테인 잔기는 세린(serine) 잔기로 교환되고, 대조군 (라벨 C)으로서 수행된다. 루시퍼라제 및 배출된 알칼린 포스파타제 활성은 공동-형질감염후 48시간에 측정되었다. 루시퍼라제 활성을 배출된 알칼린 포스파타제 활성으로 정상화(normlized)시켰고 대조군의 퍼센트로서 발현시켰다(상대적 루시퍼라제 활성 - RLA). SD를 묘사하는 에러 바(error bars)를 가지는 평균 3개의 비의존적 실험을 나타낸다.
발명의 요약
활성 단백질 도메인 및 핵국재화서열에 융합된 OPA1 프로모터를 표적하는 폴리닥틸 징크핑거 단백질(polydactyl zinc finger protein)을 포함하는 인공전사인자, 및 상기 인공전사인자를 포함하는 약학적 조성물에 관한 것이다.
추가적으로, 본 발명은 활성 단백질 도메인, 핵국재화서열(nuclear localization sequence) 및 단백질 형질도입(transduction) 도메인에 융합된 OPA1 프로모터를 표적하는 폴리닥틸 징크핑거 단백질(polydactyl zinc finger protein)을 포함하는 인공전사인자, 및 인공전사인자를 포함하는 약학적 조성물에 관한 것이다.
또한 본 발명은 OPA1 유전자의 발현을 증강시키고 OPA1 유전자 생성물의 발생을 개선시키는 인공전사인자(artificial transcription factors)의 용도에 관한 것이다.
추가적으로, 본 발명은 낮은 OPA1 수준에 의해 원인이 되거나 수정되는 질병 치료에서 인공전사인자의 용도, 특히 ADOA 및 ADOA 플러스와 같은 안구 질병 치료의 용도에 관한 것이다. 이와 같이 본 발명은 이를 필요로 하는 환자에게 발명의 인공전사인자의 치료학적으로 유효량을 투여하는 것을 포함하는 낮은 OPA1 수준에 의해 영향받는 질병을 치료하는 방법에 관한 것이다.
본 발명의 상세한 설명
본 발명은 활성 단백질 도메인, 핵국재화서열(nuclear localization sequence) (NLS), 및 선택적으로 단백질형질도입(transduction)도메인 (PTD)에 융합된 OPA1 프로모터(서열번호:17)에 특정적으로 표적하는 폴리닥틸 징크핑거 단백질(polydactyl zinc fonger protein)(ZFP)을 포함하는 인공전사인자(ATF), 및 상기 인공전사인자를 포함하는 약학적 조성물에 관한 것이다(도 1).
본 발명의 상황에서, 프로모터는 유전자의 활성 부위(regulatory region)으로서 정의된다. 본 정의는 당업계에서 일반적 정의에 해당된다. 또한 본 발명의 상황에서, 단지 만약 2개의 기능적 유전자 복사가 게놈에서 존재한다면 단상부족(haploinsufficiency) 프로모터는 모든 경우하에서 모든 세포 유형에서 충분한 유전자 생성물의 생성을 일으킬 수 있는 프로모터로서 정의된다. 따라서, 단상부족(haploinsufficiency)유전자의 하나의 유전자 복사의 돌연변이는 일부 또는 모든 생리적 상황하에서 유기체(organism)의 일부 또는 모든 세포에서 충분한 유전자 생성물 발생을 일으킨다. 본 발명의 상황에서, 유전자는 유전자 생성물을 위한 서열뿐만아니라 조절 서열을 포함하는 게놈의 부위로서 정의되고 단백질 또는 RNAs의 생성을 초래한다. 이러한 정의는 당업계에서 일반적 정의에 해당된다.
단백질 형질도입(transduction)도메인-매개된, 인공전사인자(transcription factors)의 새포내 운반은 신규한 패션으로 병리생리학(pathophysiological) 관련 분자를 표적하는 생물의 높은 선택성의 장점을 얻는 새로운 방식이다. ADOA 또는 ADOA 플러스와 같은, OPA1의 단상부족(haploinsufficiency)에 의해 원인이 되는 질병에 대하여, 불충분한 유전자 발현은 상기 장애를 위해 근본적 원인이기 때문에, 현재 접근방법, 예들 들면, 소분자 약물을 사용한 치료가 없을 수 있다. 그러나, 단백질 형질 도입(transduction) 도메인(PTD)의 형태를 표적하는 진보된 약물로 인공전사인자 기술을 쌍을 이루는 것에 의해, OPA1의 단상부족은 인공전사인자를 활성시키고 만약 모든 유전자 복사가 기능적이면 도달될 수 있는 수준으로 남아있는 기능적 유전자 복사의 전사를 증강시켜 분자적 수준에서 직접적으로 말할 수 있다.
고려되는 단백질 형질도입(transduction) 도메인은 HIV TAT, 펩타이드 mT02 (서열번호:18), 펩타이드 mT03 (서열번호:19), R9 펩타이드 (서열번호:20), ANTP 도메인 (서열번호:21) 또는 원형질막을 가로질러 카고(cargo)를 운반(transporting)할 수 있는 다른 펩타이드이다.
추가적으로, 폴리에틸렌 글리콜을 가지는 본 발명의 인공전사인자(transcription factors)의 수정은 면역원성(immunogenicity)을 감소시키는 것으로 고려된다. 추가적으로, 안구 및 뇌와 같은 면역 특권을 가진 기관(immune privileged organs)의 적용은 임의의 면역 반응을 피하고, 인공전사인자(transcription factors)에 대해 전체 인체 내성을 유도할 것이다. 면역 특권을 가진 기관의 밖의 만성 질병 치료에 대하여, 안구내(intraocular) 주사 이전을 통하여 면역 내성의 유도를 고려한다.
우성 시신경 위축(Dominant optic atrophy)은 단상부족으로 유도하는 OPA1유전자에서 돌연변이에 의해 원인이 된다. 우성 시신경 위축 환자는 시신경(optic nerve)을 형성하는 망막 신경절 세포(retinal ganglion cell)의 진행형 결실에 기인하여 맹인에 궁극적으로 원인이 되는 진행형 시력 결실로부터 고통받는다. 흥미롭게도, 대부분의 우성 시신경 위축 환자는 눈-외의(extra-ocular) 증상으로 존재하지 않는다. 환자의 단지 적은 서브세트가 경련성 양측 지 마비(spastic paraplegia) 및 청각 손상과 같은 눈-외의(extra-ocular) 신경학적 증상을 가지는 소위 우성 시신경 위축(optic atrophy) 플러스 형태로부터 고통 받는다. OPA1은 내부 미토콘드리아 크리스태(cristae)의 구조를 안정화하고 미토콘드리아 튜불(tubules)사이의 융합(fusion)을 촉진시켜 구조적 수준에서 미토콘드리아 기능 유지에 관련된다. 미토콘드리아는 ATP 형태에서 세포 에너지의 주요 생성요인이기 때문에, OPA1는 세포의 에너지 수준을 유지하는 것이 필요하다. OPA1 기능의 결실은 세포사멸(apoptotic)기전을 경유하여 세포죽음(cell death)을 촉진하는 것으로 알려져 있다. 인간 몸체의 거의 모든 세포에서 OPA1 유전자의 하나의 기능적 복사(copy)는 충분한 수준에서 미토콘드리아 기능을 유지하는데 충분한 OPA1 단백질을 생성하는데 충분하다. 그러나, 특별히 에너지-부족 망막 신경절(retinal ganglion) 세포는 그들의 미토콘드리아의 상태에 관하여 특수한 필요를 가지고 따라서 하나의 OPA1 유전자 복사에 의해 생성될 수 없는 OPA1의 수준에 의존하고, 따라서 단상부족(haploinsufficiency) OPA1 돌연변이는 망막 신경절(retinal ganglion) 세포죽음(cell death)과 연관되고 시력 결실 및 맹인을 초래한다. 본 발명의 인공전사인자(transcription factors)를 사용하여, OPA1 단백질수준은 남아있는, 정상 수준 이상의 기능적 OPA1 유전자로부터 OPA1 단백질의 생성을 증가시켜 망막 신경절(retinal ganglion)세포에서 증가될 수 있다.
OPA1의 단상부족은 바이러스 이동의 수단에 의한 돌연변이된 OPA1 유전자의 추가의, 기능적 복사 공급을 통하여 전통적 유전자 치료 접근에 의해 이론적으로 치료될 수 있음므로, 따라서 유전자 용량을 증가시킨다. 그러나, 유전자치료를 위하여 안전한 것으로 간주되는 현재 사용가능한 바이러스 벡터는 약 5 내지 8 킬로베이스(kilobases)보다 큰 유전자를 운반할 수 없다. 이것은 일부 유전자에 대하여 충분한 반면에, OPA1 유전자는 8 킬로베이스(kilobases)보다 큰 것으로 고려되고 따라서 현재 사용가능한 벡터를 수용하는 유전자 치료에 대하여 후보가 아니다. 추가적으로, 유전자 발현의 정확한 조절은 운반된 유전자의 과발현의 잠재성 및 연관된 독성 부작용을 가지는 유전자 치료를 사용하여 얻을 수 없다.
바이러스 이동의 상기 제한은 본 발명의 인공전사인자(transcription factors)에 적용하지 않는다. 단상부족 유전자의 크기는 인공전사인자에 의한 조절에 대하여 잘 받아들이는 심지어 가장 큰 유전자를 갖는 본 발명에서 설명된 치료 접근(도 1)에 대하여 관련하지 않는다. 추가적으로, 유전자 발현이 본 발명의 인공전사인자에 의해 증가된 정도는 인공전사인자에 따라서 용량을 통하여 또는 전사 조절 면에서 높거나 낮은 활성을 가지는 대안적(alternative) 활성 도메인을 수용하여 조절된다. 추가적으로, OPA1 mRNA는 자체의 기능을 수행하기 위해서 OPA1에 대하여 모든 필요한 몇 개의 OPA1 아이소폼의 생성의 원인이 되는 대규모 선택적 접합(alternative splicing)으로 가게 한다. 특히, 다양한 OPA1 아이소폼(isoforms)의 차이 있는 단백질 가수 분해의 진행(differential proteolytic processing)은 자체의 기능을 수행하는데 OPA1에 대하여 필수 기계적 전제조건이다.
기능적 유전자 복사에서 OPA1 mRNA 생성 증가에 대하여 본 발명의 인공전사인자의 바이러스 운반을 사용은 발생하는 이러한 필수 과정에 대하여 허용할 것이고, 따라서 OPA1 단상부족에 의해 원인이 되는 질병에 대한 기능적 치유를 제공한다.
치료 제제를 위한 풀(pool)로서 전통적으로 사용된 소분자 분류는 유전자발현의 표적된 조절에 대하여 적절하지 않다. 따라서, 많은 유망한 약물 표적 및 관련 질병은 전형적 약학적 접근에 잘 받아들이지 않는다. 반대로, 본 발명의 인공전사인자는 높게 정의된 전체 조성물을 가지는 동일한 물질 분류에 속한다. 2개의 헥사메릭징크핑거(hexameric zinc finger)단백질-기반인공전사인자(transcription factors)를 표적하는 2개의 매우 다양한 프로모터 서열은 전체 유사한 4차 구조를 가지는 85%의 최소의 아미노산 서열 동일성을 여전히 가지고 빠른 및 경제적 방식에서 표준 방법 (하기에 설명된 바와 같이)을 경유하여 생성될 수 있다. 따라서, 본 발명의 인공전사인자는 분자의 하나의 분류로, 전체 유사한 조성물을 가지는 표적의 매우 넓고 다양한 세트에 대하여 예외적으로 높은 특수성을 조합한다. 추가적으로, 약물로 본 발명의 인공전사인자의 제제는 약물 개발 진행을 추가적으로 촉진하는 이전의 경험에 의존할 수 있다.
또한 본 발명은 폴리닥틸 징크핑거 단백질이 OPA1 프로모터 부위에 특정적으로 표적하기 때문에, OPA1의 단상부족으로 유도하는 OPA1에서 돌연변이에 의해 원인이 되는 질병 치료에서 인공전사인자(transcription factors)의 사용과 관련한다. 이와 같이 본 발명은 본 발명의 인공전사인자의 치료학적으로 유효량을 이를 필요로 하는 환자에게 투여하는 것을 포함하는 질병을 치료하는 방법과 관련하고, 상기 치료되고자 하는 질병은 OPA1 유전자의 단상부족에 의해 원인이 되고, 폴리닥틸 징크핑거 단백질은 OPA1 프로모터에 특정적으로 표적한다.
고려되는 폴리닥틸 징크핑거 단백질은 4분자체(tetrameric), 5분자체(pentameric), 6분자체(hexameric), 7분자체(heptameric) 또는 8분자체(octameric) 징크핑거 단백질이다. "4분자체(tetrameric)", "5분자체(pentameric)", "6분자체(hexameric)", "7분자체(heptameric)" 및 “8분자체(octameric)”는 징크핑거 단백질이 각각 4개, 5개, 6개, 7개 및 8개 부분 단백질 구조로 구성된다는 것을 의미하고, 이것의 각각은 특별한 뉴클레오티드 삼중항(triplet)에 대하여 결합특수성을 가진다. 바람직하게는 인공전사인자는 헥사메릭 징크핑거 단백질을 포함한다.
OPA1 프로모터 부위 내의 표적부위의 선별
표적부위 선별은 기능적 인공전사인자의 성공적 발생을 위하여 결정적이다. 생체내(in vivo) OPA1 유전자 발현을 조절하는 인공전사인자에 대하여, 이것은 OPA1 유전자의 게놈의 상황에서 자체의 표적부위를 결합해야만 한다. 이것은 DNA 표적부위의 접근 가능성을 필요하게 만들고, 이 부위에서 크로모좀의(chromosomal) DNA가 뉴클레오좀(nucleosomes)으로 히스톤(histones) 주위에 단단하게 포장되지 않고 및 인공전사인자결합을 가지는 메틸레이션(methylation) 간섭과 같은 DNA 수정이 없다는 것을 의미한다. 인간게놈의 큰 부분이 단단하게 포장되고 전사적으로 비활성적인 반면, 활발히 전사된 유전자의 전사 시작 부위 (-1000 내지 +200bp)의 즉시 근처는 내인성 전사인자(transcription factors) 및 RNA 폴리머라제와 같은 전사 기계에 대하여 접근 가능함에 틀림없다. 따라서, 임의의 주어진 표적유전자의 상기 영역에서 표적부위를 선별하는 것은 생체내(in vivo) 원하는 기능을 가지는 인공전사인자의 성공적 발생을 허용할 것이다.
인간 OPA1 유전자 프로모터 내의 표적부위 선별
인간 OPA1 개방형 해독틀(open reading frame)의 시작 코돈의 부위 1000 bp 상부(upstream)는 (G/C/ANN)6의 일반 조성물을 가지는 잠재적 18 bp 표적부위의 존재를 위해 분석되었고, 상기 G는 뉴클레오티드 구아닌(guanine), C는 뉴클레오티드 시토신(cytosine), A는 뉴클레오티드 아데닌(adenine) 및 N은 4개의 뉴클레오티드 구아닌(guanine), 시토신(cytosine), 아데닌(adenine) 및 티민(thymine)의 각각을 대표한다. 4개의 표적부위 OPA_TS1 (서열번호:22), OPA_TS2 (서열번호:23), OPA_TS3 (서열번호:24), 및 OPA_TS4 (서열번호:25)가 선택되었다.
OPA1 유전자 프로모터를 표적하는 변환가능한 인공전사인자(artificial transcription factors)
특정한 헥사메릭 징크핑거 단백질은 ZiFit 소프트웨어 v3.3 (Sander JD., Nucleic Acid Research 35, 599-605)를 사용하여 소위 Barbas 징크핑거모듈 세트 (Gonzalez B., 2010, Nat Protoc 5, 791-810)로 구성되었거나 또는 이스트원하이브리드 기술(yeast one hybrid)을 사용하여 징크핑거 단백질 라이브러리로부터 선별되었다. OPA1 유전자 프로모터를 표적하는 변환가능한 인공전사인자를 활성을 생성하기 위하여, 헥사메릭 징크핑거(hexameric zinc finger)단백질 ZFP_OPA1_1 (서열번호:26), ZFP_OPA1_2 (서열번호:27), ZFP_OPA1_3 (서열번호:28), ZFP_OPA1_4 (서열번호:29), ZFP_OPA1_5 (서열번호:30), ZFP_OPA1_6 (서열번호:31), ZFP_OPA1_7 (서열번호:32), ZFP_OPA1_8 (서열번호:33), ZFP_OPA1_9 (서열번호:34), ZFP_OPA1_10 (서열번호:35), ZFP_OPA1_11 (서열번호:36), ZFP_OPA1_12 (서열번호:37), ZFP_OPA1_13 (서열번호:38), ZFP_OPA1_14 (서열번호:39), ZFP_OPA1_15 (서열번호:40), ZFP_OPA1_16 (서열번호:41), ZFP_OPA1_17 (서열번호:42), 및 ZFP_OPA1_18 (서열번호:43),는 인공전사인자 OPA_akt1 (서열번호:44), OPA_akt2 (서열번호:45),OPA_akt3 (서열번호:46),OPA_akt4 (서열번호:47),OPA_akt5 (서열번호:48),OPA_akt6 (서열번호:49),OPA_akt7 (서열번호:50),OPA_akt8 (서열번호:51),OPA_akt9 (서열번호:52),OPA_akt10 (서열번호:53),OPA_akt11 (서열번호:54),OPA_akt12 (서열번호:55),OPA_akt13 (서열번호:56),OPA_akt14 (서열번호:57),OPA_akt15 (서열번호:58),OPA_akt16 (서열번호:59),OPA_akt17 (서열번호:60), 및 또한 NLS 및 3xmyc 에피토프 태그를 포함하는 OPA_akt18 (서열번호:61)을 산출하는 전사 활성 도메인 VP64에 융합된다.
또한 5분자체 또는 6분자체, 7분자체 또는 8분자체 징크핑거단백질을 포함하는 본 발명의 인공전사인자가 고려되고, 상기 각각의 징크핑거 모듈은 OPA1프로모터 유전자의 표적부위에 대하여 결합 친화도를 개선하거나 또는 개선된 내성(tolerability)을 위하여 징크핑거 단백질의 면역학적 프로파일을 변경하는 것으로 교환된다.
또한 본 발명에 따른 OPA1 프로모터를 표적하는 인공전사인자는 서열번호:26 및 43에 개시된 것처럼 징크핑거모듈 조성물에 기반한 징크핑거 단백질을 포함하고, 상기 각각의 아미노산은 의도된 표적부위에 결합 친화도를 고정시키는 동안에 잠재적 면역원성(immunogenicity)을 최소화하기 위하여 교환된다.
또한 본 발명의 인공전사인자는 P16,VP64 (4분자체(tetrameric) repeat of VP16),CJ7, p65-TA1, SAD, NF-1, AP-2, SP1-A, SP1-B, Oct-1, Oct-2, Oct-2_5x, MTF-1, BTEB-2, LKLF. 및 그 외, 바람직하게는 VP64 또는 AP 2와 같은 유전자존재론(ontology) GO:0001071에 의해 정의된 바와 같이 유전자 전사를 증가할 수 있는 다른 단백질 도메인을 포함할 수 있다.
추가적으로, 본 발명의 인공전사인자는 핵국재화서열(nuclear localization sequence) (NLS)를 포함한다. 고려되는 핵국재화서열은 바람직하게는 SV40 NLS와 함께 유전자존재론(ontology) GO:0008139, 예를 들면 리신(lysine) 잔기(K)에 이어서 리신(lysine)(K) 또는 아르기닌(arginine) 잔기 (R), 이어서 임의의 아미노산 (X), 이어서 리신(lysine) 또는 아르기닌(arginine) 잔기 (K-K/R-X-K/R 컨센서스(consensus) 서열, Chelsky D. et al., 1989 Mol Cell Biol 9, 2487-2492) 또는 SV40 NLS (서열번호:62)에 의해 정의된 단백질에 결합을 통하여 핵 수입(nuclear import)을 부여하는 아미노산 모티프이다.
또한 단백질 형질도입(transduction)도메인 없이, PA1 유전자의 프로모터 부위에 직접적인 인공전사인자는 본 발명의 대상(subject)이다. 이들은 윗글에서 정의된 바와 같이, 본 발명의 인공전사인자에 대하여 중간체(intermediates)이고 또는 상기와 같이 사용될 수 있다.
헤르페스 바이러스(herpes virus)-, 아데노 바이러스(adeno virus)- 및 아데노 관련 바이러스(adeno-associated virus)-기반 벡터와 같은, 형질감염 또는 바이러스 벡터를 경유하여 이동된 핵산 형태에서 본 발명의 인공전사인자에 대한 대안적(alternative) 운반방법이 고려된다.
본 발명의 인공전사인자의 도메인은 짧은 유연한 링커(flexible linkers)에 의해 연결될 수 있다. 짧은 유연한 링커는 2 내지 8 아미노산, 바람직하게는 글리신(glycine) 및 세린(serine)을 가진다. 고려되는 특별한 링커는 GGSGGS (서열번호:63)이다. 인공전사인자는 이들의 검출 및 진행을 용이하도록 마커를 추가적으로 포함할 수 있다.
OPA1 프로모터를 표적하는 인공전사인자를 가지는 하기의 치료를 OPA1 상향조절 및 개선된 미토콘드리아 활성 평가
OPA1 프로모터 특정 인공전사인자로 처리된 HeLa 세포는 버퍼 대조군 처리된 세포와 비교될 것이고 OPA1의 단백질 수준은 특정한 항-OPA1 항체를 사용하여 정량적 적외선(infrared)-형광 기반된 웨스턴 블랏에 의해 평가될 것이다. OPA1 단백질 수준에서 증가는 OPA1의 생성 증가에 이어서 인공전사인자로 치료의 표시(indicative)이다. OPA1 특정한 인공전사인자를 가지는 치료의 이익적 효과를 측정하기 위하여, 미토콘드리아 정확도 및 세포의 생존은 평가된다. 이를 위하여, OPA1 특정 인공전사인자로 처리된 세포는 미토콘드리아 독성 로테논(poison rotenone)을 가지는 치료를 통하여 계기가 되는 미토콘드리아 활성 산소(reactive oxygen) 생성에 이은 산화 공격(oxidative insult)의 면에서 대조군 처리된 세포와 비교되었다. 미토콘드리아 활성 산소 생성은 유동 분석(flow cytometry) 및 활성 산소 특정 염색제 MitoSox를 사용하여 측정되었다. 추가적으로, 미토콘드리아 건강의 파라미터(parameter)로서 미토콘드리아 막 전위(mitochondrial membrane potential)는 전위-민감 TMRE 형광의 유동 세포 측정 검출에 의해 측정되었다. 대조군 세포와 비교하여 인공전사인자 처리된 세포에서 활성 산소(reactive oxygen) 종 생성의 감소 또는 미토콘드리아 막 전위(mitochondrial membrane potential)의 증가는 OPA1-표적 인공전사인자의 이익적 활성의 표시이다. 추가적으로, OPA1-표적 인공전사인자 또는 대조군 처리된 세포의 스타우로스포린(staurosporine), 로테논(rotenone) 및 악티노마이신 D(actinomycin D)에 의한 세포사멸(apoptotic) 유도에 대한 민감성은 측정된다. 이를 위하여, 세포사멸의 지표로서 사이토크롬 c(cytochrome c)의 배출은 처리된 세포의 형광 현미경을 사용하여 측정되었고 대조군 세포와 비교되었다.
폴리에틸렌 글리콜 잔기의 부착
본 발명의 인공전사인자에 폴리에틸렌 글리콜잔기(PEGylation)의 공유결합 부착은 인공전사인자의 용해도를 증가시키고, 자체의 신장 청정(renal clearance)을 감소시키고, 자체의 면역원성(immunogenicity)을 조절하는 것으로 고려된다. 1부터 40 킬로달톤(Kilodalton)까지의 크기 범위의 티올(thiol) 반응 폴리에틸렌 글리콜(polyethylene glycols)뿐만아니라 아민(amine)이 고려된다. 티올(thiol) 반응 폴리에틸렌 글리콜을 사용하여, 인공전사인자의 부위-특정 PEGylation를 얻는다. 본 발명의 인공전사인자에서 아미노산을 포함하는 단지 필수 티올기(thiol group)는 아연 조화(zinc coordination)에 대하여 필수적인 징크핑거 모듈에 위치된 시스테인 잔기이다. 상기 티올기(thiol group)는 아연 조화(zinc coordination)에 기인하여 PEGylation에 대하여 접근 가능하지 않으므로, 따라서 본 발명의 인공전사인자로 하나 또는 몇개의 시스테인 잔기의 포함은 티올(thiol)-특정 폴리에틸렌 글리콜 시약을 사용하여 PEGylation을 위하여 티올기가 없는 것을 제공한다.
약학적 조성물
본 발명은 상기에서 정의된 바와 같이 인공전사인자를 포함하는 약학적 조성물에 관한 것이다. 고려되는 약학적 조성물은 온혈동물, 특히 인간에게 비경구 시스테믹(parenteral systemic) 투여, 특히 정맥내(intravenous) 투여, 흡입을 위한 조성물, 및 국소 투여를 위한 조성물, 특히 국소용 안과용(ophthalmic-topical) 투여, 예들 들면, 안약(eye drops), 또는 유리체내(intravitreal), 결막밑의(subconjunctival), 안구옆쪽의(parabulbar) 또는 안구뒤쪽의(retrobulbar) 투여를 위한 조성물이다. 특별히 바람직한 것은 유리체내(intravitreal), 결막밑의(subconjunctival), 안구옆쪽의(parabulbar) 또는 안구뒤쪽의(retrobulbar) 투여를 위한 안약(eye drops) 및 조성물이다. 조성물은 단독 또는, 바람직하게는, 약학적으로 허용가능한 담체와 같이 유효 성분을 포함한다. 추가적으로 느린-배출 제제(formulations)가 고려된다. 유효 성분의 용량은 치료되고자 하는 질병 및 종, 자체의 연령, 체중, 및 각각의 조건, 각각의 약동학 데이타, 및 투여 모드에 의존한다.
추가적으로 경구 운반(oral delivery)에 유용한 약학적 조성물, 장에서 적합하게 캡슐에 넣어진 유효성분, 또는 그렇지 않으면 분해에 대하여 보호되는 것을 포함하는 조성물이 고려된다. 예를 들면, 상기 약학적 조성물은 막 투과성 증강시키는 제제, 프로테아제 효소 억제제를 포함할 수 있고, 및 장용성(enteric)코팅에 의해 감싸지게 된다(enveloped).
약학적 조성물은 대략 1%부터 대략 95%까지의 유효성분을 포함한다. 단위용량(Unit dose) 형태는, 예를 들면, 앰폴(ampoules), 바이알(vials), 흡입기(inhalers), 안약(eye drops) 등이다.
본 발명의 약학적 조성물은 자체로 알려진 방식, 예를 들면 종래의 혼합, 용해 또는 동결건조 과정에 의해 제조된다.
선호되는 것은 유효성분의 용액의 사용에 주어지고, 및 또한 현탁액 또는 분산제(dispersions), 특히 수분을 함유한 등장성(isotonic aqueous)용액, 분산제(dispersions) 또는 현탁액, 예를 들면 유효성분 단독 또는 담체와 함께, 예를 들면 만니톨(mannitol)의 사용은 사용 전에 만들어 질 수 있다. 약학적 조성물은 살균될 수 있고 및/또는 부형제(excipients), 예를 들면 보존제(preservatives), 안정제(stabilizers), 습윤(wetting) 제제 및/또는 유화제(emulsifiers), 용해제(solubilizers), 삼투압(osmotic pressure) 조절을 위한 염 및/또는 버퍼를 포함할 수 있고 알려진 방식 자체로, 예를 들면 종래의 용해 및 동결건조 과정의 수단에 의해 제조된다. 상기 용액 또는 현탁액은 점도(viscosity)-증가하는 제제, 통상적으로 소듐 카르복시메틸세룰로오스(sodium carboxymethylcellulose), 카르복시메틸세룰로오스(carboxymethylcellulose), 덱스트란(dextran), 폴리비닐피롤뢰딘(polyvinylpyrrolidone), 또는 젤라틴(gelatins), 또는 또한 용해제(solubilizers), 예들 들면, 트윈 80™(polyoxyethylene(20)sorbitan 단일(mono)-oleate)를 포함할 수 있다.
오일에 있는 현탁액은 오일 구성요소로서 주사 목적을 위하여 채소, 합성, 또는 반-합성 오일 관습을 포함한다. 상기 관점에서, 특수한 언급은 산 구성요소로서 8부터 22까지, 특히 12부터 22까지, 탄소 원자를 가지는 긴-사슬지방산을 포함하는 액체 지방산 에스테르로 만들어질 수 있다. 상기 지방산 에스테르의 알코올 구성요소는 최대 6 탄소 원자를 가지고 1가의 또는 다가(polyvalent), 예를 들면 단일(mono)-, 2중-(di-) 또는 3중(trivalent), 알코올, 특히 글리콜 및 글리세롤이다. 지방산 에스테르의 혼합물로서, 목화씨 오일, 아몬드 오일, 올리브 오일, 피마자유, 참기름, 대두유 및 땅콩오일과 같은 채소 오일은 특히 유용하다.
주사용 제제의 제작은 예를 들면, 앰폴(ampoules) 또는 바이알(vials)로 충전, 및 용기의 밀봉과 같은, 멸균 조건하에서 항상 수행된다.
비경구 투여를 위하여, 점도(viscosity)-증가 물질, 예를 들면 소듐 카르복시메틸세룰로오스(sodium carboxymethylcellulose), 소르비톨(sorbitol) 및/또는 덱스트란(dextran), 및 만약 원한다면 안정제(stabilizers)를 포함하는 수용성 형태(water-soluble form), 예를 들면 수용성염(water-soluble salt), 또는 수용성 주사(aqueous injection) 현탁액에 있는 유효 성분의 수용액은 특히 적절하다. 또한 선택적으로 부형제와 함께, 유효성분은 동결건조(lyophilizate)의 형태일 수 있고 적절한 용매의 첨가에 의해 비경구 투여 전에 용액으로 만들어질 수 있다.
흡입을 위한 조성물은 에어로졸 형태, 스프레이(sprays). 미스트(mist) 또는 드롭(drops) 형태로 투여될 수 있다. 에어로졸은 계량기된-용량 흡입기(metered-dose inhaler) 또는 분무기(nebulizer), 즉, 적절한 추진체(propellant), 예들 들면, 환자에 의해 흡입된 에어로졸된 의약의 짧은 분출의 형태로, 디클로로디플루오로-메탄(dichlorodifluoro-methane), 트리클로로플루오로메탄(trichlorofluoromethane), 디클로로테트라플루오로에탄(dichlorotetrafluoroethane), 이산화탄소(carbon dioxide) 또는 다른 적절한 가스를 사용하여 기도(airways) 또는 폐(lungs)에 의약의 특정한 양을 운반하는 장치로 운반될 수 있는 용액 또는 현탁액으로부터 제조된다. 또한 이것은 락토오스(lactose) 또는 전분(starch)과 같은 적절한 분말로 흡입을 위한 부말 스프레이를 제공하는 것이 가능하다.
바람직하게는 안약(eye drops)은 라크리말 유동액(lacrimal fluid)(295-305 mOsm/l)을 가지는 조성물 등장액을 제공하는 적절한 제제를 포함하는 유효 성분의 등장성(isotonic aqueous)용액을 함유한 수분이다. 고려되는 제제는 소듐 클로라이드(sodium chloride), 시트르산(citric acid), 글리세롤, 소르비톨(sorbitol), 만니톨(manitol), 에틸렌 글리콜, 프로필렌 글리콜, 덱스트로오스 등이다. 추가적으로 조성물은 5와 8 사이, 바람직하게는 7.0 내지 7.4의 pH를 유지하기 위하여 버퍼링 제제, 예를 들면 포스페이트 버퍼, 포스페이트-시트레이트, 또는 트리스 버퍼 (tris(hydroxymethyl)-aminomethane)를 포함한다. 조성물은 항미생물 보존제(preservatives), 예를 들면 파라벤(parabens), 벤잘코니윰 클로라이드(benzalkonium chloride), 폴리헥사메틸린 비구아닌(polyhexamethylene biguanidine (PHMB)) 등과 같은 4차 암모늄(quaternary ammonium) 염을 추가적으로 포함할 수 있다. 안약(eye drops)은 젤-같은 안약, 및/또는 히아루론산(hyaluronic acid), 메틸셀룰로오스(methylcellulose), 폴리비닐알코올(polyvinylalcohol), 또는 폴리비닐피롤리딘(polyvinylpyrrolidone)과 같은 다른 점도(viscosity) 증강시키는 제제를 생성하기 위하여 쟌탄 검(xanthan gum)을 추가적으로 포함할 수 있다.
치료 방법에서 인공전사인자(transcription factors)의 사용
추가적으로 본 발명은 OPA1 생성의 증가를 위한 사용, OPA1에 의해 영향받는 질병의 치료에 사용, 특히 안구 질병의 치료에 사용을 위하여 상기에서 설명된 것처럼 OPA1 프로모터에 직접적으로 인공전사인자(transcription factors)에 관련된다. OPA1에 의해 조절되는 질병은 정상 긴장상태 녹내장(normal tension glaucoma)뿐만아니라, 상염색체성우성시신경위축(autosomal dominant optic atrophy), 상염색체성우성시신경위축(autosomal dominant optic atrophy plus) 플러스이다.
이와 같이 본 발명은 이를 필요로 하는 환자에게 본 발명의 인공전사인자의 치료학적으로 유효량을 투여하는 것을 포함하는 OPA1에 의해 영향받는 질병을 치료하는 방법과 관련된다. 특히 본 발명은 정상 긴장상태(tension) 녹내장(normal tension glaucoma) 또는 우성시신경위축으로 연관된 신경퇴화를 치료하는 방법과 관련된다. 본 발명의 인공전사인자의 유효량은 치료되고자 하는 질병의 특정 유형 및 종, 자체의 연령, 체중, 및 각각의 조건, 각각의 약동학 데이타, 및 투여 모드 에 의존된다. 안구로 투여를 위하여, 0.5 내지 1 mg의 매월 유리체 주사(vitreous injection)가 바람직하다. 시스테믹 적용을 위하여, 10 mg/kg의 매월 주사가 바람직하다. 추가적으로, 안구의 유리체(vitreous)로 느린 배출 침착물의 이식이 또한 바람직하다.
동물에서 인공전사인자의 사용
추가적으로 본 발명은 유전자 생성물 발생을 증가시키기 위하여, 동물 OPA1프로모터를 표적하는 인공전사인자의 사용과 관련된다. 바람직하게는, 인공전사인자는 이를 필요로 하는 동물에게 국소적용을 위하여 절한 조성물에 직접적으로 적용된다.
실시예
DNA 플라스미드의 클로닝
모든 클로닝 단계를 위하여, 제한 엔도뉴클레아제(restriction endonucleases) 및 T4 DNA 결찰효소(T4 DNA ligase)를 New Engl및 Biolabs으로부터 구입하였다. 스림프 알칼린포스파타제(Shrimp Alkaline Phosphatase(SAP))는 Promega로부터이다. 높은-정확도 플라티늄 Pfx DNA 폴리머라제(Invitrogen)은 모든 표준 PCR 반응에서 적용된다. DNA 단편 및 플라스미드는 뉴클레오스핀 젤(NucleoSpin Gel) 및 PCR 클린업 키트(PCR Clean-up kit), 뉴클레오스핀플라스미드 키트(NucleoSpin Plasmid kit), 또는 뉴클레오본드엑스트라 미디플러스 키트(NucleoBond Xtra Midi Plus kit) (Macherey-Nagel)를 사용하여 제작사의 지침에 따라 분리된다. 올리고뉴클레오티드를 Sigma-Aldrich로부터 구입하였다. 새롭게 생성된 플라스미드의 모든 관련 DNA 서열은 시퀀싱에 의해 증명되었다(Microsynth).
이스트원하이브리드 (yeast one hybrid)를 위하여 헥사메릭 징크핑거 단백질(hexameric zinc finger protein)의 클로닝
GNN및/또는 CNN 및/또는 ANN 결합징크핑거(ZF) 모듈을 포함하는 헥사메릭 징크핑거 단백질 라이브러리는 하기의 개선으로 Gonzalez B. et al,.2010, Nat Protoc 5, 791-810따른 클론이다. GNN, CNN 및 ANN ZF 모듈을 위하여 DNA 서열 코딩을 합성하였고 pUC57 (GenScript)로 삽입하였고 각각 pAN1049 (서열번호:64), pAN1073 (서열번호:65) 및 pAN1670 (서열번호:66)을 초래하였다. 징크핑거단백질 (ZFP)라이브러리의 단계적 어셈블리는 pBluescript SK (+) 벡터에서 행하여졌다. 비-기능적 단백질을 유도하는 각각의 클로닝 단계동안에 다수의 ZF 모듈의 삽입을 피하기 위하여, pBluescript (및 1ZFP, 2ZFPs, 또는 3ZFP를 포함하는 자체의 유도된 생성물) 및 pAN1049, pAN1073 또는 pAN1670는 하나의 제한 효소 및 이후에 처리된 SAP으로 첫 번째 배양되었다. 효소는 두번째 제한 엔도뉴클레아제(restriction endonucleases)를 첨가하기 전에 뉴클레오스핀 젤(NucleoSpin Gel) 및 PCR 클린업 키트(PCR Clean-up kit)를 사용하여 제거하였다.
pBluescript-1ZFPL의 클로닝은 5 ㎍ pBluescript를 XhoI, SAP 및 그 뒤에 SpeI로 처리하여 수행된다. 삽입은 10 ㎍ pAN1049 (16개의 상이한 GNN ZF 모듈의 배출) 또는 pAN1073 (15개의 상이한 CNN ZF 모듈의 배출) 또는 pAN1670 (15개의 상이한 ANN ZF 모듈의 배출)을 SpeI, SAP 및 그 뒤에 XhoI으로 배양하여 생성된다. pBluescript-2ZFPL 및 pBluescript-3ZFPL의 생성을 위하여, 7 ㎍ pBluescript-1ZFPL 또는 pBluescript-2ZFPL을 AgeI으로 절단되고, 탈인산화(dephosphorylated)되고, SpeI으로 절단된다. 삽입은 각각 10 ㎍ pAN1049 또는 pAN1073 또는 pAN1670으로 SpeI, SAP, 및 그 뒤에 XmaI을 적용하여 얻는다. pBluescript-6ZFPL의 클로닝은 잘려진 벡터를 얻기 위하여 14 ㎍의 pBluescript-3ZFPL를 AgeI, SAP, 및 이후에 SpeI으로 처리하여 수행된다. 3ZFPL 삽입은 SpeI, SAP, 및 그 뒤에 XmaI로 배양하여 20 ㎍의 pBluescript-3ZFPL로부터 배출되었다.
하나, 2개, 및 3개 ZFPs를 포함하는 라이브러리를 위한 결찰(ligation) 반응은 밤사이 상온에서 20 ㎕ 총 부피에서 200 ng 잘려진 벡터, 400 U T4 DNA 결찰효소를 사용하여 삽입체:벡터의 3:1 몰라 비율로 셋업하였다. 헥사메릭징크핑거단백질 라이브러리의 결찰(ligation) 반응은 200 ㎕ 총 부피에서 2000 ng pBluescript-3ZFPL, 500 ng 3ZFPL 삽입체, 4000 U T4 DNA 결찰효소를 포함하고, 이것을 20 ㎕씩 10회 나누고 밤사이 상온에서 별도로 배양하였다. 결찰(ligation) 반응의 부분은 각각의 라이브러리를 위하여 요구되는 클론 수에 의존하여 몇 가지 방법에 의해 박테리아(Escherichia coli)로 형질전환되었다. pBluescript-1ZFPL 및 pBluescript-2ZFPL의 발생을 위하여, 3 ㎕의 결찰(ligation) 반응을 E. coli NEB 5-알파의 열 충격(heat shock) 형질전환을 위해 직접적으로 사용하였다. pBluescript-3ZFPL의 결찰(ligation) 반응의 플라스미드 DNA는 뉴클레오스핀 젤(NucleoSpin Gel) 및 PCR 클린업 키트(PCR Clean-up kit)를 사용하여 정제되었고 전기컴피턴트(electrocompetent) E. coli NEB 5-알파 (EquiBio로부터 EasyjecT 플러스 전기천공법(electroporator) 또는 Eppendorf로부터 Multiporator, Bio-Rad로부터 2.5 kV 및 25 μF, 2 mm 전기천공법 큐벳(electroporation cuvettes))로 형질전환되었다. pBluescript-6ZFP 라이브러리의 결찰 반응은 뉴클레오스핀 젤(NucleoSpin Gel) 및 PCR 클린업 키트(PCR Clean-up kit)으로 적용되었고 DNA는 15㎕의 탈이온수(deionized water)로 용출되었다. 약 60 ng의 염분을 제거한 DNA는 50 ㎕ NEB 10-베타 전기컴피턴트(electrocompetent) E. coli (New Engl및 Biolabs)으로 혼합되고 전기천공은 EasyjecT 플러스 또는 Multiporator, 2.5 kV, 25 μF 및 2 mm ㅈ전기천공 큐벳(ectroporation cuvettes)을 사용하여 제작사에 의해 추천되는 바와 같이 수행된다. 다수의 전기천공(electroporations)은 각각 라이브러리에 대하여 수행되었고 세포는 직접적으로 풀(pool)되었고 그 후에 라이브러리 크기를 증가되었다. 열 충격 형질전환 또는 전기천공 후에 SOC 배지는 박테리아에 적용되었고 37℃ 및 250 rpm에서 1 시간 배양 후에, 30 ㎕의 SOC 배양은 연속적 희석에 사용되었고 앰피실린(ampicillin)을 포함하는 LB 플레이트에 플레이트하였다. 다음날, 얻은 라이브러리 클론의 총수를 결정하였다. 추가적으로, 각각의 라이브러리의 10개의 클론을 선택하여 플라스미드 DNA를 분리하고 제한 효소 분해로 삽입체의 혼합을 체크하였다. 적어도 상기 플라스미드의 3개를 라이브러리의 다양성을 증명하기 위하여 시퀀싱하였다. 남아있는 SOC 배양을 앰피실린이 포함된 100 ml LB 배지로 이동시켰고 37℃ 및 250 rpm에서 밤사이 동안 배양하였다. 상기 세포를 각각의 라이브러리에 대하여 플라스미드 Midi DNA를 제조하는데 사용하였다.
이스트원하이브리드 스크리닝을 위하여, 헥사메릭 징크핑거 단백질은 프레이 벡터(prey vector)로 호환이 되도록 이동된다. 상기 목적을 위하여 pGAD10 (Clontech)의 다수의 클로닝 부위는 XhoI/EcoRI로 벡터를 절단하고 애닐된(annealed) 올리고뉴클레오티드s OAN971 (TCGACAGGCCCAGGCGGCCCTCGAGGATATCATGATG ACTAGTGGCCAGGCCGGCCC, 서열번호:67) 및 OAN972 (AATTGGGCCGGC CTGGCCACTAGTCATCATGATATCCTCGAGGGCCGCCTGGGCCTG, 서열번:68)로 삽입하여 수정되었다. 얻어진 백터 pAN1025 (서열번호:69)는 탈인산화되고 6ZFP 라이브러리 삽입체는 XhoI/SpeI에 의해 pBluescript-6ZFPL로부터 배출되었다. NEB 10-베타 전기컴피턴트(electrocompetent) E. coli로 결찰(ligation) 반응 및 전기천공(electroporations)은 pBluescript-6ZFP 라이브러리에 대하여 상기 설명된 것처럼 수행되었다.
개선된 이스트원하이브리드 스크리닝을 위하여, 헥사메릭 징크핑거(hexameric zinc finger)라이브러리는 개선된 프레이 벡터(prey vector) pAN1375 (서열번호:70)으로 또한 이동된다. 상기 프레이 벡터(prey vector)는 하기와 같이 구성되었다: pRS315 (서열번호:71)은 ApaI/NarI으로 절단되고 애닐된(annealed) OAN1143 (CGCCGCATGCATTCATGCAGGCC, 서열번호:72) 및 OAN1144 (TGCATGAATGCATGCGG, 서열번호:73)는 삽입되어 pAN1373 (서열번호:74)를 산출하였다. pAN1025으로부터 SphI 삽입체는 SphI으로 잘려진 pAN1373 절단으로 결찰되었고 pAN1375을 얻었다.
추가적으로 개선된 이스트원하이브리드 스크리닝을 위하여, 헥사메릭징크핑거(hexameric zinc finger)라이브러리는 개선된 프레이 벡터 pAN1920 (서열번호:75)로 또한 이동된다.
더욱 추가적으로 개선된 이스트원하이브리드 스크리닝을 위하여, 헥사메릭 징크핑거 라이브러리는 프레이 벡터 pAN1992 (서열번호:76)으로 삽입되었다.
이스트원하이브리드 스크리닝을 위하여 배이트 플라스미드(bait plasmids )의 클로닝
각각의 배이트 플라스미드(bait plasmids)를 위하여, 중심에 있는 18 bp의 잠재성인공전사인자표적부위를 포함하는 60 bp 서열이 선별되고 NcoI 부위는 제한 분석을 위하여 포함된다. 올리고뉴클레오티드는 HindIII/XhoI으로 절단된 pAbAi (Clontech)으로 직접적 결찰(ligation)을 허용하는 5’ HindIII 및 3’ XhoI 부위를 생성하기 위하여 상기 방식으로 디자인되었고 애닐되었다(annealed). NcoI를 가지는 생성물의 분해 및 시퀀싱은 배이트 플라스미드(bait plasmids)의 어셈블리를 확인하기 위하여 사용되었다.
효모 계통(strain) 및 배지
Saccharomyces cerevisiae Y1H Gold를 Clontech로부터 구입하였고, YPD 배지 및 YPD 아가를 CarlRoth로부터 구입하였다. 합성 드롭-아웃(drop-out) (SD) 배지는 20 g/l 글루코오스, 6.8 g/l Na2HPO4 -2H2O, 9.7 g/l NaH2PO4 -2H2O (Carl Roth로부터 모두), 1.4 g/l 효모 합성 드롭-아웃(drop-out) 배지 보충제, 6.7 g/l 효모 질소 베이스(nitrogen base), 0.1 g/l L 트립토판(tryptophan), 0.1 g/l L-루신, 0.05 g/l L-아데닌(adenine), 0.05 g/l L-히스티딘, 0.05 g/l 우라실(Sigma-Aldrich로부터 모두)를 포함하였다. SD U 배지는 우라실(uracil)을 제외하고 모든 구성요소를 포함하였고, SD-L은 L-루신없이 제조되었다. SD 아가 플레이트는 소듐 포스페이트를 포함하지 않았으나, 16 g/l 박토 아가(Bacto Agar (BD))를 포함하였다. 아우레오바시딘 A(Aureobasidin A (AbA))는 Clontech로부터 구입되었다.
배이트 (bait) 효모 계통의 제조
각각 배이트 플라스미드(bait plasmid)의 약 5 ㎍은 20 ㎕의 총 부피에서 BstBI으로 직선화하였고 반응 혼합물의 반은 S. cerevisiae Y1H Gold의 열 충격 형질전환을 위하여 직접적 사용되었다. 효모세포는 변형 하루 전에 5 ml YPD 배지를 접종하는데 사용하였고 상온에서 롤러(roller) 상에서 밤사이 자라게 하였다. 상기 전배양(pre-culture)의 1 밀리리터(milliliter)는 신선한 YPD 배지로 1:20으로 희석되었고 2-3 시간 동안 30℃, 225 rpm에서 배양되었다. 각각 변형 반응을 위하여 1 OD600세포는 원심분리로 수확되었고, 효모세포는 1 ml 멸균수로 1회, 1 ml TE/LiAc (10 mM Tris/HCl, pH 7.5, 1 mM EDTA, 100 mM 리튬 아세테이트)로 1회 세척되었다. 최종적으로, 효모세포는 50 ㎕ TE/LiAc로 재현탁되었고 salmon testes (Sigma-Aldrich)로부터 50 ㎍ 단일가닥 DNA, 10 ㎕의 BstBI-직선화된 배이트 플라스미드 (상기를 참조), 및 300 ㎕ PEG/TE/LiAc(10 mM Tris/HCl, pH 7.5, 1 mM EDTA, 100 mM 리튬 아세테이트, 50% (w/v) PEG 3350)로 혼합되었다. 세포 및 DNA를 상온에서 20 분 동안 롤러(roller) 상에서 배양하였고, 그 이후에 15 분동안 42℃ 항온 수조(water bath)로 옮겼다. 최종적으로, 효모세포를 원심분리로 수득하였고, 100 ㎕ 멸균수에서 재현탁하였고 SD-U 아가 플레이트 상에서 펄쳤다. 30℃에서 배양 3일 후에 각각의 변형 반응으로부터 SD-U에서 자라는 8개 클론을 선택하여 아우레오바시딘 A(aureobasidin A (AbA))에 대한 자체의 민감성을 분석하도록 선택되었다. 전-배양( Pre-cultures)은 상온에서 롤러 상에서 밤사이 자랐다. 각각의 배양동안에, OD600을 측정하였고 OD600=0.3을 멸균수로 조정하였다. 상기 첫번째 희석으로부터 5번 추가 1/10 희석 단계를 멸균수로 제조하였다. 각각 클론에 대하여 각각 희석 단계로부터 SD U, SD U 100 ng/ml AbA, SD U 150 ng/ml AbA, 및 SD U 200 ng/ml AbA를 포함하는 아가 플레이트 상에서 스팟하였다. 30℃에서 3일 동안 배양한 후에, SD-U 상의 웰에서 자라고 및 AbA에 가장 민감한 3개의 클론은 추가 분석을 위하여 선택되었다. 효모게놈의 배이트 플라스미드(bait plasmids)의 안정적 통합(Stable 통합)은 제작사의 지침에 따른 Matchmaker Insert Check PCR Mix 1(Clontech)에 의해 증명되었다. 3개의 클론의 하나를 후속적 Y1H 스크린에 대하여 사용하였다.
헥사메릭징크핑거단백질을 가지는 배이트 효모 계통(bait yeast strain)의 변형
효모 배이트 계통 전배양(bait strain pre-culture)의 약 500 ㎕는 1 l YPD 배지로 희석되었고 OD600=1.6-2.0 (circa 20 시간)까지 30℃ 및 225 rpm에서 배양되었다. 세포를 스윙-아웃 로터(swing-out rotor) (5 분, 1500?g, 4℃)에서 원심분리하여 수득하였다. 전기컴피턴트 세포의 제조는 Benatuil L. et al., 2010, 단백질 Eng Des Sel 23, 155-159에 따라 수행되었다. 각각 변형 반응에 대하여, 400 ㎕ 전기컴피턴트(electrocompetent) 배이트 효모세포를 6ZFP 라이브러리를 암호화하는 1 ㎍ 프레이 플라스미드로 혼합되었고 3 분 동안 얼음에서 배양하였다. 세포-DNA 현탁액을 미리-차가위진 2 mm 전기천공 큐벳(electroporation cuvette)으로 이동시켰다. 다수의 전기천공 반응(EasyjecT 플러스 전기천공기(electroporator) 또는 멀티포레이터(Multiporator), 2.5 kV 및 25 μF)은 모든 효모세포 현탁액이 형질전환되가까지 수행되었다. 전기천공 후에 효모세포를 YPD:1 M 소르비톨의 1:1 혼합의 100 ml로 이동시켰고 60 분 동안 30℃ 및 225 rpm에서 배양하였다. 세포를 원심분리하여 수즉하였고 SD-L 배지의 1-2 ml에서 재현탁하였다. 200 ㎕의 앨리쿼트를 1000-4000 ng/ml AbA를 포함하는 15 cm SD-L 아가 플레이트에 펄쳤다(spread). 추가적으로, 세포 현탁액의 50 ㎕를 1/100 및 1/1000 희석을 만드는데 사용되었고 비희석되고 및 희석된 세포의 50 ㎕를 SD-L에 플레이트하였다. 모든 플레이트를 3일 동안 30℃에서 배양하였다. 얻어진 클론의 총 수는 희석된 변형체(transformants)가 있는 플레이트로부터 계산하였다. 비희석된 세포를 가지는 SD-L 플레이트는 모든 변형체(transformants)의 성장을 나타내는 반면에, 만약 프레이 6ZFP가 자체의 배이트 표적부위에 성공적으로 결합하지 않는다면 단지 AbA-포함하는 SD-L 플레이트는 콜로니 형성을 초래하였다.
6ZFP -암호화하는 프레이 (prey) 플라스미드의 양성 상호작용 및 회수의 증명
초기 분석을 위하여, 40개 좋은 크기의 콜로니를 가장 높은 AbA 농도를 포함하는 SD-L 플레이트로부터 집어서(pick) 효모세포를 1000-4000 ng/ml AbA를 가지는 SD-L 상에 2회 재스트리킹하여(restreaked) 단일 콜로니를 얻었다. 각각 클론을 위하여, 하나의 콜로니는 5 ml SD-L 배지에 접종하는데 사용되었고 세포를 밤사이 상온에서 자라게 하였다. 다음날, OD600=0.3을 멸균수로 조정되었고, 5회 추가 1/10 희석이 제조되었고 각각의 희석 단계의 5 ㎕는 SD-L, SD-L 500 ng/ml AbA, 1000 ng/ml AbA, SD-L 1500 ng/ml AbA, SD-L 2000 ng/ml AbA, SD-L 2500 ng/ml AbA, SD-L 3000 ng/ml AbA, 및 SD-L 4000 ng/ml AbA 플레이트에 스팟되었다(spotted). 클론을 높은 AbA 농도 상에서 자라는 그들의 능력에 따라 순위를 매겼다. 가장 잘 자라는 클론으로부터 초기 SD-L 전배양(pre-culture)의 5 ml는 세포를 스핀 다운하고 100 ㎕ 물 또는 잔여 배지에 이들을 재현탁하는데 사용되었다. 50 U 리티케이스(lyticase) (Sigma-Aldrich, L2524)를 첨가 후에 세포를 수평방향의 쉐이커(horizontal shaker) 상에서 37℃ 및 300 rpm에서 몇시간 동안 배양하였다. 발생된 스페로플라스트(spheroblasts)는 10 ㎕ 20% (w/v) SDS 용액을 첨가하고, 1분 동안 볼텍스하여 격렬하게 혼합하고 적어도 1시간 동안 -20℃에서 냉동시켰다. 그 이후에, NucleoSpin 플라스미드 키트로부터 250 ㎕ A1 버퍼 및 유리 비드(glass beads)의 하나의 스파튤라 팁(patula tip) (Sigma-Aldrich, G8772)을 첨가하였고 튜브를 1 분 동안 볼텍싱하여 격렬하게 혼합하였다. 플라스미드 분리는 표준 뉴클레오스핀 플라스미드 키트 프로토콜로 계속하기 전에 뉴클레오스핀 플라스미드 키트로부터 250 ㎕ A2 버퍼를 첨가하고 상온에서 적어도 15 분 동안 배양하여 추가적으로 개선된다. 용리 버퍼 5 ㎕의 30 ㎕로 용리한 후에 플라스미드 DNA를 열 충격(heat shock) 형질전환에 의해 E. coli DH5 알파로 형질전환된다. 2개의 각각의 콜로니는 앰피실린(ampicillin)-포함하는 LB로부터 피크(pick)되고, 플라스미드는 분리되고 라이브러리 삽입체를 시퀀싱하였다. 얻어진 결과를 각각 표적부위에 대하여 6ZFPs 중에서 컨센서스(consensus) 서열에 대하여 분석하였다.
혼합된 배출된 루시퍼라제 및 알칼린포스파타제 분석을 위한 OPA1 유전자프로모터 부위의 클로닝
OPA1 프로모터 부위를 포함하는 DNA 단편을 pAN1485(NEG-PG04, 유전자Copeia)로 클로닝하였고 이것은 PA1 유전자 프로모터의 대조군 하에서 배출된 가우시아 (Gaussia) 루시퍼라제 및 구성하는 CMV 프로모터의 대조군하에서 배출된 배아 알칼린 포스파타제를 포함하는 리포터플라스미드 pAN1680 (서열번호:77)를 초래하고 이것은 알칼린 포스파타제 신호에 대하여 루시퍼라제의 정상화를 허용한다.
포유류 형질감염을 위한 인공전사인자(transcription factors)의 클로닝
Gen합성 (GenScript)을 통하여 생성되거나 이스트원하이브리드에 의해 선별되는 폴리닥틸 징크핑거 단백질을 암호화하는 DNA 단편은 관심있는 징크핑거 어레이, SV40 NLS, 3x myc 에피토프 태그와 N-말단 KRAB 도메인 (pAN1255 - 서열번호:78), C-말단 KRAB 도메인 (pAN1258 - 서열번호:79), SID 도메인 (pAN1257 - 서열번호:80) 또는 VP64 활성 도메인 (pAN1510 - 서열번호:81) 사이에 융합(fusion)단백질로서 포유류 세포에서 발현을 위하여 포유류 발현벡터로 AgeI/XhoI로 표준 절차를 사용하여 클론되었다.
안정적 형질감염의 발생을 위하여 플라스미드, 테트라사이클린-유도 세포를 하기와 같이 생성하였다: 폴리닥틸 징크핑거 도메인, 조절 도메인 (N-말단 KRAB, C-말단 KRAB, SID 또는 VP64), SV40 NLS 및 3x myc 에피토프 태그를 포함하는 인공전사 인자를 암호화하는 DNA 단편은 EcoRV/NotI을 사용하여 pcDNA5/FRT/TO (Invitrogen)으로 클론되었다.
세포배양 및 형질감염
HeLa 세포를 5% CO2, 37℃에서 4.5 g/l 글루토오스, 10% 열-비화성된 ㅅ소소태아혈청, 2 mM L-글루타민(glutamine), 및 1 mM 소듐 피루베이트(sodium pyruvate) (Sigma-Aldrich로부터 모두)로 보충된 Dulbecco’s Modified Eagle’s 배지(DMEM)에서 자라게 하였다. 루시퍼리제 리포터 분석을 위하여, 7000 HeLa 세포/웰을 96 웰 플레이트로 분주하였다. 다음날, 공동-형질감염은 제작사의 지침에 따라 에펙탄 형질감염 시약(Effectene Transfection Reagent)(Qiagen)을 사용하여 수행되었다. 인공전사인자 및 루시퍼라제를 위한 코딩하는 플라스미드 미디 제조(midi preparations)는 3:1 비율로 사용되었다. 배지는 형질감염 후 6 시간 및 24 시간에 신선한 DMEM의 웰 당 100 ㎕로 교체되었다.
Flp - InTm T- RexTM 293 발현 세포주의 발생 및 유지
안정적, 테트라사이클린 유도 Flp-InTm T-RexTM293 발현 세포주는 Flp 제조합효소(Recombinase)-매개된 통합에 의해 생성되었다. Flp-InTm T-RexTM 코어 키트를 사용하여, Flp-InTm T-RexTM 숙주세포주는 pFRT/lacZeo 표적부위 벡터 및 pcDNA6/TR 벡터를 형질감염하여 생성되었다. 유도 293 발현 세포주의 생성을 위하여, 관심있는 유전자를 포함하는 pcDNA5/FRT/TO 발현 벡터는 Flp-InTm T-RexTM 숙주세포주에서 FRT 부위의 Flp 제조합효소(Recombinase)-매개된 DNA 재조합을 경유하여 통합된다. 안정적 Flp-InTm T-RexTM발현 세포주는 (DMEM; 10% Tet-FBS; 2mM glut아민(amine); 15㎍/ml blasticidine 및 100㎍/ml hygromycin)을 포함하는 선별 배지에서 유지되었다. 유전자 발현 유도를 위하여 테트라사이클린이 1 ㎍/ml의 최종 농도에 첨가되었다.
혼합된 루시퍼라제 / SEAP 프로모터 활성 분석
HeLa 세포는 인공전사인자발현 구조 및 OPA1 프로모터의 대조군 하에서 배출된 가우시아(Gaussia)루시퍼라제 및 필수 구성요소 CMV 프로모터(Gaussia luciferase Glow Assay Kit, Pierce; SEAP 리포터 유전자분석 화학 발광(chemiluminescent), Roche)의 대조군 하에서 배출된 알칼린 포스파타제를 운반하는 플라스미드로 공동-형질감염된다. 형질감염 이후 2일에, 세포배양 상등액을 수득하고 루시퍼라제활성 및 SEAP 활성을 각각 가우시아 루시퍼라제 글로우 분석키트(Thermo Scientific)및 SEAP 리포터 유전자 분석(Roche)을 사용하여 측정하였다. 세린(serine) 잔기로 교환된 징크핑거 도메인에 있는 모든 시스테인 잔기로 비활성 인공전사인자를 위하여 발현 플라스미드의 공동-형질감염을 대조군으로서 수행하였다. 루시퍼라제 활성을 SEAP 활성에 정상화(notmalized)시켰고 대조군의 퍼센트로서 발현시켰다.
정량적 상온- PCR의 유전자 발현수준의 결정
총 RNA를 제작사의 지침에 따라 RNeasy 플러스 미니키트(Qiagen, Hilden, Germany)를 사용하여 세포로부터 분리하였다. 냉동된 세포 펠렛을 10 ㎕/ml ß-머캅토에탄올을 포함하는 RLT 플러스 용해 버퍼로 재현탁하였다. QIAshredder 스핀 컬럼을 사용하여 균질화한 후에, 총 용해물을 gDNA 제거 스핀 컬럼으로 이동시키고 게놈의 DNA를 제거하였다. 70% 에탄올의 하나의 부피를 첨가하고 총 용해물을 RNeasy 스핀 컬럼으로 이동시켰다. 몇가지 세척 단계 후에, RNA를 30 ㎕ RNase 없는 물의 최종 부피로 용출하였다. RNA를 추가적으로 사용할 때까지 -80℃에서 저장하였다. cDNA의 합성을 제작사의 지침에 따라 높은 용량 cDNA 역전사 키트(High Capacity cDNA Reverse Transcription Kit; Applied Biosystems, Branchburg, New Jersey, USA)를 사용하여 수행하였다. cDNA 합성은 2 ㎕ 10x 버퍼, 0.8 ㎕ 25x dNTP 혼합물, 2 ㎕ 10x 상온 무작위 프라이머(Random Primers), 1 ㎕ 멀티스크라이브 역전사효소(Multiscribe Reverse Transcriptase) 및 4.2 ㎕ H2O를 포함하는 20 ㎕의 총 반응 부피에서 수행되었다. 10 ㎕ RNA의 최종 부피를 첨가하고 반응을 하기의 조건하에서 수행하였다: 25℃에서 10 분, 이어서 37℃에서 2시간 및 85℃에서 5 분의 최종 단계. 정량적 PCR을 1 ㎕ 20x TaqMan Gter Mix, 10.0 ㎕ TaqMan® Universal PCR Master Mix (both Applied Biosystems, Branchburg, New Jersey, USA) 및 8 ㎕ H2O를 포함하는 총 반응 부피의 20 ㎕에서 수행하였다. 각각 반응을 위하여, 1 ㎕의 cDNA를 첨가하였다. qPCR을 하기의 조건하에서 ABI PRISM 7000 서열 검출 시스템(Applied Biosystems, Branchburg, New Jersey, USA)을 사용하여 수행하였다: 50℃에서 2 분동안 시작 단계에 이어서 95℃에서 10 분동안 첫번째 변성(denaturation) 및 95℃에서 15 초 및 60℃에서 1 분의 40 사이클로 구성되는 추가 단계로 있다.
박테리아 발현을 위한 인공전사인자의 클로닝
인공전사인자를 암호화하는 DNA 단편을 인공전사인자와 TAT 단백질 형질도입(transduction)도메인 사이의 His6-태그된 융합 단백질로서 E. coli에서 발현을 위하여 pET41a+ (Novagen)에 기반한 박테리아 발현벡터 pAN983(서열번호:82)로 EcoRV/NotI을 가지는 표준 절차를 사용하여 클론하였다.
OPA1을 표적하는 BL21(DE3)과 같은 적절한 E. coli 숙주세포에서 변환가능한 인공전사인자의 박테리아 생성을 위한 발현 구조체는 pAN1964 (서열번호:83), pAN2053 (서열번호:84), pAN2055 (서열번호:85), pAN2057 (서열번호:86), pAN2059 (서열번호:87), pAN2061 (서열번호:88), 및 pAN2063 (서열번호:89)이다.
인공전사인자 단백질의 생성
주어진 인공전사인자를 위한 발현플라스미드를 가지는 형질전화된 E. coli BL21(DE3)를 OD600이 0.8과 1사이가 도달될 때까지 100 μM ZnCl2로 보충된 1 l LB 배지에서 자라게 하였고, 2 시간 동안 1 mM IPTG로 유도(induction)하였다. 박테리아를 원심분리로 수득하였고, 박테리아 용해물을 초음파 분해(sonication)로 준비하였고, 인클루젼 바디(inclusion bodies)를 정제하였다. 이를 위하여, 인클루젼 바디를 원심분리(5000g, 4℃, 15 분)로 수득하였고 결합버퍼 (50 mM 헤페스, 500 mM NaCl, 10 mM 이미다졸; pH 7.5)의 20 ml로 3회 세척하였다. 정제된 인클루젼 바디를 결합버퍼 A (50 mM 헤페스, 500 mM NaCl, 10 mM 이미다졸, 6M GuHCl; pH 7.5)의 30 ml에서 1시간 동안 얼음에서 용해시켰다. 용해시 인클루젼 바디를 4℃ 및 13'000 g에서 40 분 동안 원심분리하였도 0.45 μm PVDF 필터를 통하여 여과하였다. His-태그된 인공전사인자를 결합버퍼 A 및 용리 버퍼 B (50 mM 헤페스, 500 mM NaCl, 500 mM 이미다졸, 6M GuHCl; pH 7.5)를 사용하여 Aktaprime FPLC (GEHealthcare) 상에서 His-Trap 컬럼을 사용하여 정제하였다. 정제된 인공전사인자를 포함하는 분획은 풀(pool)되었고 SID 도메인을 포함하는 인공전사인자 경우에 버퍼 S (50 mM Tris-HCl, 500 mM NaCl, 200 mM 아르기닌(arginine), 100 μM ZnCl2, 5 mM GSH, 0.5 mM GSSG, 50% 글리세롤; pH 7.5)에 대하여, 또는 인공전사인자를 포함하는 KRAB 도메인을 위하여 버퍼 K (50 mM Tris-HCl, 300 mM NaCl, 500 mM 아르기닌(arginine), 100 μM ZnCl2, 5 mM GSH, 0.5 mM GSSG, 50% 글리세롤; pH 8.5)에 대하여 4℃ 밤사이에서 투석되었다. 투석에 이어서, 단백질 시료를 4℃에서 30 분 동안 14'000 rpm에서 원심분리하였고 0.22 μm Millex-GV filter tips (Millipore)를 사용하여 멸균 여과하였다. VP64 활성도메인을 포함하는 인공전사인자에 대하여, 단백질을 제작사의 추천에 따른 His-Bond Ni-NTA 레진(Novagen)을 사용하여 가용성 분획(결합버퍼: 50 mM NaPO4 pH 7.5, 500 mM NaCl, 10 mM 이미다졸; 용리 버퍼 50 mM 헤페스 pH 7.5, 500 mM NaCl, 500 mM 이미다졸)으로부터 생성되었다. 단백질을 VP64-버퍼 (550 mM NaCl pH 7.4, 400 mM 아르기닌(arginine), 100 μM ZnCl2)에 대하여 투석하였다.
ELDIA (효소-링크된 DNA 상호작용 분석)을 사용하여 인공전사인자의 DNA 결합활성의 결정
BSA 미리-차단된 니켈 코팅된(pre-blocked nickel coated) 플레이트 (Pierce)를 세척 버퍼 (25 mM Tris/HCl pH 7.5, 150 mM NaCl, 0.1% BSA, 0.05% 트윈-20)로 3 회 세척하였다. 플레이트를 저장(storage) 버퍼에서 포화(saturating) 조건(50 pmol/웰) 하에서 정제된 인공전사인자로 코팅하였고 약한 쉐이크(slight shake)로 상온에서 1 시간 배양하였다. 3번의 세척 단계 후에, 60 bp프로모터 서열을 포함하는 애닐된, 바이오티닐된 올리고의 1x 10-12 내지 5x 10-7 M를 상온에서 1시간 동안 인공전사인자와 결합된 비특정한 경쟁체(competitor)(salmon sperm, Sigma로부터 0.1 mg/ml ssDNA)의 존재에서 결합버퍼 (10 mM Tris/HCl pH 7.5, 60 mM KCl, 1 mM DTT, 2% 글리세롤, 5 mM MgCl2 및 100 μM ZnCl2)에서 배양하였다. 세척(5 회)후에, 웰은 상온에서 30 분 동안 3% BSA로 차단되었다. 항-스트렙타비티딘-HRP(Anti-streptavidin-HRP)을 상온에서 1 시간 동안 결합버퍼에서 첨가하였다. 5 세척 단계 후에, TMB 기질(Sigma)을 첨가하였고 상온에서 2 내지 30 분동안 배양하였다. 반응을 TMB 스탑(stop) 용액 (Sigma)의 첨가로 멈추었고 시료 소멸을 450 nm에서 읽었다. 리간드 결합 키네틱스의 데이타 분석을 Hill에 따른 Sigma Plot V8.1을 사용하여 수행하였다.
단백질 형질도입(transduction)
약 80 % 확산(confluency)로 자란 세포를 37℃에서 OptiMEM 또는 성장 배지로 매 24 시간마다 인공전사인자의 선택적 첨가로 2 시간 내지 120 시간동안 처리된 0.01 내지 1 μM 인공전사인자 또는 목(mock)으로 처리되었다. 선택적으로, 10-500 μM ZnCl2을 성장 배지에 첨가하였다. 면역형광을 위하여, 세포를 PBS로 1회 세척하였고, 트립신하였고 추가 시험을 위하여 유리 커버 슬립위에 분주하였다.
면역형광
세포를 4% 파라포름알데하이드로 고정하였고, 15 분동안 0.15 % Triton X-100로 처리하였고, 10% BSAPBS로 차단하였고 마우스 항-HA항체 (1:500, H9658, Sigma) 또는 마우스 항-myc (1:500, M5546, Sigma)로 밤사이 배양하였다. 시료를 PBS/1% BSA로 3회 세척하였고, Alexa Fluor 546 (1:1000, Invitrogen)에 결합된 고우트 항-마우스 항체로 배양하였고 DAPI(3 분동안 1 mg/ml의 1:1000, Sigma)를 사용하여 대비사용하였다. 시료를 형광 현미경을 사용하여 분석되었다.
웨스턴 블랏팅
단백질 수준을 측정하는 동안, 세포를 RIPA 버퍼 (Pierce)를 사용하여 용해하였고 단백질 용해물을 래밀리 시료(Laemmli sample) 버퍼로 혼합하였다. 단백질을 그들의 크기에 따라 SDS-PAGE에 의해 분리되었고 전기 블랏팅(electroblotting)을 사용하여 니트로셀룰로오스(nitrocellulose) 막으로 이동시켰다. 단백질 검출을 마우스 또는 래빗에서 일으킨 특정한 1차 항체를 사용하여 수행하였다. 1차 항체의 검출을 호스래디쉬 퍼옥시다제(horseradish peroxidase) 및 루미네센스(luminescence)-기반 검출 (ECL 플러스, Pierce)에 결합된 2차 항체 또는 적외선 레이저 스캐너(infrared laser scanner)를 사용하여 검출되고 정량되는 DyLight700 또는 DyLight800 형광에 결합된 2차 항체에 의해 수행하였다.
미토콘드리아 기능 측정
유동 세포 측정분석을 위하여, 처리된 세포를 10 mM EDTA/PBS로 수득하였다. Mock 처리된 세포를 대조군으로 사용하였다. 미토콘드리아 막 전위를 측정하기 위하여, 세포를 FACS 버퍼 P (PBS, 5 mM EDTA, 0.5% (w/v) BSA, 1 ㎍/ml 4′,6-디아미디노-2-페닐인돌 디하이드로클로라이드(4′,6-diamidino-2-phenylindole dihydrochloride) (DAPI, Sigma), 10 nM 테트라메틸로다민 에틸에스테르(tetramethylrhodamine ethylester) (TMRE, Sigma))에서 재현탁하였고 분석하기 전에 37℃에서 30 분 동안 배양하였다. 미토콘드리아 막 전위를 소멸하기 위하여 50 μM 카보닐 3-클로로페닐하이드라존(carbonyl cyanide 3-chlorophenylhydrazone)(CCCP, Sigma)으로 처리는 대조군으로서 수행하였다. 미토콘드리아 질량의 측정을 위하여, 세포를 FACS 버퍼 M (PBS, 5 mM EDTA, 0.5% (w/v) BSA, 1 ㎍/ml DAPI 및 100 nM MitoTracker green FM (Invitrogen))에서 재현탁하고 분석 전에 37 ℃에서 30 분 동안 배양하였다. 미토콘드리아 ROS 측정을 위하여, 세포를 FACS 버퍼 R (PBS, 5 mM EDTA, 0.5% BSA, 1 ㎍/ml DAPI 및 5 μM MitoSOX (Invitrogen)에서 재현탁하였고, 37℃에서 10 분 동안 배양하였고, PBS로 세척하였고, FACS 버퍼 R2 (PBS, 5 mM EDTA, 0.5% (w/v) BSA)에서 재현탁하였다. 유동 세포 측정분석을 lowJo 소프트웨어(Tree Star Inc.)을 사용하여 CyAnADP (Dako) 상에서 수행하였다.
세포사멸( apoptotic ) 유도 측정
세포를 포스페이트-버퍼된 살린(phospate-buffered saline)(PBS)에서 4% EM-등급 파라포름알데하이드(Pierce, 28908)으로 상온에서 30 분 동안 고정하였다. 이후, 세포를 상온에서 15 분 동아 PBS에서 0.15% (v/v) 트리톤 X-100으로 투과시켰고, 이어서 상온에서 1 시간 동안 PBS에서 10% (w/v) BSA으로 차단시켰다. 시료를 차단 버퍼(blocking buffer)에서 희석된 마우스 항-사이토크롬 c(cytochrome c)항체 (BD Biosciences, 556432, 1:1000)으로 4℃에서 밤사이 배양하였다. 세포를 차단 버퍼로 15 분 동안 3회 세척하였고 이후 Alexa Fluor 546-결합된 고우트 항-마우스 IgG 항체 (Invitrogen)로 상온에서 1시간 동안 배양하였다. 세포사멸의 측정으로서 사이토크롬 c(cytochrome c)배출을 맹인 관찰자(blinded observer)에 의해 형광 현미경으로 분석하였다. Mock 처리된 세포를 대조군으로서 수행하였다.
<110> Aliophtha AG <120> Artificial transcription factors for the treatment of diseases caused by OPA1 haploinsufficiency <130> P3032PC00 <150> EP13162189.8 <151> 2013-04-03 <160> 89 <170> PatentIn version 3.5 <210> 1 <211> 13 <212> PRT <213> herpes simplex virus 7 <400> 1 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser 1 5 10 <210> 2 <211> 55 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 2 Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser 1 5 10 15 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 20 25 30 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 35 40 45 Asp Leu Asp Met Leu Ile Asn 50 55 <210> 3 <211> 102 <212> PRT <213> Homo sapiens <400> 3 Lys Gly Phe Gly Ala Phe Glu Arg Ser Ile Leu Thr Gln Ile Asp His 1 5 10 15 Ile Leu Met Asp Lys Glu Arg Leu Leu Arg Arg Thr Gln Thr Lys Arg 20 25 30 Ser Val Tyr Arg Val Leu Gly Lys Pro Glu Pro Ala Ala Gln Pro Val 35 40 45 Pro Glu Ser Leu Pro Gly Glu Pro Glu Ile Leu Pro Gln Ala Pro Ala 50 55 60 Asn Ala His Leu Lys Asp Leu Asp Glu Glu Ile Phe Asp Asp Asp Asp 65 70 75 80 Phe Tyr His Gln Leu Leu Arg Glu Leu Ile Glu Arg Lys Thr Ser Ser 85 90 95 Leu Asp Pro Asn Asp Gln 100 <210> 4 <211> 31 <212> PRT <213> Homo sapiens <400> 4 Pro Gly Leu Pro Asn Gly Leu Leu Ser Gly Asp Glu Asp Phe Ser Ser 1 5 10 15 Ile Ala Asp Met Asp Phe Ser Ala Leu Leu Ser Gln Ile Ser Ser 20 25 30 <210> 5 <211> 48 <212> PRT <213> Homo sapiens <400> 5 Pro Tyr Thr Pro Asn Leu Pro His His Gln Asn Gly His Leu Gln His 1 5 10 15 His Pro Pro Met Pro Pro His Pro Gly His Tyr Trp Pro Val His Asn 20 25 30 Glu Leu Ala Phe Gln Pro Pro Ile Ser Asn His Pro Ala Pro Glu Tyr 35 40 45 <210> 6 <211> 100 <212> PRT <213> Homo sapiens <400> 6 Pro Pro His Leu Asn Pro Gln Asp Pro Leu Lys Asp Leu Val Ser Leu 1 5 10 15 Ala Cys Asp Pro Ala Ser Gln Gln Pro Gly Pro Leu Asn Gly Ser Gly 20 25 30 Gln Leu Lys Met Pro Ser His Cys Leu Ser Ala Gln Met Leu Ala Pro 35 40 45 Pro Pro Pro Gly Leu Pro Arg Leu Ala Leu Pro Pro Ala Thr Lys Pro 50 55 60 Ala Thr Thr Ser Glu Gly Gly Ala Thr Ser Pro Thr Ser Pro Ser Tyr 65 70 75 80 Ser Pro Pro Asp Thr Ser Pro Ala Asn Arg Ser Phe Val Gly Leu Gly 85 90 95 Pro Arg Asp Pro 100 <210> 7 <211> 68 <212> PRT <213> Homo sapiens <400> 7 Ala Asp Phe Gln Pro Pro Tyr Phe Pro Pro Pro Tyr Gln Pro Ile Tyr 1 5 10 15 Pro Gln Ser Gln Asp Pro Tyr Ser His Val Asn Asp Pro Tyr Ser Leu 20 25 30 Asn Pro Leu His Ala Gln Pro Gln Pro Gln His Pro Gly Trp Pro Gly 35 40 45 Gln Arg Gln Ser Gln Glu Ser Gly Leu Leu His Thr His Arg Gly Leu 50 55 60 Pro His Gln Leu 65 <210> 8 <211> 112 <212> PRT <213> Homo sapiens <400> 8 Asn Arg Thr Val Ser Gly Gly Gln Tyr Val Val Ala Ala Ala Pro Asn 1 5 10 15 Leu Gln Asn Gln Gln Val Leu Thr Gly Leu Pro Gly Val Met Pro Asn 20 25 30 Ile Gln Tyr Gln Val Ile Pro Gln Phe Gln Thr Val Asp Gly Gln Gln 35 40 45 Leu Gln Phe Ala Ala Thr Gly Ala Gln Val Gln Gln Asp Gly Ser Gly 50 55 60 Gln Ile Gln Ile Ile Pro Gly Ala Asn Gln Gln Ile Ile Thr Asn Arg 65 70 75 80 Gly Ser Gly Gly Asn Ile Ile Ala Ala Met Pro Asn Leu Leu Gln Gln 85 90 95 Ala Val Pro Leu Gln Gly Leu Ala Asn Asn Val Leu Ser Gly Gln Thr 100 105 110 <210> 9 <211> 143 <212> PRT <213> Homo sapiens <400> 9 Gln Gly Gln Thr Pro Gln Arg Val Ser Gly Leu Gln Gly Ser Asp Ala 1 5 10 15 Leu Asn Ile Gln Gln Asn Gln Thr Ser Gly Gly Ser Leu Gln Ala Gly 20 25 30 Gln Gln Lys Glu Gly Glu Gln Asn Gln Gln Thr Gln Gln Gln Gln Ile 35 40 45 Leu Ile Gln Pro Gln Leu Val Gln Gly Gly Gln Ala Leu Gln Ala Leu 50 55 60 Gln Ala Ala Pro Leu Ser Gly Gln Thr Phe Thr Thr Gln Ala Ile Ser 65 70 75 80 Gln Glu Thr Leu Gln Asn Leu Gln Leu Gln Ala Val Pro Asn Ser Gly 85 90 95 Pro Ile Ile Ile Arg Thr Pro Thr Val Gly Pro Asn Gly Gln Val Ser 100 105 110 Trp Gln Thr Leu Gln Leu Gln Asn Leu Gln Val Gln Asn Pro Gln Ala 115 120 125 Gln Thr Ile Thr Leu Ala Pro Met Gln Gly Val Ser Leu Gly Gln 130 135 140 <210> 10 <211> 95 <212> PRT <213> Homo sapiens <400> 10 Asp Leu Gln Gln Leu Gln Gln Leu Gln Gln Gln Asn Leu Asn Leu Gln 1 5 10 15 Gln Phe Val Leu Val His Pro Thr Thr Asn Leu Gln Pro Ala Gln Phe 20 25 30 Ile Ile Ser Gln Thr Pro Gln Gly Gln Gln Gly Leu Leu Gln Ala Gln 35 40 45 Asn Leu Leu Thr Gln Leu Pro Gln Gln Ser Gln Ala Asn Leu Leu Gln 50 55 60 Ser Gln Pro Ser Ile Thr Leu Thr Ser Gln Pro Ala Thr Pro Thr Arg 65 70 75 80 Thr Ile Ala Ala Thr Pro Ile Gln Thr Leu Pro Gln Ser Gln Ser 85 90 95 <210> 11 <211> 63 <212> PRT <213> Homo sapiens <400> 11 Gln Leu Ala Gly Asp Ile Gln Gln Leu Leu Gln Leu Gln Gln Leu Val 1 5 10 15 Leu Val Pro Gly His His Leu Gln Pro Pro Ala Gln Phe Leu Leu Pro 20 25 30 Gln Ala Gln Gln Ser Gln Pro Gly Leu Leu Pro Thr Pro Asn Leu Phe 35 40 45 Gln Leu Pro Gln Gln Thr Gln Gly Ala Leu Leu Thr Ser Gln Pro 50 55 60 <210> 12 <211> 90 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 12 Asn Leu Phe Gln Leu Pro Gln Gln Thr Gln Gly Ala Leu Leu Thr Ser 1 5 10 15 Gln Pro Asn Leu Phe Gln Leu Pro Gln Gln Thr Gln Gly Ala Leu Leu 20 25 30 Thr Ser Gln Pro Asn Leu Phe Gln Leu Pro Gln Gln Thr Gln Gly Ala 35 40 45 Leu Leu Thr Ser Gln Pro Asn Leu Phe Gln Leu Pro Gln Gln Thr Gln 50 55 60 Gly Ala Leu Leu Thr Ser Gln Pro Asn Leu Phe Gln Leu Pro Gln Gln 65 70 75 80 Thr Gln Gly Ala Leu Leu Thr Ser Gln Pro 85 90 <210> 13 <211> 91 <212> PRT <213> Homo sapiens <400> 13 Pro Pro Ser Thr Gly Asn Ser Ala Ser Leu Ser Leu Pro Leu Val Leu 1 5 10 15 Gln Pro Gly Leu Ser Glu Pro Pro Gln Pro Leu Leu Pro Ala Ser Ala 20 25 30 Pro Ser Ala Pro Pro Pro Ala Pro Ser Leu Gly Pro Gly Ser Gln Gln 35 40 45 Ala Ala Phe Gly Asn Pro Pro Ala Leu Leu Gln Pro Pro Glu Val Pro 50 55 60 Val Pro His Ser Thr Gln Phe Ala Ala Asn His Gln Glu Phe Leu Pro 65 70 75 80 His Pro Gln Ala Pro Gln Pro Ile Val Pro Gly 85 90 <210> 14 <211> 111 <212> PRT <213> Homo sapiens <400> 14 Met Ala Thr Arg Val Leu Ser Met Ser Ala Arg Leu Gly Pro Val Pro 1 5 10 15 Gln Pro Pro Ala Pro Gln Asp Glu Pro Val Phe Ala Gln Leu Lys Pro 20 25 30 Val Leu Gly Ala Ala Asn Pro Ala Arg Asp Ala Ala Leu Phe Pro Gly 35 40 45 Glu Glu Leu Lys His Ala His His Arg Pro Gln Ala Gln Pro Ala Pro 50 55 60 Ala Gln Ala Pro Gln Pro Ala Gln Pro Pro Ala Thr Gly Pro Arg Leu 65 70 75 80 Pro Pro Glu Asp Leu Val Gln Thr Arg Cys Glu Met Glu Lys Tyr Leu 85 90 95 Thr Pro Gln Leu Pro Pro Val Pro Ile Ile Pro Glu His Lys Lys 100 105 110 <210> 15 <211> 88 <212> PRT <213> Homo sapiens <400> 15 Met Ala Leu Ser Glu Pro Ile Leu Pro Ser Phe Ser Thr Phe Ala Ser 1 5 10 15 Pro Cys Arg Glu Arg Gly Leu Gln Glu Arg Trp Pro Arg Ala Glu Pro 20 25 30 Glu Ser Gly Gly Thr Asp Asp Asp Leu Asn Ser Val Leu Asp Phe Ile 35 40 45 Leu Ser Met Gly Leu Asp Gly Leu Gly Ala Glu Ala Ala Pro Glu Pro 50 55 60 Pro Pro Pro Pro Pro Pro Pro Ala Phe Tyr Tyr Pro Glu Pro Gly Ala 65 70 75 80 Pro Pro Pro Tyr Ser Ala Pro Ala 85 <210> 16 <211> 11 <212> PRT <213> Human immunodeficiency virus <400> 16 Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg 1 5 10 <210> 17 <211> 1000 <212> DNA <213> Homo sapiens <400> 17 gaaatttggg aggggagcca tcaaagaagc ctgggagcag cagttccagg gaaaaaggag 60 aatgtgatgg ccagagagcc aaaagaaaaa gtagttgaag gagtgctcag cactaggcat 120 ctgaactgaa tgctgtggca ggctcactgg ccacaaacaa tagggagctg gtggaggcct 180 tgacgaggac catttcaaca aactggtggg cttaaaatcc ggaagaaaca gttgaacaaa 240 tcattttgac gccttttata aaccacacaa gcttattcca aacccgttac tggcctaact 300 gatttaagtc cctttcccat ctgatcctca gagattctaa gggacttagc ctatccatga 360 ctcttcgtcc tgcttctcac ctcccatgat tgccctaacg atgtgaaagt gctttcaaac 420 aaagatgccc aagaaagaag gtaggcaaat gtgcaagcat tagtttgtag tacgctatta 480 ctgtatttca ccttgcactc tctagtttcc ttcgtgctcc ctcaatatcc aactcttaat 540 aaattcatgg ctcccggtga gcattcatca attctcattc cacgccttta gcccttcccg 600 ttcccgccca actctcgctc cctcccctgg ccaaatctct aacctgcaag gctaattccg 660 aattccaaat cggaagcaag agggcggggc cccgtgagag gcgatggatt gctccagtcc 720 gttcccgacg cactgtgcgc atgcgctggt cctccgcgga ccgttcgtgc tgcccgccta 780 gaaagggtga agtggttgtt tccgtgacgg actgagtacg ggtgcctgtc aggctcttgc 840 ggaagtccat gcgccattgg gagggcctcg gccgcggctc tgtgcccttg ctgctgaggg 900 ccacttcctg ggtcattcct ggaccgggag ccgggctggg gctcacacgg gggctcccgc 960 gtggccgtct cggcgcctgc gtgacctccc cgccggcggg 1000 <210> 18 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 18 Pro Val Arg Arg Pro Arg Arg Arg Arg Arg Arg Lys 1 5 10 <210> 19 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 19 Thr His Arg Leu Pro Arg Arg Arg Arg Arg Arg Lys 1 5 10 <210> 20 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 20 Arg Arg Arg Arg Arg Arg Arg Arg Arg 1 5 <210> 21 <211> 16 <212> PRT <213> Drosophila melanogaster <400> 21 Arg Gln Ile Leu Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys 1 5 10 15 <210> 22 <211> 18 <212> DNA <213> Homo sapiens <400> 22 gaaaaagtag ttgaagga 18 <210> 23 <211> 18 <212> DNA <213> Homo sapiens <400> 23 gtagttgaag gagtgctc 18 <210> 24 <211> 18 <212> DNA <213> Homo sapiens <400> 24 gacctccccg ccggcggg 18 <210> 25 <211> 20 <212> DNA <213> Homo sapiens <400> 25 ctcttgcgga agtccatgcg 20 <210> 26 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 26 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 1 5 10 15 Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 115 120 125 Arg Ala Asn Leu Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu 145 150 155 160 Val Arg His Gln Arg Thr His Thr 165 <210> 27 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 27 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser 1 5 10 15 Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr 115 120 125 Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu 145 150 155 160 Val Arg His Gln Arg Thr His Thr 165 <210> 28 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 28 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 1 5 10 15 Asn Asp Thr Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr 115 120 125 Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu 145 150 155 160 Val Arg His Gln Arg Thr His Thr 165 <210> 29 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 29 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser 1 5 10 15 Lys Lys Ala Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 115 120 125 Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu 145 150 155 160 Val Arg His Gln Arg Thr His Thr 165 <210> 30 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 30 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser 1 5 10 15 Lys Lys His Leu Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 115 120 125 Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu 145 150 155 160 Val Arg His Gln Arg Thr His Thr 165 <210> 31 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 31 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 1 5 10 15 Pro Gly Ala Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 115 120 125 Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu 145 150 155 160 Ala Glu His Gln Arg Thr His Thr 165 <210> 32 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 32 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 1 5 10 15 Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr 115 120 125 Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu 145 150 155 160 Val Arg His Gln Arg Thr His Thr 165 <210> 33 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 33 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 1 5 10 15 Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr 115 120 125 Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu 145 150 155 160 Val Arg His Gln Arg Thr His Thr 165 <210> 34 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 34 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 1 5 10 15 Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr 115 120 125 Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu 145 150 155 160 Ala Arg His Gln Arg Thr His Thr 165 <210> 35 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 35 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 1 5 10 15 Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu 35 40 45 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Arg Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr 115 120 125 Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu 145 150 155 160 Val Arg His Gln Arg Thr His Thr 165 <210> 36 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 36 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 1 5 10 15 Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys 35 40 45 Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 115 120 125 Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys 145 150 155 160 Arg Ala His Gln Arg Thr His Thr 165 <210> 37 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 37 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 1 5 10 15 Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu 35 40 45 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 115 120 125 Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu 145 150 155 160 Thr Glu His Gln Arg Thr His Thr 165 <210> 38 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 38 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 1 5 10 15 Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu 35 40 45 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 115 120 125 Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu 145 150 155 160 Ala Glu His Gln Arg Thr His Thr 165 <210> 39 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 39 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 1 5 10 15 Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu 35 40 45 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 115 120 125 Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu 145 150 155 160 Ala Glu His Gln Arg Thr His Thr 165 <210> 40 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 40 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 1 5 10 15 Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu 35 40 45 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 115 120 125 Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu 145 150 155 160 Thr Glu His Gln Arg Thr His Thr 165 <210> 41 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 41 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 1 5 10 15 Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys 35 40 45 Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 115 120 125 Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu 145 150 155 160 Thr Glu His Gln Arg Thr His Thr 165 <210> 42 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 42 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 1 5 10 15 Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu 35 40 45 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 115 120 125 Pro Gly Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu 145 150 155 160 Thr Glu His Gln Arg Thr His Thr 165 <210> 43 <211> 168 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 43 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 1 5 10 15 Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 20 25 30 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys 35 40 45 Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 50 55 60 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln 65 70 75 80 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 85 90 95 Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr 100 105 110 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr 115 120 125 Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 130 135 140 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Asn Ser Thr Leu 145 150 155 160 Thr Glu His Gln Arg Thr His Thr 165 <210> 44 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 44 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu 20 25 30 Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Arg Ala Asn Leu 130 135 140 Arg Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu Val Arg His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 45 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 45 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu 20 25 30 Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 46 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 46 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Thr Leu 20 25 30 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 47 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 47 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys Ala Leu 20 25 30 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 100 105 110 Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 48 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 48 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu 20 25 30 Ala Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 49 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 49 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Arg Ser Asp Lys Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 100 105 110 Pro Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu 130 135 140 Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu Ala Glu His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 50 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 50 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 100 105 110 Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu 130 135 140 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu Val Arg His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 51 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 51 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 100 105 110 Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu 130 135 140 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 52 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 52 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 100 105 110 Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu 130 135 140 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 53 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 53 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg 100 105 110 Ser Asp Lys Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu 130 135 140 Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 54 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 54 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 55 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 55 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Thr Glu His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 56 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 56 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu Ala Glu His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 57 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 57 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Ser Ser Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys His Leu Ala Glu His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 58 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 58 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Thr Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Lys Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu Thr Glu His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 59 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 59 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Gln Arg Ala His Leu Glu Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp 100 105 110 Cys Arg Asp Leu Ala Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser Gly Asp Leu 130 135 140 Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu Thr Glu His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 60 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 60 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asn Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Thr Thr Gly Ala Leu Thr Glu His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln 100 105 110 Ser Gly Asp Leu Arg Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly Asn Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu Thr Glu His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 61 <211> 276 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 61 Met Pro Lys Lys Lys Arg Lys Val Gly Leu Glu Pro Gly Glu Lys Pro 1 5 10 15 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Asp Leu 20 25 30 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 35 40 45 Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg Thr Cys Arg Ala His Gln 50 55 60 Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 65 70 75 80 Ser Phe Ser Arg Ser Asp Asp Leu Val Arg His Gln Arg Thr His Thr 85 90 95 Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr 100 105 110 Ser Gly Ser Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro 115 120 125 Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu Leu 130 135 140 Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro 145 150 155 160 Glu Cys Gly Lys Ser Phe Ser Gln Asn Ser Thr Leu Thr Glu His Gln 165 170 175 Arg Thr His Thr Gly Gly Gly Ser Gly Gly Ser Glu Phe Gly Arg Ala 180 185 190 Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu 195 200 205 Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe 210 215 220 Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp 225 230 235 240 Met Leu Ile Asn Gly Ser Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 245 250 255 Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Glu Gln Lys Leu Ile Ser 260 265 270 Glu Glu Asp Leu 275 <210> 62 <211> 7 <212> PRT <213> Simian virus 40 <400> 62 Pro Lys Lys Lys Arg Lys Val 1 5 <210> 63 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 63 Gly Gly Ser Gly Gly Ser 1 5 <210> 64 <211> 4513 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 64 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt accgtatacc 420 tcgagcccgg ggaaaagcca tataaatgcc ccgagtgcgg caaatcattc agccaaagta 480 gcaacttagt aagacaccag cgcacccata ccggtaagaa aactagtctt aagctcgagc 540 ccggggaaaa accctataaa tgccccgagt gtggtaagtc attctctcaa agcggggatt 600 taagaagaca ccagagaacc cacaccggta agaaaactag tggcgcgccc tcgagcccgg 660 ggagaaacct tataaatgcc cagaatgcgg gaaatcgttc agtcaaagag cacatttaga 720 aagacatcaa cggacccaca ccggtaagaa aactagtcct aggctcgagc ccggggaaaa 780 accttacaag tgccctgagt gcggcaagag cttctctcaa tcaagttcat tagtaagaca 840 ccagaggact cataccggta agaaaactag tcctcagcct cgagcccggg gagaagcctt 900 ataagtgccc tgagtgtggc aaaagcttca gcgatcctgg aaatttagta agacaccaac 960 gcacccacac cggtaagaaa actagtatgc atctcgagcc cggggaaaaa ccgtataaat 1020 gtcctgagtg cggtaagtct ttttccgact gtagagactt agcgagacac caacgtactc 1080 ataccggtaa aaagactagt tgtacactcg agcccgggga aaaaccgtac aagtgtcctg 1140 agtgcgggaa gagtttctcc gatccgggcc acttagtaag acatcagagg acacataccg 1200 gtaaaaagac tagtttcgaa ctcgagcccg gggagaaacc atacaaatgc cccgagtgtg 1260 gaaagtcatt tagtgatcca ggcgcattag taagacatca gcggacacat accggtaaga 1320 aaactagtga attcctcgag cccggggaga agccatataa atgtcccgag tgtggcaagt 1380 ccttttctag atcagataat ttagtaagac atcagagaac gcacaccggt aaaaagacta 1440 gtcaattgct cgagcccggg gagaagccat acaagtgtcc cgaatgcggg aagtcattct 1500 ccagaagtga cgatttagta agacatcagc gcacgcacac cggtaagaaa actagtccat 1560 ggctcgagcc cggggagaag ccctacaagt gtccagaatg cggaaagagt ttctccagaa 1620 gtgacaaatt agtaagacac cagagaaccc ataccggtaa gaaaactagt catatgctcg 1680 agcccgggga gaagccgtac aagtgccctg aatgtggtaa gtcattttcg agaagtgatg 1740 aattagtaag acaccagcgg actcataccg gtaaaaagac tagtgctagc ctcgagcccg 1800 gggagaagcc ctataaatgt ccagaatgtg gaaagtcctt tagcacgtca gggaacttag 1860 taagacacca gcgaactcat accggtaaga aaactagttt aattaactcg agcccgggga 1920 gaaaccatac aagtgtccag agtgcgggaa aagctttagt acaagcggtg agttagtaag 1980 acaccaacga acacacaccg gtaaaaagac tagtgtttaa acctcgagcc cggggaaaag 2040 ccctacaagt gcccggaatg cggcaagtct tttagcacca gcggacattt agtaagacac 2100 cagagaaccc acaccggtaa aaagactagt ccgcggctcg agcccgggga aaagccctac 2160 aagtgtcctg agtgcggaaa gtctttctcc actagcggtt cattagtaag acaccagagg 2220 acacacaccg gtaaaaagac tagtgcatgc gtcgactgca gaggcctgca tgcaagcttg 2280 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 2340 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 2400 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 2460 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 2520 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 2580 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 2640 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 2700 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 2760 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 2820 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 2880 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 2940 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 3000 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 3060 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 3120 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 3180 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 3240 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 3300 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 3360 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 3420 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 3480 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 3540 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 3600 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 3660 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 3720 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 3780 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 3840 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 3900 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 3960 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 4020 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 4080 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 4140 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 4200 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 4260 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 4320 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 4380 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 4440 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg 4500 aggccctttc gtc 4513 <210> 65 <211> 4442 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 65 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa 420 tgcatctaga tgtatacctc gagcccgggg agaagcccta taaatgccct gaatgcggga 480 aatctttctc ttctaagaag gcactcacag aacaccagcg gacacacacc ggtaaaaaaa 540 ctagtcttaa gctcgagccc ggggaaaagc cctacaagtg ccccgaatgc gggaagtctt 600 ttagtcagag tggaaatctt accgagcacc agagaacaca caccggtaag aagactagtg 660 gcgcgccctc gagcccgggg agaagccata caagtgccct gaatgtggca agtccttttc 720 aagagccgat aacctgacag aacaccaaag gacgcatacc ggtaagaaaa ctagtcctag 780 gctcgagccc ggggagaagc cctataaatg ccctgaatgt ggcaagagct tcagtactag 840 cgggaatctc actgaacatc agcgaactca taccggtaaa aaaactagtc ctcagcctcg 900 agcccgggga aaaaccatac aagtgccctg agtgcggcaa gagttttagt acctcacact 960 ctcttacaga acatcagcga acccacaccg gtaaaaaaac tagtatgcat ctcgagcccg 1020 gggagaaacc atacaaatgt cccgaatgtg gcaagagttt cagcagtaaa aagcatctcg 1080 ctgagcatca gagaactcac accggtaaaa agactagttg tacactcgag cccggggaaa 1140 agccctacaa atgccccgaa tgtggtaagt ctttttctag gaacgacacc ttgacagaac 1200 accagcggac ccacaccggt aagaagacta gtgaattcct cgagcccggg gagaagcctt 1260 ataagtgccc cgaatgtgga aagagtttct ctactaagaa tagcctgacc gagcaccagc 1320 gcactcacac cggtaagaaa actagtcaat tgctcgagcc cggggagaag ccctataaat 1380 gccctgaatg cgggaaatct ttctctcaat caggccacct cacagaacac cagcggacac 1440 acaccggtaa aaaaactagt ccatggctcg agcccgggga gaaaccctat aagtgtcccg 1500 aatgcgggaa atcattctct catacagggc atctgctcga acatcaaagg acgcacaccg 1560 gtaaaaagac tagtcatatg ctcgagcccg gggaaaagcc ttacaaatgc cccgaatgtg 1620 ggaagagttt cagccggtct gataagctga ccgaacacca gagaactcat accggtaaaa 1680 aaactagtgc tagcctcgag cccggggaaa agccctacaa gtgccctgag tgtgggaagt 1740 ccttttcttc aagacgcacg tgccgcgctc accagcggac acataccggt aagaaaacta 1800 gtttaattaa ctcgagcccg gggagaaacc atacaaatgt cccgaatgtg gcaagtcctt 1860 ctcacagaac tctactttga ccgagcatca gagaactcac accggtaaga agactagtcc 1920 gcggctcgag cccggggaaa agccttataa gtgccccgaa tgcggaaaga gcttctcaag 1980 gaatgatgca cttaccgagc atcaaaggac tcataccggt aaaaaaacta gtgcatgctt 2040 cgaactcgag cccggggaaa agccctataa gtgtcccgaa tgcggcaaga gttttagtac 2100 tactggcgca ctcacagaac accagcgcac tcacaccggt aagaaaacta gtgaaagtcc 2160 tctccactga ctgtagcctc caattcactg gagatctgac acaagcttgg cgtaatcatg 2220 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 2280 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 2340 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 2400 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 2460 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 2520 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 2580 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 2640 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 2700 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 2760 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 2820 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 2880 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 2940 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 3000 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 3060 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 3120 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 3180 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 3240 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 3300 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 3360 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 3420 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 3480 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 3540 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 3600 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 3660 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 3720 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 3780 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 3840 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 3900 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 3960 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 4020 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 4080 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 4140 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 4200 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 4260 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 4320 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 4380 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 4440 tc 4442 <210> 66 <211> 4376 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <400> 66 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt accgtatacc 420 tcgagcccgg ggagaagcca tacaaatgcc ctgagtgtgg aaagtcattt agccagcgag 480 ctaatctgcg ggcccaccag cggacccaca ccggtaagaa gactagtctt aagctcgagc 540 ccggggagaa gccatacaaa tgtccagaat gtggaaagtc cttctctgat agtggcaacc 600 tcagagtgca tcagcgaaca cataccggta agaagactag tggcgcgccc tcgagcccgg 660 ggaaaagcca tataagtgcc ctgagtgtgg aaagagcttc agtaggaagg ataaccttaa 720 aaaccaccaa agaacccaca ccggtaagaa gactagtcct aggctcgagc ccggggaaaa 780 gccatataaa tgtcccgagt gcggcaaatc cttctctacc actggcaacc tcacagtgca 840 tcaacggact cacaccggta aaaagactag tcctcagcct cgagcccggg gaaaagccct 900 ataaatgtcc cgagtgcgga aagtcttttt ccagccctgc cgacctgaca cgccaccaac 960 gaacgcacac cggtaagaag actagtatgc atctcgagcc cggggaaaag ccgtacaaat 1020 gtccagagtg tggaaaatcc ttttctgata aaaaggacct gacacggcat cagcgaaccc 1080 acaccggtaa aaagactagt tgtacactcg agcccgggga gaaaccttat aaatgcccag 1140 aatgcggtaa aagtttcagc aggacggata ccttgcggga tcatcagaga acccacaccg 1200 gtaaaaaaac tagtgaattc ctcgagcccg gggaaaaacc atacaagtgc cccgagtgtg 1260 gcaagagctt tagtacccac ctcgacctga ttagacacca gcgcacccac accggtaaga 1320 aaactagtca attgctcgag cccggggaaa agccctataa gtgcccagag tgcgggaaat 1380 cattctcaca gctggcacat cttagagccc accagcggac ccacaccggt aagaagacta 1440 gtccatggct cgagcccggg gagaaaccct ataagtgccc tgaatgcggc aagtctttca 1500 gtgagcggtc acatctccga gagcaccagc gaacgcacac cggtaaaaag actagtcata 1560 tgctcgagcc cggggaaaaa ccctacaagt gccctgagtg tggaaagtca tttagtcgct 1620 ccgaccacct gaccaaccat cagcggactc acaccggtaa gaaaactagt gctagcctcg 1680 agcccgggga gaaaccttac aagtgccccg agtgcggcaa gagtttcagc cacaggacca 1740 ccctgacaaa ccaccagagg acccacaccg gtaaaaagac tagtttaatt aactcgagcc 1800 cggggagaaa ccttataagt gtcctgagtg cggcaaaagt ttctctcaaa agtcctccct 1860 tattgcccat caaaggaccc ataccggtaa gaagactagt gtttaaacct cgagcccggg 1920 gagaagccct ataaatgtcc cgagtgcgga aagtccttct cacggcgcga tgaattgaac 1980 gtccatcaga gaacacacac cggtaaaaaa actagtccgc ggctcgagcc cggggaaaaa 2040 ccttataagt gtcccgagtg cggcaagagt ttcagtcaca aaaacgcact tcagaatcat 2100 cagaggacac ataccggtaa gaaaactagt gcatgcaagc ttggcgtaat catggtcata 2160 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 2220 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 2280 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 2340 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 2400 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 2460 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 2520 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 2580 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 2640 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 2700 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 2760 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 2820 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 2880 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 2940 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 3000 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 3060 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 3120 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 3180 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 3240 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 3300 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 3360 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 3420 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 3480 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 3540 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 3600 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 3660 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 3720 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 3780 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 3840 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 3900 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 3960 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 4020 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 4080 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 4140 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 4200 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 4260 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 4320 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc 4376 <210> 67 <211> 57 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 67 tcgacaggcc caggcggccc tcgaggatat catgatgact agtggccagg ccggccc 57 <210> 68 <211> 57 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 68 aattgggccg gcctggccac tagtcatcat gatatcctcg agggccgcct gggcctg 57 <210> 69 <211> 6699 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 69 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc 60 atatccgcaa tgacaaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga 120 cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatcttcgaa cacacgaaac 180 tttttccttc cttcattcac gcacactact ctctaatgag caacggtata cggccttcct 240 tccagttact tgaatttgaa ataaaaaaag tttgccgctt tgctatcaag tataaataga 300 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt 360 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gctttgcaaa 420 gatggataaa gcggaattaa ttcccgagcc tccaaaaaag aagagaaagg tcgaattggg 480 taccgccgcc aattttaatc aaagtgggaa tattgctgat agctcattgt ccttcacttt 540 cactaacagt agcaacggtc cgaacctcat aacaactcaa acaaattctc aagcgctttc 600 acaaccaatt gcctcctcta acgttcatga taacttcatg aataatgaaa tcacggctag 660 taaaattgat gatggtaata attcaaaacc actgtcacct ggttggacgg accaaactgc 720 gtataacgcg tttggaatca ctacagggat gtttaatacc actacaatgg atgatgtata 780 taactatcta ttcgatgatg aagatacccc accaaaccca aaaaaagaga tctctcgaca 840 ggcccaggcg gccctcgagg atatcatgat gactagtggc caggccggcc caattccaga 900 tctatgaatc gtagatactg aaaaaccccg caagttcact tcaactgtgc atcgtgcacc 960 atctcaattt ctttcattta tacatcgttt tgccttcttt tatgtaacta tactcctcta 1020 agtttcaatc ttggccatgt aacctctgat ctatagaatt ttttaaatga ctagaattaa 1080 tgcccatctt ttttttggac ctaaattctt catgaaaata tattacgagg gcttattcag 1140 aagctttgga cttcttcgcc agaggtttgg tcaagtctcc aatcaaggtt gtcggcttgt 1200 ctaccttgcc agaaatttac gaaaagatgg aaaagggtca aatcgttggt agatacgttg 1260 ttgacacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta 1320 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct 1380 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg 1440 ctcttattga ccacacctct accggcatgc cggtcgaaat tcccctaccc tatgaacata 1500 ttccattttg taatttcgtg tcgtttctat tatgaatttc atttataaag tttatgtaca 1560 aatatcataa aaaaagagaa tctttttaag caaggatttt cttaacttct tcggcgacag 1620 catcaccgac ttcggtggta ctgttggaac cacctaaatc accagttctg atacctgcat 1680 ccaaaacctt tttaactgca tcttcaatgg ccttaccttc ttcaggcaag ttcaatgaca 1740 atttcaacat cattgcagca gacaagatag tggcgatagg gtcaacctta ttctttggca 1800 aatctggagc agaaccgtgg catggttcgt acaaaccaaa tgcggtgttc ttgtctggca 1860 aagaggccaa ggacgcagat ggcaacaaac ccaaggaacc tgggataacg gaggcttcat 1920 cggagatgat atcaccaaac atgttgctgg tgattataat accatttagg tgggttgggt 1980 tcttaactag gatcatggcg gcagaatcaa tcaattgatg ttgaaccttc aatgtaggaa 2040 attcgttctt gatggtttcc tccacagttt ttctccataa tcttgaagag gccaaaacat 2100 tagctttatc caaggaccaa ataggcaatg gtggctcatg ttgtagggcc atgaaagcgg 2160 ccattcttgt gattctttgc acttctggaa cggtgtattg ttcactatcc caagcgacac 2220 catcaccatc gtcttccttt ctcttaccaa agtaaatacc tcccactaat tctctgacaa 2280 caacgaagtc agtaccttta gcaaattgtg gcttgattgg agataagtct aaaagagagt 2340 cggatgcaaa gttacatggt cttaagttgg cgtacaattg aagttcttta cggattttta 2400 gtaaaccttg ttcaggtcta acactacctg taccccattt aggaccaccc acagcaccta 2460 acaaaacggc atcaaccttc ttggaggctt ccagcgcctc atctggaagt gggacacctg 2520 tagcatcgat agcagcacca ccaattaaat gattttcgaa atcgaacttg acattggaac 2580 gaacatcaga aatagcttta agaaccttaa tggcttcggc tgtgatttct tgaccaacgt 2640 ggtcacctgg caaaacgacg atcttcttag gggcagacat tagaatggta tatccttgaa 2700 atatatatat atattgctga aatgtaaaag gtaagaaaag ttagaaagta agacgattgc 2760 taaccaccta ttggaaaaaa caataggtcc ttaaataata ttgtcaactt caagtattgt 2820 gatgcaagca tttagtcatg aacgcttctc tattctatat gaaaagccgg ttccggcctc 2880 tcacctttcc tttttctccc aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc 2940 tgaaattaac aaaaaatttc cagtcatcga atttgattct gtgcgatagc gcccctgtgt 3000 gttctcgtta tgttgaggaa aaaaataatg gttgctaaga gattcgaact cttgcatctt 3060 acgatacctg agtattccca cagttgggga tctcgactct agctagagga tcaattcgta 3120 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 3180 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt 3240 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctggatta 3300 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 3360 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 3420 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 3480 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 3540 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 3600 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 3660 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 3720 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 3780 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 3840 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 3900 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 3960 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 4020 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 4080 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 4140 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 4200 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 4260 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 4320 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 4380 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 4440 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 4500 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 4560 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 4620 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 4680 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 4740 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 4800 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 4860 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 4920 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 4980 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 5040 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 5100 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 5160 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 5220 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 5280 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc 5340 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga 5400 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc 5460 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact 5520 gagagtgcac cataacgcat ttaagcataa acacgcacta tgccgttctt ctcatgtata 5580 tatatataca ggcaacacgc agatataggt gcgacgtgaa cagtgagctg tatgtgcgca 5640 gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg ctttgaagtt cctattccga 5700 agttcctatt ctctagctag aaagtatagg aacttcagag cgcttttgaa aaccaaaagc 5760 gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa 5820 tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta tatatctctg 5880 tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg catctaaact 5940 cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa aaattgtagt 6000 aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt cctatacgta 6060 gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga agaatcatca 6120 acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga atataatcgg 6180 ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt aaacgcggga 6240 agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc ttcttctaac 6300 cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc gcacaaagga 6360 gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat gcatttttgt 6420 agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg ttgcatttct 6480 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt 6540 tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt tcgcgttgca 6600 tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt 6660 gcatttttgt tctacaaaat gaagcacaga tgcttcgtt 6699 <210> 70 <211> 6481 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 70 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc 240 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca 300 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat 360 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc 420 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc 480 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt 540 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg 600 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct 660 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg 720 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct 780 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac 840 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat 900 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc 960 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg 1020 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca 1080 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc 1140 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata 1200 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact 1260 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc 1320 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca 1380 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt 1440 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca 1500 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg 1560 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca 1620 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga 1680 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc 1740 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata 1800 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat 1860 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat 1920 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct 1980 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca 2040 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat 2100 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga 2160 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgccgc atgccggtag 2220 aggtgtggtc aataagagcg acctcatgct atacctgaga aagcaacctg acctacagga 2280 aagagttact caagaataag aattttcgtt ttaaaaccta agagtcactt taaaatttgt 2340 atacacttat tttttttata acttatttaa taataaaaat cataaatcat aagaaattcg 2400 cttatttaga agtgtcaaca acgtatctac caacgatttg acccttttcc atcttttcgt 2460 aaatttctgg caaggtagac aagccgacaa ccttgattgg agacttgacc aaacctctgg 2520 cgaagaagtc caaagcttct gaataagccc tcgtaatata ttttcatgaa gaatttaggt 2580 ccaaaaaaaa gatgggcatt aattctagtc atttaaaaaa ttctatagat cagaggttac 2640 atggccaaga ttgaaactta gaggagtata gttacataaa agaaggcaaa acgatgtata 2700 aatgaaagaa attgagatgg tgcacgatgc acagttgaag tgaacttgcg gggtttttca 2760 gtatctacga ttcatagatc tggaattggg ccggcctggc cactagtcat catgatatcc 2820 tcgagggccg cctgggcctg tcgagagatc tctttttttg ggtttggtgg ggtatcttca 2880 tcatcgaata gatagttata tacatcatcc attgtagtgg tattaaacat ccctgtagtg 2940 attccaaacg cgttatacgc agtttggtcc gtccaaccag gtgacagtgg ttttgaatta 3000 ttaccatcat caattttact agccgtgatt tcattattca tgaagttatc atgaacgtta 3060 gaggaggcaa ttggttgtga aagcgcttga gaatttgttt gagttgttat gaggttcgga 3120 ccgttgctac tgttagtgaa agtgaaggac aatgagctat cagcaatatt cccactttga 3180 ttaaaattgg cggcggtacc caattcgacc tttctcttct tttttggagg ctcgggaatt 3240 aattccgctt tatccatctt tgcaaagctt ggagttgatt gtatgcttgg tatagcttga 3300 aatattgtgc agaaaaagaa acaaggaaga aagggaacga gaacaatgac gaggaaacaa 3360 aagattaata attgcaggtc tatttatact tgatagcaaa gcggcaaact ttttttattt 3420 caaattcaag taactggaag gaaggccgta taccgttgct cattagagag tagtgtgcgt 3480 gaatgaagga aggaaaaagt ttcgtgtgtt cgaagatacc cctcatcagc tctggaacaa 3540 cgacatctgt tggtgctgtc tttgtcgtta attttttcct ttagtgtctt ccatcatttt 3600 ttttgtcatt gcggatatgg tgagacaaca acgggggaga gagaaaagaa aaaaaaagaa 3660 aagaagttgc atgcattcat gcgggcccgg tacccagctt ttgttccctt tagtgagggt 3720 taattccgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 3780 tcacaattcc acacaacata ggagccggaa gcataaagtg taaagcctgg ggtgcctaat 3840 gagtgaggta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 3900 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 3960 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 4020 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 4080 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 4140 tggcgttttt ccataggctc ggcccccctg acgagcatca caaaaatcga cgctcaagtc 4200 agaggtggcg aaacccgaca ggactataaa gataccaggc gttcccccct ggaagctccc 4260 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 4320 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg 4380 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 4440 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 4500 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 4560 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 4620 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 4680 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 4740 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 4800 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 4860 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 4920 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactgc 4980 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 5040 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 5100 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 5160 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 5220 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 5280 aacgatcaag gcgagttaca tgatccccca tgttgtgaaa aaaagcggtt agctccttcg 5340 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 5400 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 5460 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 5520 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 5580 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 5640 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 5700 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 5760 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 5820 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 5880 cccgaaaagt gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa 5940 ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa 6000 tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt 6060 ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag 6120 aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat 6180 ctatttaatc tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa 6240 ttttttaaac ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt 6300 tactttctaa aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa 6360 ttattccatc attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtcctaa 6420 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt 6480 c 6481 <210> 71 <211> 6018 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 71 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc 240 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca 300 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat 360 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc 420 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc 480 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt 540 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg 600 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct 660 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg 720 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct 780 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac 840 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat 900 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc 960 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg 1020 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca 1080 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc 1140 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata 1200 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact 1260 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc 1320 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca 1380 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt 1440 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca 1500 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg 1560 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca 1620 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga 1680 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc 1740 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata 1800 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat 1860 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat 1920 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct 1980 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca 2040 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat 2100 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga 2160 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg 2220 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt 2280 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg 2340 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta 2400 gaagttctcc tcgagggtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa 2460 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt 2520 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa 2580 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 2640 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 2700 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 2760 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 2820 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 2880 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg 2940 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc 3000 cagctggcga aggggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc 3060 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa 3120 ttggagctcc accgcggtgg cggccgctct agaactagtg gatcccccgg gctgcaggaa 3180 ttcgatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccagcttttg 3240 ttccctttag tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt 3300 gtgaaattgt tatccgctca caattccaca caacatagga gccggaagca taaagtgtaa 3360 agcctggggt gcctaatgag tgaggtaact cacattaatt gcgttgcgct cactgcccgc 3420 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 3480 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 3540 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 3600 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 3660 taaaaaggcc gcgttgctgg cgtttttcca taggctcggc ccccctgacg agcatcacaa 3720 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 3780 cccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 3840 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct 3900 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 3960 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 4020 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 4080 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat 4140 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 4200 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 4260 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 4320 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 4380 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 4440 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 4500 catagttgcc tgactgcccg tcgtgtagat aactacgata cgggagggct taccatctgg 4560 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 4620 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 4680 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 4740 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 4800 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa 4860 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 4920 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 4980 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 5040 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 5100 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 5160 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 5220 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 5280 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 5340 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 5400 ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa 5460 aataattata atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa 5520 agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg 5580 ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt 5640 tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc 5700 gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag 5760 tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact 5820 cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca 5880 cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg 5940 caagcgatcc gtcctaagaa accattatta tcatgacatt aacctataaa aataggcgta 6000 tcacgaggcc ctttcgtc 6018 <210> 72 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 72 cgccgcatgc attcatgcag gcc 23 <210> 73 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 73 tgcatgaatg catgcgg 17 <210> 74 <211> 5021 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 74 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc 240 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca 300 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat 360 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc 420 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc 480 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt 540 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg 600 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct 660 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg 720 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct 780 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac 840 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat 900 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc 960 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg 1020 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca 1080 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc 1140 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata 1200 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact 1260 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc 1320 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca 1380 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt 1440 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca 1500 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg 1560 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca 1620 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga 1680 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc 1740 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata 1800 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat 1860 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat 1920 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct 1980 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca 2040 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat 2100 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga 2160 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgccgc atgcattcat 2220 gcaggcccgg tacccagctt ttgttccctt tagtgagggt taattccgag cttggcgtaa 2280 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 2340 ggagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta 2400 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 2460 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 2520 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 2580 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 2640 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 2700 ggcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 2760 ggactataaa gataccaggc gttcccccct ggaagctccc tcgtgcgctc tcctgttccg 2820 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 2880 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 2940 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 3000 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 3060 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 3120 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 3180 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 3240 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 3300 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 3360 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 3420 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 3480 gcgatctgtc tatttcgttc atccatagtt gcctgactgc ccgtcgtgta gataactacg 3540 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 3600 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 3660 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 3720 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 3780 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 3840 tgatccccca tgttgtgaaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 3900 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 3960 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 4020 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 4080 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 4140 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 4200 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 4260 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 4320 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 4380 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg 4440 tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata taaatatata 4500 aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga 4560 tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct cttcctgctc 4620 tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac acgaaaatcc 4680 tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc tgcttttctt 4740 gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac ctttgtttat 4800 ttttttttct tcattccgta actcttctac cttctttatt tactttctaa aatccaaata 4860 caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc attaaaagat 4920 acgaggcgcg tgtaagttac aggcaagcga tccgtcctaa gaaaccatta ttatcatgac 4980 attaacctat aaaaataggc gtatcacgag gccctttcgt c 5021 <210> 75 <211> 6408 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <400> 75 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc 240 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca 300 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat 360 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc 420 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc 480 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt 540 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg 600 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct 660 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg 720 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct 780 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac 840 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat 900 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc 960 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg 1020 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca 1080 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc 1140 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata 1200 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact 1260 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc 1320 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca 1380 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt 1440 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca 1500 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg 1560 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca 1620 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga 1680 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc 1740 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata 1800 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat 1860 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat 1920 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct 1980 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca 2040 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat 2100 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga 2160 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgccgc atgccggtag 2220 aggtgtggtc aataagagcg acctcatgct atacctgaga aagcaacctg acctacagga 2280 aagagttact caagaataag aattttcgtt ttaaaaccta agagtcactt taaaatttgt 2340 atacacttat tttttttata acttatttaa taataaaaat cataaatcat aagaaattcg 2400 cttatttaga agtgtcaaca acgtatctac caacgatttg acccttttcc atcttttcgt 2460 aaatttctgg caaggtagac aagccgacaa ccttgattgg agacttgacc aaacctctgg 2520 cgaagaagtc caaagcttct gaataagccc tcgtaatata ttttcatgaa gaatttaggt 2580 ccaaaaaaaa gatgggcatt aattctagtc atttaaaaaa ttctatagat cagaggttac 2640 atggccaaga ttgaaactta gaggagtata gttacataaa agaaggcaaa acgatgtata 2700 aatgaaagaa attgagatgg tgcacgatgc acagttgaag tgaacttgcg gggtttttca 2760 gtatctacga ttcatagatc tggaattggg ccggcctggc cactagtcat catgatatcc 2820 tcgagggccg cctgggcctg tcgagagatc tctttttttg ggtttggtgg ggtatcttca 2880 tcatcgaata gatagttata tacatcatcc attgtagtgg tattaaacat ccctgtagtg 2940 attccaaacg cgttatacgc agtttggtcc gtccaaccag gtgacagtgg ttttgaatta 3000 ttaccatcat caattttact agccgtgatt tcattattca tgaagttatc atgaacgtta 3060 gaggaggcaa ttggttgtga aagcgcttga gaatttgttt gagttgttat gaggttcgga 3120 ccgttgctac tgttagtgaa agtgaaggac aatgagctat cagcaatatt cccactttga 3180 ttaaaattgg cggcggtacc caattcgacc tttctcttct tttttggagg ctcgggaatt 3240 aattccgctt tatccatctt tgcagcggcc gcttgcaaaa gcctaggcct ccaaaaaagc 3300 ctcctcacta cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa 3360 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga 3420 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac 3480 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca 3540 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca 3600 cattccacag ggcccggtac ccagcttttg ttccctttag tgagggttaa ttccgagctt 3660 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 3720 caacatagga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgaggtaact 3780 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 3840 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 3900 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 3960 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 4020 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 4080 taggctcggc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 4140 cccgacagga ctataaagat accaggcgtt cccccctgga agctccctcg tgcgctctcc 4200 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 4260 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 4320 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 4380 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 4440 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 4500 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 4560 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 4620 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 4680 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 4740 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 4800 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 4860 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactgcccg tcgtgtagat 4920 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 4980 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 5040 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 5100 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 5160 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 5220 agttacatga tcccccatgt tgtgaaaaaa agcggttagc tccttcggtc ctccgatcgt 5280 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 5340 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 5400 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 5460 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 5520 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 5580 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 5640 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 5700 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 5760 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 5820 acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa 5880 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt 5940 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt 6000 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg 6060 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc 6120 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt 6180 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat 6240 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt 6300 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa accattatta 6360 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc 6408 <210> 76 <211> 6308 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <400> 76 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc 240 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca 300 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat 360 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc 420 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc 480 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt 540 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg 600 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct 660 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg 720 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct 780 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac 840 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat 900 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc 960 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg 1020 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca 1080 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc 1140 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata 1200 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact 1260 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc 1320 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca 1380 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt 1440 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca 1500 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg 1560 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca 1620 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga 1680 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc 1740 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata 1800 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat 1860 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat 1920 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct 1980 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca 2040 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat 2100 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga 2160 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgccgc atgccggtag 2220 aggtgtggtc aataagagcg acctcatgct atacctgaga aagcaacctg acctacagga 2280 aagagttact caagaataag aattttcgtt ttaaaaccta agagtcactt taaaatttgt 2340 atacacttat tttttttata acttatttaa taataaaaat cataaatcat aagaaattcg 2400 cttatttaga agtgtcaaca acgtatctac caacgatttg acccttttcc atcttttcgt 2460 aaatttctgg caaggtagac aagccgacaa ccttgattgg agacttgacc aaacctctgg 2520 cgaagaagtc caaagcttct gaataagccc tcgtaatata ttttcatgaa gaatttaggt 2580 ccaaaaaaaa gatgggcatt aattctagtc atttaaaaaa ttctatagat cagaggttac 2640 atggccaaga ttgaaactta gaggagtata gttacataaa agaaggcaaa acgatgtata 2700 aatgaaagaa attgagatgg tgcacgatgc acagttgaag tgaacttgcg gggtttttca 2760 gtatctacga ttcatagatc tggaattggg ccggcctggc cactagtcat catgatatcc 2820 tcgagggccg cctgggcctg tcgagagatc tctttttttg ggtttggtgg ggtatcttca 2880 tcatcgaata gatagttata tacatcatcc attgtagtgg tattaaacat ccctgtagtg 2940 attccaaacg cgttatacgc agtttggtcc gtccaaccag gtgacagtgg ttttgaatta 3000 ttaccatcat caattttact agccgtgatt tcattattca tgaagttatc atgaacgtta 3060 gaggaggcaa ttggttgtga aagcgcttga gaatttgttt gagttgttat gaggttcgga 3120 ccgttgctac tgttagtgaa agtgaaggac aatgagctat cagcaatatt cccactttga 3180 ttaaaattgg cggcggtacc caattcgacc tttctcttct tttttggagg ctcgggaatt 3240 aattccgctt tatccatctt tgcagcggcc gcagccatgg ggcggagaat gggcggaact 3300 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt 3360 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt 3420 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt 3480 tccacaccct aactgacaca cattccacag ggcccggtac ccagcttttg ttccctttag 3540 tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 3600 tatccgctca caattccaca caacatagga gccggaagca taaagtgtaa agcctggggt 3660 gcctaatgag tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 3720 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 3780 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 3840 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 3900 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 3960 gcgttgctgg cgtttttcca taggctcggc ccccctgacg agcatcacaa aaatcgacgc 4020 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt cccccctgga 4080 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4140 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 4200 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4260 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4320 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4380 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 4440 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4500 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 4560 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 4620 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 4680 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 4740 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 4800 tgactgcccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 4860 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 4920 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 4980 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 5040 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 5100 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa agcggttagc 5160 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 5220 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 5280 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 5340 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 5400 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 5460 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 5520 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 5580 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 5640 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 5700 acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata 5760 atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa 5820 gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata 5880 ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc 5940 tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa 6000 tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt 6060 gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt 6120 ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat 6180 tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc 6240 gtcctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc 6300 ctttcgtc 6308 <210> 77 <211> 7730 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <220> <221> misc_feature <222> (4662) <223> n is a, c, g, or t <220> <221> misc_feature <222> (6401) <223> n is a, c, g, or t <400> 77 tctctggcta actagagaac ccactgctta ctggcttatc gaaattttaa ttaacgttgg 60 caccatgctg ctgctgctgc tgctgctggg cctgaggcta cagctctccc tgggcatcat 120 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc 180 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga 240 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa 300 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac 360 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg 420 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa 480 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc 540 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca 600 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg 660 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg 720 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag 780 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca 840 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt 900 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc 960 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa 1020 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag 1080 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg 1140 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt 1200 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc 1260 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa 1320 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca 1380 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg 1440 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat 1500 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac 1560 caccgacgcc gcgcacccgg gttactctag agtcggggcg gccggctagg tttaaacact 1620 agaaataatt cttactgtca tgccaagtaa gatgcttttc tgtgctgcaa tagcaggcat 1680 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaactagacc cagctttctt 1740 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac 1800 tatcagtcaa aataaaatca ttatttgcca tccaggtcga gtgtggaatg tgtgtcagtt 1860 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 1920 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 1980 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 2040 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 2100 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg 2160 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca 2220 aagatccacc ggagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg 2280 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc 2340 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc 2400 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg 2460 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc 2520 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg 2580 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc 2640 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg 2700 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc 2760 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga 2820 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg 2880 cacgacccca tgcatcggta cctagagtcg gggcggccgg ccgcttcgag cagacatgat 2940 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat 3000 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt 3060 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt 3120 ttaaagcaag taaaacctct acaaatgtgg taaaatcgct gcagctctgg cccgtgtctc 3180 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt 3240 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga 3300 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata 3360 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt 3420 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac 3480 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg 3540 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag 3600 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc 3660 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg 3720 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg 3780 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt 3840 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa 3900 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc 3960 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg 4020 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct 4080 aatcagaatt ggttaattgg ttgtaacatt attcagattg ggccccgttc cactgagcgt 4140 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 4200 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 4260 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 4320 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 4380 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 4440 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 4500 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 4560 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 4620 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gngaaacgcc tggtatcttt 4680 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 4740 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 4800 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 4860 ttaccgctag catggatctc ggggacgtct aactactaag cgagagtagg gaactgccag 4920 gcatcaaata aaacgaaagg ctcagtcgga agactgggcc tttcgtttta tctgttgttt 4980 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgtgaa 5040 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaactaa 5100 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttc ctgttagtta 5160 gttacttaag ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt 5220 gcaacaaatt gataagcaat gcttttttat aatgccaact ttgtacaaaa aagcaggctt 5280 cgaaggagat agaaccagat cttggaattc tgcagatatc gaaatttggg aggggagcca 5340 tcaaagaagc ctgggagcag cagttccagg gaaaaaggag aatgtgatgg ccagagagcc 5400 aaaagaaaaa gtagttgaag gagtgctcag cactaggcat ctgaactgaa tgctgtggca 5460 ggctcactgg ccacaaacaa tagggagctg gtggaggcct tgacgaggac catttcaaca 5520 aactggtggg cttaaaatcc ggaagaaaca gttgaacaaa tcattttgac gccttttata 5580 aaccacacaa gcttattcca aacccgttac tggcctaact gatttaagtc cctttcccat 5640 ctgatcctca gagattctaa gggacttagc ctatccatga ctcttcgtcc tgcttctcac 5700 ctcccatgat tgccctaacg atgtgaaagt gctttcaaac aaagatgccc aagaaagaag 5760 gtaggcaaat gtgcaagcat tagtttgtag tacgctatta ctgtatttca ccttgcactc 5820 tctagtttcc ttcgtgctcc ctcaatatcc aactcttaat aaattcatgg ctcccggtga 5880 gcattcatca attctcattc cacgccttta gcccttcccg ttcccgccca actctcgctc 5940 cctcccctgg ccaaatctct aacctgcaag gctaattccg aattccaaat cggaagcaag 6000 agggcggggc cccgtgagag gcgatggatt gctccagtcc gttcccgacg cactgtgcgc 6060 atgcgctggt cctccgcgga ccgttcgtgc tgcccgccta gaaagggtga agtggttgtt 6120 tccgtgacgg actgagtacg ggtgcctgtc aggctcttgc ggaagtccat gcgccattgg 6180 gagggcctcg gccgcggctc tgtgcccttg ctgctgaggg ccacttcctg ggtcattcct 6240 ggaccgggag ccgggctggg gctcacacgg gggctcccgc gtggccgtct cggcgcctgc 6300 gtgacctccc cgccggcggg ctcgagccca agcttggtac cgagctcgga tccagccacc 6360 atgggagtca aagttctgtt tgccctgatc tgcatcgctg nggccgaggc caagcccacc 6420 gagaacaacg aagacttcaa catcgtggcc gtggccagca acttcgcgac cacggatctc 6480 gatgctgacc gcgggaagtt gcccggcaag aagctgccgc tggaggtgct caaagagctg 6540 gaagccaatg cccggaaagc tggctgcacc aggggctgtc tgatctgcct gtcccacatc 6600 aagtgcacgc ccaagatgaa gaagttcatc ccaggacgct gccacaccta cgaaggcgac 6660 aaagagtccg cacagggcgg cataggcgag gcgatcgtcg acattcctga gattcctggg 6720 ttcaaggact tggagcccct ggagcagttc atcgcacagg tcgatctgtg tgtggactgc 6780 acaactggct gcctcaaagg gcttgccaac gtgcagtgtt ctgacctgct caagaagtgg 6840 ctgccgcaac gctgtgcgac ctttgccagc aagatccagg gccaggtgga caagatcaag 6900 ggggccggtg gtgactaagc ggccgcttcg agcagacatg ataagataca ttgatgagtt 6960 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 7020 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat 7080 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct 7140 ctacaaatgt ggtacaaccg gtctagttat taatagtaat caattacggg gtcattagtt 7200 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga 7260 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca 7320 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 7380 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 7440 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 7500 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 7560 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 7620 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 7680 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 7730 <210> 78 <211> 6083 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 78 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 60 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 120 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 180 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 240 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 300 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 360 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 420 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 480 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 540 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 600 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 660 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 720 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 780 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 840 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 900 gtttaaacgg gccctctaga gatatcatgg atgctaagtc cctgacagcg tggagccgca 960 cactggttac cttcaaagat gttttcgtgg atttcacccg cgaagagtgg aaactgctgg 1020 ataccgcaca gcagattgtg tatcgcaacg ttatgctgga aaactacaag aatctggtta 1080 gcctgggcta tcagctgaca aaacccgacg tcatcctgcg tctggaaaag ggtgaagagc 1140 cgtggctggt tgaacgggag attcaccagg agacacatcc tgattctgaa actgcctttg 1200 agatcaaaag ctccgtcagt ccgaaaaaga aacgtaaagt ggggctcgag cccggggaaa 1260 agccatataa atgccccgag tgcggcaaat cattcagcca aagtagcaac ttagtaagac 1320 accagcgcac ccataccggg gaaaagccat ataaatgccc cgagtgcggc aaatcattca 1380 gccaaagtag caacttagta agacaccagc gcacccatac cggggaaaag ccatataaat 1440 gccccgagtg cggcaaatca ttcagccaaa gtagcaactt agtaagacac cagcgcaccc 1500 ataccggtga gcagaaactc atctctgaag aagatctgga acaaaagttg atttcagaag 1560 aagatctgga acagaagctc atctctgagg aagatctgta agcggccgcg aattccacca 1620 cactggacta gtggatccga gctcggtacc aagcttaagt ttaaaccgct gatcagcctc 1680 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac 1740 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg 1800 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga 1860 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga 1920 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc 1980 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 2040 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 2100 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 2160 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 2220 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 2280 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 2340 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt 2400 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 2460 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2520 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 2580 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 2640 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt 2700 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct 2760 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt 2820 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 2880 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 2940 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg 3000 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac 3060 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 3120 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 3180 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 3240 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 3300 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat 3360 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 3420 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 3480 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 3540 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt 3600 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg 3660 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc 3720 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata 3780 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 3840 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga 3900 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 3960 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 4020 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 4080 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 4140 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 4200 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 4260 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 4320 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 4380 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 4440 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 4500 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 4560 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 4620 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 4680 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 4740 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 4800 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 4860 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 4920 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 4980 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 5040 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 5100 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 5160 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 5220 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 5280 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 5340 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 5400 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 5460 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 5520 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 5580 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 5640 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 5700 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 5760 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 5820 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 5880 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 5940 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 6000 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6060 cccgaaaagt gccacctgac gtc 6083 <210> 79 <211> 5916 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 79 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 60 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 120 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 180 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 240 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 300 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 360 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 420 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 480 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 540 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 600 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 660 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 720 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 780 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 840 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 900 gtttaaacgg gccctctaga gatatcatgc cgaaaaagaa acgtaaagtg gggctcgagc 960 ccggggaaaa gccatataaa tgccccgagt gcggcaaatc attcagccaa agtagcaact 1020 tagtaagaca ccagcgcacc cataccgggg aaaagccata taaatgcccc gagtgcggca 1080 aatcattcag ccaaagtagc aacttagtaa gacaccagcg cacccatacc ggggaaaagc 1140 catataaatg ccccgagtgc ggcaaatcat tcagccaaag tagcaactta gtaagacacc 1200 agcgcaccca taccggtggc ggcagcggcg gcagcgaatt ccgcacactg gttaccttca 1260 aagatgtttt cgtggatttc acccgcgaag agtggaaact gctggatacc gcacagcaga 1320 ttgtgtatcg caacgttatg ctggaaaact acaagaatct ggttagcctg ggctatggat 1380 ccgagcagaa actcatctct gaagaagatc tggaacaaaa gttgatttca gaagaagatc 1440 tggaacagaa gctcatctct gaggaagatc tgtaagcggc cgcaagctta agtttaaacc 1500 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg 1560 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa 1620 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca 1680 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg 1740 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg 1800 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 1860 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 1920 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc 1980 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 2040 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 2100 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga 2160 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct 2220 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 2280 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 2340 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 2400 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 2460 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta 2520 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc 2580 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg 2640 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 2700 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 2760 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg 2820 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 2880 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 2940 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 3000 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 3060 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 3120 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt 3180 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 3240 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 3300 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 3360 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc 3420 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 3480 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 3540 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 3600 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 3660 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 3720 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 3780 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 3840 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 3900 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 3960 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 4020 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 4080 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 4140 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 4200 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 4260 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 4320 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 4380 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 4440 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 4500 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 4560 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 4620 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 4680 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 4740 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 4800 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 4860 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 4920 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 4980 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 5040 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 5100 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 5160 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 5220 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 5280 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 5340 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 5400 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 5460 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 5520 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 5580 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 5640 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 5700 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 5760 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 5820 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 5880 ccgcgcacat ttccccgaaa agtgccacct gacgtc 5916 <210> 80 <211> 5897 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 80 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 60 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 120 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 180 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 240 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 300 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 360 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 420 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 480 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 540 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 600 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 660 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 720 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 780 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 840 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 900 gtttaaacgg gccctctaga gatatcatgg cggcggcggt tcggatgaac atccagatgc 960 tgctggaggc ggccgactat ctggagcggc gggagagaga agctgaacat ggttatgcct 1020 ccatgttacc atacccgaaa aagaaacgta aagtggggct cgagcccggg gaaaagccat 1080 ataaatgccc cgagtgcggc aaatcattca gccaaagtag caacttagta agacaccagc 1140 gcacccatac cggggaaaag ccatataaat gccccgagtg cggcaaatca ttcagccaaa 1200 gtagcaactt agtaagacac cagcgcaccc ataccgggga aaagccatat aaatgccccg 1260 agtgcggcaa atcattcagc caaagtagca acttagtaag acaccagcgc acccataccg 1320 gtgagcagaa actcatctct gaagaagatc tggaacaaaa gttgatttca gaagaagatc 1380 tggaacagaa gctcatctct gaggaagatc tgtaagcggc cgcgaattcc accacactgg 1440 actagtggat ccgagctcgg taccaagctt aagtttaaac cgctgatcag cctcgactgt 1500 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 1560 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 1620 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 1680 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 1740 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 1800 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 1860 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 1920 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 1980 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 2040 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 2100 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 2160 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 2220 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 2280 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 2340 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 2400 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 2460 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 2520 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 2580 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 2640 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 2700 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 2760 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 2820 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 2880 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 2940 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 3000 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 3060 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 3120 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 3180 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 3240 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 3300 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 3360 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 3420 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 3480 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 3540 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 3600 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 3660 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 3720 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 3780 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 3840 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 3900 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 3960 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 4020 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 4080 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 4140 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 4200 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 4260 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 4320 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 4380 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 4440 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 4500 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 4560 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 4620 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 4680 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 4740 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 4800 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 4860 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 4920 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 4980 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 5040 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 5100 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 5160 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 5220 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 5280 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 5340 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 5400 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 5460 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 5520 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 5580 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 5640 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 5700 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 5760 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 5820 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 5880 aagtgccacc tgacgtc 5897 <210> 81 <211> 6198 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 81 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 60 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 120 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 180 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 240 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 300 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 360 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 420 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 480 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 540 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 600 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 660 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 720 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 780 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 840 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 900 gtttaaacgg gccctctaga gatatcatgc cgaaaaagaa acgtaaagtg gggctcgagc 960 ccggggaaaa gccctacaag tgccctgagt gtgggaagtc cttttcttca agacgcacgt 1020 gccgcgctca ccagcggaca cataccgggg agaagcccta taaatgtcca gaatgtggaa 1080 agtcctttag cacgtcaggg aacttagtaa gacaccagcg aactcatacc ggggagaagc 1140 catataaatg tcccgagtgt ggcaagtcct tttctagatc agataattta gtaagacatc 1200 agagaacgca caccggggaa aagccctaca agtgcccgga atgcggcaag tcttttagca 1260 ccagcggaca tttagtaaga caccagagaa cccacaccgg ggaaaaaccc tataaatgcc 1320 ccgagtgtgg taagtcattc tctcaaagcg gggatttaag aagacaccag agaacccaca 1380 ccggggaaaa accgtataaa tgtcctgagt gcggtaagtc tttttccgac tgtagagact 1440 tagcgagaca ccaacgtact cataccggtg gcggcagcgg cggcagcgaa ttcgggcgcg 1500 ccgacgcgct ggacgatttc gatctcgaca tgctgggttc tgatgccctc gatgactttg 1560 acctggatat gttgggaagc gacgcattgg atgactttga tctggacatg ctcggctccg 1620 atgctctgga cgatttcgat ctcgatatgt taattaacgg atccgagcag aaactcatct 1680 ctgaagaaga tctggaacaa aagttgattt cagaagaaga tctggaacag aagctcatct 1740 ctgaggaaga tctgtaagcg gccgcaagct taagtttaaa ccgctgatca gcctcgactg 1800 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 1860 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 1920 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 1980 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa 2040 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg 2100 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 2160 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 2220 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 2280 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 2340 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 2400 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 2460 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt 2520 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 2580 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 2640 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 2700 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 2760 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg 2820 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca 2880 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 2940 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 3000 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 3060 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 3120 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 3180 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 3240 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 3300 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 3360 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 3420 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 3480 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 3540 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 3600 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 3660 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa 3720 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 3780 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 3840 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 3900 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 3960 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 4020 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 4080 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 4140 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 4200 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 4260 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 4320 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 4380 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 4440 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 4500 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 4560 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 4620 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 4680 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 4740 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 4800 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 4860 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 4920 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 4980 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 5040 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 5100 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 5160 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 5220 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 5280 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 5340 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 5400 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 5460 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 5520 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 5580 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 5640 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 5700 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 5760 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 5820 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 5880 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 5940 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 6000 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 6060 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 6120 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 6180 aaagtgccac ctgacgtc 6198 <210> 82 <211> 5185 <212> DNA <213> Artificial Sequence <220> <223> Synthetic construct <400> 82 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc 3180 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc 3240 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3300 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3360 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct 3420 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc 3480 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt 3540 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca 3600 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat 3660 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg 3720 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg 3780 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca 3840 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg 3900 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa 3960 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa 4020 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca 4080 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt 4140 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt 4200 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg 4260 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac 4320 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac 4380 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg 4440 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg 4500 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt 4560 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 4620 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 4680 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat 4740 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta 4800 gaaataattt tgtttaactt taagaaggag atatacatat gcaccaccac caccaccacg 4860 gctatggccg caaaaaacgc cgccagcgcc gccgcggcta tccgtatgat gtgccggatt 4920 atgccccatg ggatatcggg gatccgaatt ctgtacaggc cttggcgcgc ctgcaggcga 4980 gctccgtcga caagcttgcg gccgcactcg agcaccacca ccaccaccac caccactaat 5040 tgattaatac ctaggctgct aaacaaagcc cgaaaggaag ctgagttggc tgctgccacc 5100 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg 5160 ctgaaaggag gaactatatc cggat 5185 <210> 83 <211> 5956 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <220> <221> misc_feature <222> (4978) <223> n is a, c, g, or t <400> 83 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc 3180 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc 3240 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3300 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3360 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct 3420 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc 3480 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt 3540 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca 3600 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat 3660 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg 3720 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg 3780 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca 3840 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg 3900 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa 3960 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa 4020 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca 4080 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt 4140 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt 4200 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg 4260 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac 4320 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac 4380 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg 4440 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg 4500 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt 4560 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 4620 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 4680 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat 4740 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta 4800 gaaataattt tgtttaactt taagaaggag atatacatat gcaccaccac caccaccacg 4860 gctatggccg caaaaaacgc cgccagcgcc gccgcggcta tccgtatgat gtgccggatt 4920 atgccccatg ggatatcatg ccgaaaaaga aacgtaaagt ggggctcgag cccggggnga 4980 agccctataa atgccctgaa tgcgggaaat ctttctcttc taagaaggca ctcacagaac 5040 accagcggac acacaccggg gaaaaaccgt acaagtgtcc tgagtgcggg aagagtttct 5100 ccgatccggg ccacttagta agacatcaga ggacacatac cggggagaag ccatataaat 5160 gtcccgagtg tggcaagtcc ttttctagat cagataattt agtaagacat cagagaacgc 5220 acaccgggga gaagccatat aaatgtcccg agtgtggcaa gtccttttct agatcagata 5280 atttagtaag acatcagaga acgcacaccg gggaaaagcc atataaatgc cccgagtgcg 5340 gcaaatcatt cagccaaagt agcaacttag taagacacca gcgcacccat accggggaaa 5400 aaccgtacaa gtgtcctgag tgcgggaaga gtttctccga tccgggccac ttagtaagac 5460 atcagaggac acataccggt ggcggcagcg gcggcagcga attcgggcgc gccgacgcgc 5520 tggacgattt cgatctcgac atgctgggtt ctgatgccct cgatgacttt gacctggata 5580 tgttgggaag cgacgcattg gatgactttg atctggacat gctcggctcc gatgctctgg 5640 acgatttcga tctcgatatg ttaattaacg gatccgagca gaaactcatc tctgaagaag 5700 atctggaaca aaagttgatt tcagaagaag atctggaaca gaagctcatc tctgaggaag 5760 atctgtaagc ggccgcactc gagcaccacc accaccacca ccaccactaa ttgattaata 5820 cctaggctgc taaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 5880 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 5940 ggaactatat ccggat 5956 <210> 84 <211> 5956 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <400> 84 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc 3180 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc 3240 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3300 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3360 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct 3420 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc 3480 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt 3540 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca 3600 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat 3660 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg 3720 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg 3780 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca 3840 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg 3900 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa 3960 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa 4020 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca 4080 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt 4140 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt 4200 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg 4260 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac 4320 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac 4380 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg 4440 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg 4500 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt 4560 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 4620 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 4680 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat 4740 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta 4800 gaaataattt tgtttaactt taagaaggag atatacatat gcaccaccac caccaccacg 4860 gctatggccg caaaaaacgc cgccagcgcc gccgcggcta tccgtatgat gtgccggatt 4920 atgccccatg ggatatcatg ccgaaaaaga aacgtaaagt ggggctcgag cccggggaga 4980 aaccatacaa atgccccgag tgtggaaagt catttagtga tccaggcgca ttagtaagac 5040 atcagcggac acataccggg gagaagccat ataaatgtcc cgagtgtggc aagtcctttt 5100 ctagatcaga taatttagta agacatcaga gaacgcacac cggggagaag ccctacaagt 5160 gtccagaatg cggaaagagt ttctccagaa gtgacaaatt agtaagacac cagagaaccc 5220 ataccgggga aaaaccgtac aagtgtcctg agtgcgggaa gagtttctcc gatccgggcc 5280 acttagtaag acatcagagg acacataccg gggaaaaacc gtataaatgt cctgagtgcg 5340 gtaagtcttt ttccgactgt agagacttag cgagacacca acgtactcat accggggaga 5400 aaccatacaa atgtcccgaa tgtggcaaga gtttcagcag taaaaagcat ctcgctgagc 5460 atcagagaac tcacaccggt ggcggcagcg gcggcagcga attcgggcgc gccgacgcgc 5520 tggacgattt cgatctcgac atgctgggtt ctgatgccct cgatgacttt gacctggata 5580 tgttgggaag cgacgcattg gatgactttg atctggacat gctcggctcc gatgctctgg 5640 acgatttcga tctcgatatg ttaattaacg gatccgagca gaaactcatc tctgaagaag 5700 atctggaaca aaagttgatt tcagaagaag atctggaaca gaagctcatc tctgaggaag 5760 atctgtaagc ggccgcactc gagcaccacc accaccacca ccaccactaa ttgattaata 5820 cctaggctgc taaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 5880 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 5940 ggaactatat ccggat 5956 <210> 85 <211> 5956 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <400> 85 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc 3180 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc 3240 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3300 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3360 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct 3420 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc 3480 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt 3540 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca 3600 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat 3660 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg 3720 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg 3780 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca 3840 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg 3900 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa 3960 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa 4020 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca 4080 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt 4140 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt 4200 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg 4260 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac 4320 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac 4380 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg 4440 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg 4500 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt 4560 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 4620 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 4680 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat 4740 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta 4800 gaaataattt tgtttaactt taagaaggag atatacatat gcaccaccac caccaccacg 4860 gctatggccg caaaaaacgc cgccagcgcc gccgcggcta tccgtatgat gtgccggatt 4920 atgccccatg ggatatcatg ccgaaaaaga aacgtaaagt ggggctcgag cccggggaga 4980 agccgtacaa gtgccctgaa tgtggtaagt cattttcgag aagtgatgaa ttagtaagac 5040 accagcggac tcataccggg gagaagccgt acaagtgccc tgaatgtggt aagtcatttt 5100 cgagaagtga tgaattagta agacaccagc ggactcatac cggggagaag ccctataaat 5160 gtccagaatg tggaaagtcc tttagcacgt cagggaactt agtaagacac cagcgaactc 5220 ataccgggga aaagccttac aaatgccccg aatgtgggaa gagtttcagc cggtctgata 5280 agctgaccga acaccagaga actcataccg gggagaagcc ctataaatgc cctgaatgtg 5340 gcaagagctt cagtactagc gggaatctca ctgaacatca gcgaactcat accggggaaa 5400 aaccttacaa gtgccctgag tgcggcaaga gcttctctca atcaagttca ttagtaagac 5460 accagaggac tcataccggt ggcggcagcg gcggcagcga attcgggcgc gccgacgcgc 5520 tggacgattt cgatctcgac atgctgggtt ctgatgccct cgatgacttt gacctggata 5580 tgttgggaag cgacgcattg gatgactttg atctggacat gctcggctcc gatgctctgg 5640 acgatttcga tctcgatatg ttaattaacg gatccgagca gaaactcatc tctgaagaag 5700 atctggaaca aaagttgatt tcagaagaag atctggaaca gaagctcatc tctgaggaag 5760 atctgtaagc ggccgcactc gagcaccacc accaccacca ccaccactaa ttgattaata 5820 cctaggctgc taaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 5880 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 5940 ggaactatat ccggat 5956 <210> 86 <211> 5956 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <400> 86 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc 3180 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc 3240 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3300 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3360 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct 3420 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc 3480 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt 3540 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca 3600 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat 3660 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg 3720 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg 3780 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca 3840 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg 3900 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa 3960 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa 4020 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca 4080 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt 4140 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt 4200 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg 4260 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac 4320 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac 4380 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg 4440 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg 4500 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt 4560 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 4620 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 4680 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat 4740 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta 4800 gaaataattt tgtttaactt taagaaggag atatacatat gcaccaccac caccaccacg 4860 gctatggccg caaaaaacgc cgccagcgcc gccgcggcta tccgtatgat gtgccggatt 4920 atgccccatg ggatatcatg ccgaaaaaga aacgtaaagt ggggctcgag cccggggaga 4980 aaccttataa atgcccagaa tgcgggaaat cgttcagtca aagagcacat ttagaaagac 5040 atcaacggac ccacaccggg gaaaagccat ataaatgccc cgagtgcggc aaatcattca 5100 gccaaagtag caacttagta agacaccagc gcacccatac cggggaaaag ccctacaagt 5160 gtcctgagtg cggaaagtct ttctccacta gcggttcatt agtaagacac cagaggacac 5220 acaccgggga aaaaccttac aagtgccctg agtgcggcaa gagcttctct caatcaagtt 5280 cattagtaag acaccagagg actcataccg gggagaagcc atacaaatgc cctgagtgtg 5340 gaaagtcatt tagccagcga gctaatctgc gggcccacca gcggacccac accggggaaa 5400 agccatataa atgccccgag tgcggcaaat cattcagcca aagtagcaac ttagtaagac 5460 accagcgcac ccataccggt ggcggcagcg gcggcagcga attcgggcgc gccgacgcgc 5520 tggacgattt cgatctcgac atgctgggtt ctgatgccct cgatgacttt gacctggata 5580 tgttgggaag cgacgcattg gatgactttg atctggacat gctcggctcc gatgctctgg 5640 acgatttcga tctcgatatg ttaattaacg gatccgagca gaaactcatc tctgaagaag 5700 atctggaaca aaagttgatt tcagaagaag atctggaaca gaagctcatc tctgaggaag 5760 atctgtaagc ggccgcactc gagcaccacc accaccacca ccaccactaa ttgattaata 5820 cctaggctgc taaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 5880 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 5940 ggaactatat ccggat 5956 <210> 87 <211> 5956 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <400> 87 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc 3180 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc 3240 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3300 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3360 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct 3420 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc 3480 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt 3540 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca 3600 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat 3660 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg 3720 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg 3780 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca 3840 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg 3900 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa 3960 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa 4020 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca 4080 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt 4140 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt 4200 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg 4260 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac 4320 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac 4380 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg 4440 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg 4500 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt 4560 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 4620 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 4680 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat 4740 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta 4800 gaaataattt tgtttaactt taagaaggag atatacatat gcaccaccac caccaccacg 4860 gctatggccg caaaaaacgc cgccagcgcc gccgcggcta tccgtatgat gtgccggatt 4920 atgccccatg ggatatcatg ccgaaaaaga aacgtaaagt ggggctcgag cccggggaga 4980 aaccatacaa atgtcccgaa tgtggcaaga gtttcagcag taaaaagcat ctcgctgagc 5040 atcagagaac tcacaccggg gaaaaacctt acaagtgccc tgagtgcggc aagagcttct 5100 ctcaatcaag ttcattagta agacaccaga ggactcatac cggggaaaaa ccgtacaagt 5160 gtcctgagtg cgggaagagt ttctccgatc cgggccactt agtaagacat cagaggacac 5220 ataccgggga gaaaccttat aaatgcccag aatgcgggaa atcgttcagt caaagagcac 5280 atttagaaag acatcaacgg acccacaccg gggaaaagcc ctacaagtgt cctgagtgcg 5340 gaaagtcttt ctccactagc ggttcattag taagacacca gaggacacac accggggaaa 5400 aaccttacaa gtgccctgag tgcggcaaga gcttctctca atcaagttca ttagtaagac 5460 accagaggac tcataccggt ggcggcagcg gcggcagcga attcgggcgc gccgacgcgc 5520 tggacgattt cgatctcgac atgctgggtt ctgatgccct cgatgacttt gacctggata 5580 tgttgggaag cgacgcattg gatgactttg atctggacat gctcggctcc gatgctctgg 5640 acgatttcga tctcgatatg ttaattaacg gatccgagca gaaactcatc tctgaagaag 5700 atctggaaca aaagttgatt tcagaagaag atctggaaca gaagctcatc tctgaggaag 5760 atctgtaagc ggccgcactc gagcaccacc accaccacca ccaccactaa ttgattaata 5820 cctaggctgc taaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 5880 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 5940 ggaactatat ccggat 5956 <210> 88 <211> 5956 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <400> 88 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc 3180 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc 3240 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3300 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3360 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct 3420 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc 3480 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt 3540 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca 3600 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat 3660 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg 3720 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg 3780 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca 3840 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg 3900 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa 3960 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa 4020 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca 4080 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt 4140 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt 4200 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg 4260 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac 4320 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac 4380 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg 4440 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg 4500 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt 4560 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 4620 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 4680 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat 4740 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta 4800 gaaataattt tgtttaactt taagaaggag atatacatat gcaccaccac caccaccacg 4860 gctatggccg caaaaaacgc cgccagcgcc gccgcggcta tccgtatgat gtgccggatt 4920 atgccccatg ggatatcatg ccgaaaaaga aacgtaaagt ggggctcgag cccggggaaa 4980 agccctacaa atgccccgaa tgtggtaagt ctttttctag gaacgacacc ttgacagaac 5040 accagcggac ccacaccggg gaaaagccct acaagtgtcc tgagtgcgga aagtctttct 5100 ccactagcgg ttcattagta agacaccaga ggacacacac cggggaaaaa ccgtacaagt 5160 gtcctgagtg cgggaagagt ttctccgatc cgggccactt agtaagacat cagaggacac 5220 ataccgggga gaaaccttat aaatgcccag aatgcgggaa atcgttcagt caaagagcac 5280 atttagaaag acatcaacgg acccacaccg gggaaaagcc ctacaagtgt cctgagtgcg 5340 gaaagtcttt ctccactagc ggttcattag taagacacca gaggacacac accggggaaa 5400 aaccttacaa gtgccctgag tgcggcaaga gcttctctca atcaagttca ttagtaagac 5460 accagaggac tcataccggt ggcggcagcg gcggcagcga attcgggcgc gccgacgcgc 5520 tggacgattt cgatctcgac atgctgggtt ctgatgccct cgatgacttt gacctggata 5580 tgttgggaag cgacgcattg gatgactttg atctggacat gctcggctcc gatgctctgg 5640 acgatttcga tctcgatatg ttaattaacg gatccgagca gaaactcatc tctgaagaag 5700 atctggaaca aaagttgatt tcagaagaag atctggaaca gaagctcatc tctgaggaag 5760 atctgtaagc ggccgcactc gagcaccacc accaccacca ccaccactaa ttgattaata 5820 cctaggctgc taaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 5880 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 5940 ggaactatat ccggat 5956 <210> 89 <211> 5956 <212> DNA <213> Artificial Sequence <220> <223> synthetic construct <400> 89 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc 3180 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc 3240 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3300 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3360 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct 3420 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc 3480 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt 3540 cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca 3600 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat 3660 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg 3720 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg 3780 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca 3840 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg 3900 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa 3960 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa 4020 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca 4080 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt 4140 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt 4200 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg 4260 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac 4320 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac 4380 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg 4440 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg 4500 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt 4560 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag 4620 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct 4680 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat 4740 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta 4800 gaaataattt tgtttaactt taagaaggag atatacatat gcaccaccac caccaccacg 4860 gctatggccg caaaaaacgc cgccagcgcc gccgcggcta tccgtatgat gtgccggatt 4920 atgccccatg ggatatcatg ccgaaaaaga aacgtaaagt ggggctcgag cccggggaga 4980 aaccatacaa atgtcccgaa tgtggcaaga gtttcagcag taaaaagcat ctcgctgagc 5040 atcagagaac tcacaccggg gaaaagccct acaagtgtcc tgagtgcgga aagtctttct 5100 ccactagcgg ttcattagta agacaccaga ggacacacac cggggagaaa ccttataaat 5160 gcccagaatg cgggaaatcg ttcagtcaaa gagcacattt agaaagacat caacggaccc 5220 acaccgggga aaagccatat aaatgccccg agtgcggcaa atcattcagc caaagtagca 5280 acttagtaag acaccagcgc acccataccg gggagaaacc atacaaatgc cccgagtgtg 5340 gaaagtcatt tagtgatcca ggcgcattag taagacatca gcggacacat accggggaaa 5400 agccctacaa gtgtcctgag tgcggaaagt ctttctccac tagcggttca ttagtaagac 5460 accagaggac acacaccggt ggcggcagcg gcggcagcga attcgggcgc gccgacgcgc 5520 tggacgattt cgatctcgac atgctgggtt ctgatgccct cgatgacttt gacctggata 5580 tgttgggaag cgacgcattg gatgactttg atctggacat gctcggctcc gatgctctgg 5640 acgatttcga tctcgatatg ttaattaacg gatccgagca gaaactcatc tctgaagaag 5700 atctggaaca aaagttgatt tcagaagaag atctggaaca gaagctcatc tctgaggaag 5760 atctgtaagc ggccgcactc gagcaccacc accaccacca ccaccactaa ttgattaata 5820 cctaggctgc taaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 5880 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 5940 ggaactatat ccggat 5956

Claims (22)

  1. 활성 단백질 도메인(활성 protein domain) 및 핵국재화서열(nuclear localization sequence)에 융합된 OPA1 유전자 프로모터를 특정적으로 표적하는 폴리닥틸 징크핑거 단백질(polydactyl zinc finger protein)을 포함하는 인공전사인자(artificial transcription factor).
  2. 제1항에 있어서, 단백질 형질도입(transduction) 도메인을 추가적으로 포함하는 것을 특징으로 하는 인공전사인자.
  3. 제1항 또는 제2항에 있어서, 헥사메릭 징크핑거(hexameric zinc finger) 단백질을 포함하는 것을 특징으로 하는 인공전사인자.
  4. 제1항 내지 제3항 중 어느 한 항에 있어서, 상기 활성 단백질 도메인은 서열번호:1의 VP16, 서열번호:2의 VP64, 서열번호:3의 CJ7, 서열번호:4의 p65TA1, 서열번호:5의 SAD, 서열번호:6의 NF-1, 서열번호:7의 AP-2, 서열번호:8의 SP1-A, 서열번호:9의 SP1-B, 서열번호:10의 Oct-1, 서열번호:11의 Oct-2, 서열번호:12의 Oct2-5x, 서열번호:13의 MTF-1, 서열번호:14의 BTEB-2 또는 서열번호:15의 LKLF인 것을 특징으로 하는 인공전사인자.
  5. 제1항 내지 제4항 중 어느 한 항에 있어서, 상기 핵국재화서열(nuclear localization sequences)은 K-K/R-X-K/R 컨센서스(consensus) 서열 또는 서열번호:62의 SV40 NLS를 포함하는 염기(basic) 아미노산의 클러스터(cluster)인 것을 특징으로 하는 인공전사인자.
  6. 제2항 내지 제5항 중 어느 한 항에 있어서, 상기 단백질 형질도입(transduction) 도메인은 서열번호:16의 HIV 유도된 TAT 펩타이드, 서열번호:18의 합성 펩타이드 mT02, 서열번호:19의 합성 펩타이드 mT03, 서열번호:20의 R9 펩타이드, 또는 서열번호:21의 ANTP 도메인인 것을 특징으로 하는 인공전사인자.
  7. 제1항 내지 제6항 중 어느 한 항에 있어서, 서열번호:26 내지 43으로 구성되는 군으로부터 선택되는 단백질 서열의 징크핑거 단백질을 포함하는 것을 특징으로 하는 인공전사인자.
  8. 제1항 내지 제7항 중 어느 한 항에 있어서, 폴리에틸렌 글리콜 잔기(polyethylene glycol residue)를 추가적으로 포함하는 것을 특징으로 하는 인공전사인자.
  9. 제1항 내지 제8항 중 어느 한 항에 따른 인공전사인자를 포함하는 약학적 조성물.
  10. 제1항 내지 제7항 중 어느 한 항에 따른 인공전사인자를 코딩하는 핵산(nucleic acid).
  11. 제10항에 따른 핵산을 포함하는 벡터.
  12. 제11항에 있어서, 상기 벡터는 바이러스 벡터인 것을 특징으로 하는 벡터.
  13. 제11항 또는 제12항에 따른 벡터를 포함하는 숙주 세포(host cell).
  14. 서열번호:83 내지 89의 발현 구성체(expression construct)을 포함하는 제13항에 따른 E. coli 숙주 세포.
  15. 제10항에 따른 핵산을 포함하는 바이러스 담체(viral carrier).
  16. 제15항에 있어서, 아데노 관련 바이러스(adeno-associated viruses), 레트로바이러스(retroviruses), 렌티바이러스(lentiviruses), 아데노바이러스(adenoviruses), 슈도유형 아데노 관련 바이러스(pseudotyped adeno-associated viruses), 슈도유형 레트로바이러스(pseudotyped retroviruses), 슈도유형 렌티바이러스(pseudotyped lentiviruses) 및 슈도유형 아데노바이러스(pseudotyped adenoviruses)로 구성되는 군으로부터 선택되는 것을 특징으로 하는 바이러스 담체.
  17. 제15항 또는 제16항에 따른 바이러스 담체를 포함하는 약학적 조성물.
  18. 제1항 내지 제8항 중 어느 한 항에 있어서, OPA1 유전자 프로모터로부터 발현을 증가시키는 것에 사용을 위한 인공전사인자.
  19. 제10항에 있어서, OPA1 유전자 프로모터로부터 발현을 증가시키는 것에 사용을 위한 핵산.
  20. 제1항 내지 제8항 중 어느 한 항에 있어서, 보통염색체 우성 위축증(autosomal dominant atrophy), 보통염색체 우성 위축증 플러스(autosomal dominant atrophy plus) 및 녹내장(glaucoma)을 치료하는데 사용을 위한 인공전사인자.
  21. 제10항에 있어서, 보통염색체 우성 위축증(autosomal dominant atrophy), 보통염색체 우성 위축증 플러스(autosomal dominant atrophy plus) 및 녹내장(glaucoma)을 치료하는데 사용을 위한 핵산.
  22. 제1항 내지 제8항 중 어느 한 항에 따른 인공전사인자 또는 제10항에 따른 핵산의 치료학적 유효량을 이를 필요로 하는 환자에게 투여하는 것을 포함하는 보통염색체 우성 위축증(autosomal dominant atrophy), 보통염색체 우성 위축증 플러스(autosomal dominant atrophy plus) 또는 녹내장(glaucoma)의 치료 방법.
KR1020157031596A 2013-04-03 2014-04-02 Opa1 단상부족에 의해 원인이 되는 질병 치료를 위한 인공전사인자 KR20160003691A (ko)

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