KR20220104626A - 예비표적화 단백질을 발현하는 균주를 포함하는 생체 내 미생물 추적 플랫폼 - Google Patents

예비표적화 단백질을 발현하는 균주를 포함하는 생체 내 미생물 추적 플랫폼 Download PDF

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KR20220104626A
KR20220104626A KR1020210146377A KR20210146377A KR20220104626A KR 20220104626 A KR20220104626 A KR 20220104626A KR 1020210146377 A KR1020210146377 A KR 1020210146377A KR 20210146377 A KR20210146377 A KR 20210146377A KR 20220104626 A KR20220104626 A KR 20220104626A
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권성영
민정준
홍영진
유성환
김동연
임진희
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Abstract

본 발명은 모노머형 스트렙타비딘을 발현하는 숙주 세포에 관한 것으로서, 본 발명에 따른 숙주 세포는 체내에서 스트렙타비딘을 발현하여, 비오틴화한 진단제로 숙주 세포의 생체 내 분포를 시각화하고 실시간으로 모니터링할 수 있다.

Description

예비표적화 단백질을 발현하는 균주를 포함하는 생체 내 미생물 추적 플랫폼 {In vivo microbial tracking platform comprising a strain expressing pre-targeting protein}
본 발명은 모노머형 스트렙타비딘을 발현하는 균주 및 비오틴화된 화합물을 포함하여, 이에 따라 다양한 경로로 생체 내에 삽입된 박테리아의 분포를 추적하기 위한 생체 내 세포 추적용 조성물에 관한 것이다.
동물에는 위장(GI)관 내, 피부, 다른 상피성 및 조직 틈새, 예컨대 구강, 눈 표면 및 질 내에 공생하는 미생물이 군집화해 있다. 이들 미생물 군집에서 미생물 균주 간 그리고 미생물 및 숙주, 예를 들면 숙주 면역계 간 상호작용이 미생물 집단 구조를 형상화하며, 미생물 분포에 영향을 미치는 자원의 분배 내지 경쟁이 일어난다. 그와 같은 자원은 식품, 위치 및 미생물이 자라기 위한 공간 또는 부착할 수 있는 물리적 구조의 이용 가능성일 수 있다. 예를 들면, 숙주 식사가 위장관의 공생 미생물 군집 및 분포에 관여된다.
반면, 결핵(Mycobacterium tuberculosis), 페스트(Yersinia pestis), 탄저(Bacillus anthracis), 콜레라(Vibrio cholerae), 위궤양(Helicobacter pylori), 폐렴(Streptococcus pneumoniae), 장티푸스(Salmonella typhi), 충치(Streptococcus mutans) 등 인간의 많은 질병이 병원성 미생물과 관련되어 있다.
따라서, 공생 미생물 또는 병원성 미생물과 동물 숙주 사이의 상호 작용은 이전에 알려지지 않은 동물 생리에 영향을 미치는 다양한 메커니즘을 보여줄 수 있을 것으로 기대된다. 상기 메커니즘들은 염증성 장 질환, 비만, 병원체에 대한 내성, 당뇨병 및 신경 퇴행성 질환을 포함한 광범위한 질병 상태와 관련이 있다. 따라서 이러한 상관 관계로 인해 숙주-미생물 상호 작용을 모니터링하기 위한 신뢰할 수 있는 방법이 요구되어져 왔다.
그러나, 종래에는 배설물 및 조직 샘플의 DNA 및 RNA 시퀀싱에서 장내 미생물 군집에 대한 구성을 추측할 수밖에 없었다. 이 접근법은 비침습적 일 수 있으며 많은 박테리아 종에 대한 중요한 정보를 동시에 산출하지만 정확한 정보를 제공하지는 않는다. 반면, 숙주 동물에서 특정 미생물의 시공간적 위치 또는 밀도에 대한 정보를 확인하기 위하여 형광 혼성화(in situ hybridization) 및 수확된 조직에서 미생물의 면역 형광 염색을 실시할 경우, 더 많은 공간 정보를 제공하나, 이러한 방법은 본질적으로 침습적이며 조직 생검이 가능할 경우 또는 작은 동물 연구의 종말점에서만 수행될 수 있어 종단 연구 및 실시간 추적을 방해한다.
또한 상기 방법들은 미생물들의 생체 내 분포를 실시간으로 파악하지는 못한다. 따라서, 상기 공생 미생물 또는 병원성 미생물들이 체내에서 어떻게 분포하는지 비침습적인 방법으로 실시간으로 추적할 수 있다면, 이로운 미생물들에 대한 생산과 병원성 미생물들에 대한 치료 방법을 개발하는 등 다양한 생물학적 응용분야에 적용될 수 있다.
본 발명의 일 목적은 모노머형 스트렙타비딘(monomeric streptavidin; mSA)을 발현하는 숙주 세포 및 선택적으로 비오틴화된 화합물을 포함하는 생체 내 세포 추적용 조성물을 제공하는 것이다.
본 발명의 다른 목적은 모노머형 스트렙타비딘을 발현하는 숙주 세포가 투여된 목적하는 개체에서, 상기 숙주 세포를 촬영 수단으로 확인하는 단계를 포함하는, 생체 내 세포의 분포를 확인하기 위한 정보 제공 방법을 제공하는 것이다.
그러나 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당 업계에서 통상의 지식을 가진 자에게 명확하게 이해될 수 있을 것이다.
이하, 본원에 기재된 다양한 구현예가 도면을 참조로 기재된다. 하기 설명에서, 본 발명의 완전한 이해를 위해서, 다양한 특이적 상세사항, 예컨대, 특이적 형태, 조성물 및 공정 등이 기재되어 있다. 그러나, 특정의 구현예는 이들 특이적 상세 사항 중 하나 이상 없이, 또는 다른 공지된 방법 및 형태와 함께 실행될 수 있다. 다른 예에서, 공지된 공정 및 제조 기술은 본 발명을 불필요하게 모호하게 하지 않게 하기 위해서, 특정의 상세사항으로 기재되지 않는다. "한 가지 구현예" 또는 "구현예"에 대한 본 명세서 전체를 통한 참조는 구현예와 결부되어 기재된 특별한 특징, 형태, 조성 또는 특성이 본 발명의 하나 이상의 구현예에 포함됨을 의미한다. 따라서, 본 명세서 전체에 걸친 다양한 위치에서 표현된 "한 가지 구현예에서" 또는 "구현예"의 상황은 반드시 본 발명의 동일한 구현예를 나타내지는 않는다. 추가로, 특별한 특징, 형태, 조성, 또는 특성은 하나 이상의 구현예에서 어떠한 적합한 방법으로 조합될 수 있다.
본 발명 내 특별한 정의가 없으면 본 명세서에 사용된 모든 과학적 및 기술적인 용어는 본 발명이 속하는 기술분야에서 당업자에 의하여 통상적으로 이해되는 것과 동일한 의미를 가진다.
본 발명의 일 구현 예에 따르면, 모노머형 스트렙타비딘(monomeric streptavidin; mSA)을 코딩하는 유전자가 도입되어 형질 전환된 숙주 세포를 포함하는, 생체 내 세포 추적용 조성물을 제공한다.
본 발명에서 상기 "스트렙타비딘(streptavidin)"은 비오틴(biotin)에 대하여 높은 결합력을 갖는 단백질이며 상기 비오틴과의 특이적인 상호작용을 이용하여 다양한 생물학적 응용분야에 적용되어 왔다. 상기 스트렙타비딘 단백질의 아미노산 서열은 상기 서열번호 1과 같이 나타낼 수 있고, 상기 스트렙타비딘을 코딩하는 유전자는 서열번호 2로 표시되는 것일 수 있으나, 이에 제한되지는 않는다.
본 발명에서 상기 "모노머형 스트렙타비딘(monomeric streptavidin; mSA)"은 상기 스트렙타비딘이 테트라머를 형성하며 바이오틴 컨쥬게이트의 원치 않는 가교 결합을 초래할 수 있도록, 모노머로 존재하는 스트렙타비딘이다.
본 발명에서, 상기 숙주 세포는 말토오즈 결합 단백질(maltose-binding protein; MBP)을 코딩하는 유전자가 더 도입된 것일 수 있다. 상기 "말토오즈 결합 단백질(maltose-binding protein; MBP)"은 대장균의 말토오즈/말토 덱스트린 시스템의 일부로, 말토 덱스트린의 흡수와 효율적인 이화 작용을 담당하는 약 42.5kDa의 단백질이며, 서열번호 3의 아미노산 서열로 나타낼 수 있고, 상기 말토오즈 결합 단백질을 코딩하는 유전자는 서열번호 4로 표시되는 것일 수 있으나, 이에 제한되지는 않는다.
본 발명에서, 상기 숙주 세포는 상기 모노머형 스트렙타비딘을 코딩하는 유전자의 발현을 조절하는 조절 유전자가 더 도입된 것일 수 있다. 상기 "조절" 내지 "발현 조절"은 특정 유전자의 전사 및 번역이 활성화 또는 억제되는 것을 의미할 수 있다.
본 발명에서, 상기 조절 유전자는 구조적으로 DNA-의존 RNA 폴리머라제(DNA-dependent RNA polymerase) 결합 위치, 전사 개시 지점 및 전사 인자 결합 지점, 억제 및 활성 단백질 결합 지점 및 당해 기술 분야의 숙련자에게 직접적으로 혹은 간접적으로 전사량 조절에 작용할 수 있다고 알려진 뉴클레오티드의 다른 어떠한 서열을 포함되는 핵산 분획을 나타낼 수 있으나, 이에 제한되지 않는다.
본 발명에서, 상기 조절 유전자는 상기 모노머형 스트렙타비딘을 코딩하는 유전자의 개시코돈의 5' 상류에 작동가능하도록 연결될 수 있다.
본 발명에서, 상기 조절 유전자는 리보솜 결합 부위(Ribosome binding site; RBS), 5'-비 번역 영역(5'-Untransrated Region; 5'-UTR), 전사 인자 결합 부위(transcription factor binding site) 및 유도성 프로모터(Inducible promoter)로 이루어진 군으로부터 선택되는 적어도 하나인 것일 수 있으나, 이에 제한되는 것은 아니다.
본 발명에서, 상기 "리보솜 결합 부위(Ribosome-binding site; RBS)"는 유전자의 개시코돈의 상류에서 리보솜의 모집을 담당하여 번역을 진행시킨다. 원핵 생물의 리보솜 결합 부위는 5'-AGGAGG-3' 서열을 가지는 샤인-달가노(Shine-Dalgarno; SD) 서열을 포함한다. 상기 샤인-달가노 서열에 16S rRNA의 3' 말단이 상보적으로 결합하여 번역이 시작되는데, 상기 상보적인 서열인 CCUCCU 서열을 항-샤인-달가노(anti-Shine-Dalgarno; ASD) 서열이라고 한다.
본 발명에서, 상기 "5'-비 번역 영역(5'-Untransrated Region; 5'-UTR)"은 5' 지역에서 mRNA의 아미노산으로 번역되는 부분인 코딩 부분(coding region)의 양 쪽에 위치하는 번역이 되지 않는 부분으로, 진화 과정에서 불필요하게 버려지는 부분(junk)로 여겨졌으나, 유전자 발현 조절에 큰 역할을 하는 것으로 알려졌다.
본 발명에서, 상기 "전사 인자 결합 부위(transcription factor binding site)"는 근처의 특정 유전자를 켜거나 끄는 역할을 하는 DNA 부위이다. 상기 전사 인자 결합 부위는 상기 조절 단백질을 암호화하는 유전자의 프로모터, 인핸서(enhancer) 및 사일렌서(silencer)로 이루어진 군으로부터 선택되는 적어도 하나인 것일 수 있으나, 이에 제한되지는 않는다.
본 발명에서 상기 "유도성 프로모터"란, 특정 화학적 또는 물리적 조건 하에서만 특이적으로 하류에 연결된 유전자가 발현될 수 있도록 전사하는 프로모터로서, 예를 들면 IPTG(isopropyl-beta-D-1-thiogalactopyranoside)와 같은 갈락토스의 존재하에서 발현되는 LacZ 유전자의 프로모터, L-아라비노스의 존재 하에서만 발현되는 아라비노스 오페론인 araBAD 프로모터, 테트라사이클린에 의해 발현이 조절되는 tet 프로모터일 수 있고, 바람직하게는 araBAD 프로모터일 수 있다.
본 발명에서, 상기 조절 유전자는 재조합된 상기 벡터가 숙주 세포에 형질 전환되는 경우, 상기 숙주 세포의 주변 세포질로 모노머형 스트렙타비딘이 발현되도록 하는 것이, 상기 숙주 세포의 내부에 머물거나 상기 주변 세포질에 머물지 않고 배출되는 경우보다 발현되는 모노머형 스트렙타비딘의 활용성이 높아 바람직하다.
본 발명에서, 상기 조절 유전자는 서열번호 26 내지 91 중 어느 하나로 표시되는 것일 수 있다.
본 발명에서, 상기 조절 유전자는 총 깁스 자유 에너지 변화량(ΔGtotal)이 0 이하인 것일 수 있다. 상기 "총 깁스 자유에너지 변화량(total Gibbs free energy change; ΔGtotal)"은, 상기 모노머형 스트렙타비딘의 번역 과정에서, 상기 조절 유전자의 mRNA 전사체(transcript)가 리보솜의 30S 서브유닛 복합체 상에 결합하기 전과 후의 깁스 자유 에너지의 차이 값이다. 상기 총 깁스 자유 에너지 변화량(ΔGtotal)이 0 이하인 경우, 상기 모노머형 스트렙타비딘을 코딩하는 유전자의 전사와 번역 능력이 상승할 수 있다. 상기 총 깁스 자유 에너지 변화량(ΔGtotal)은 하기 식 1 및 식 2와 같은 방법으로 계산될 수 있다.
[식 1]
ΔGtotal = (ΔGfinal) - (ΔGinitial)
[식 2]
(ΔGfinal) - (ΔGinitial) = [(ΔGmRNA-rRNA) + (ΔGspacing) + (ΔGstacking) + (ΔGstandby) + (ΔGstart)] - (ΔGmRNA)
상기 식 1 및 식 2에서, 상기 "ΔGfinal"은 리보솜의 30S 서브유닛 복합체가 상기 조절 유전자의 mRNA 전사체 상에 결합한 후의 깁스 자유 에너지 변화량이고; 상기 "ΔGinitial"은 리보솜의 30S 서브유닛 복합체가 상기 조절 유전자의 mRNA 전사체 상에 결합하기 전의 깁스 자유 에너지 변화량이다. 또한, 상기 식 2에서, 상기 "ΔGmRNA-rRNA"는 조절 유전자의 mRNA와 리보솜의 30S 서브유닛의 복합체를 형성하는 반응이 일어날 때의 깁스 자유 에너지 변화량이고, 상기 "ΔGspacing"은 상기 조절 유전자의 mRNA 전사체에서 리보솜의 30S 서브유닛의 복합체를 형성하는 서열과 개시코돈 간의 간격(Spacing)이 최적화 되지 않음으로써 발생하는 대한 깁스 자유 에너지 패널티(penalty)이며, 상기 "ΔGstacking"은 상기 간격(Spacing)의 영역에서 쌓인 뉴클레오티드의 깁스 자유 에너지 변화량이고, 상기 "ΔGstandby"는 상기 조절 유전자의 mRNA 전사체의 대기 부위(Stanby Site)와 리보솜 간 결합 반응이 일어나는 경우 깁스 자유 에너지 패널티, 상기 "ΔGstart"는 mRNA-tRNA 복합체를 형성하는 반응이 일어날 때의 깁스 자유 에너지 변화량, 상기 "ΔGmRNA"는 상기 조절 유전자의 mRNA 전사체가 접힌 복합체 구조물을 형성할 때의 깁스 자유 에너지 변화량이다.
본 발명에서, 상기 각 깁스 자유 에너지 변화량(ΔG)은 NUPACK, ViennaRNA 또는 UNAfold와 같이, 희석된 용액에서 유전자 가닥들의 상호 작용, 농도, 염기쌍의 복잡도, 매듭 구조 등의 변수를 고려하여 계산을 수행하는 소프트웨어에 의해 계산될 수 있으나, 이에 제한되지는 않는다.
본 발명에서, 상기 조절 유전자는 모노머형 스트렙타비딘의 생산량을 극대화할 수 있도록, 번역 개시 속도(Translation Initiation Rate; TIR)가 특정 범위를 갖도록 조절된 것일 수 있다.
본 발명에서 상기 "번역 개시 속도(Translation Initiation Rate; TIR)"는 하기 식 3으로 계산될 수 있으며, 합성 생물학에서 번역 단계는 전체 단백질 생산의 속도를 제한하는 단계이므로 유전자의 발현에 중요한 인자이다.
[식 3]
TIR = exp[k{(ΔGtotal) - (ΔG1total)}]
상기 식 3에서,
TIR의 단위는 au이고;
k는 볼츠만 상수로, 0.4 내지 0.6 mol/kcal일 수 있으며;
ΔGtotal은 상기 식 1에서 정의된 바와 같고;
ΔG1total은 상기 조절 유전자가 포함되지 않은 본 발명의 벡터에서의 깁스 자유 에너지 변화량에 해당하고, 바람직하게는 상기 조절 유전자가 포함되지 않고, 나머지 서열은 동일한 벡터의 자유 에너지 변화량에 해당할 수 있으나, 이에 제한되지는 않는다. 따라서, 상기 조절 유전자가 포함되지 않을 경우에 번역 개시 속도는 1au에 해당한다.
본 발명에서, 상기 조절 유전자는 번역 개시 속도가 50 내지 45000au, 바람직하게는 900 내지 45000au가 되도록 조절된 것이 형질 전환된 균주가 상기 모노머형 스트렙타비딘을 고효율로 생산할 수 있어 바람직하다.
본 발명에서, 상기 조절 유전자의 서열 길이는 15 내지 39bp인 것일 수 있고, 바람직하게는 26 내지 31bp로 이루어 질 수 있으나, 이에 제한되지는 않는다.
본 발명에서, 상기 조절 유전자는 서열번호 5로 표시되는 유전자 서열인 "AGG"를 포함하는 것일 수 있고, 상기 조절 유전자는 서열번호 6으로 표시되는 유전자 서열인 "TAGG"를 포함하는 것일 수 있으며, 상기 조절 유전자는 서열번호 7로 표시되는 유전자 서열인 "ATAGG"을 포함하는 것일 수 있으나, 이에 제한되지는 않는다.
본 발명에서, 상기 조절 유전자는 서열번호 5 내지 7로 표시되는 유전자 서열의 3' 말단에서 개시코돈 까지의 간격(Spacing)이 6 내지 13bp, 바람직하게는 6 내지 10bp인 것일 수 있다. 상기 간격이 6 내지 13bp인 경우, 상기 mRNA 전사체에서 rRNA 복합체를 형성하는 서열과 개시코돈 간의 최적화되지 않은 간격(Spacing)에 대한 깁스 자유 에너지 패널티(ΔGspacing)가 최소화되어, 그 결과 모노머형 스트렙타비딘의 발현 수준을 높일 수 있다.
본 발명의 바람직한 일 예시에서, 상기 조절 유전자는 상기 식 1의 총 깁스 자유 에너지 변화량(ΔGtotal)이 0 이하이고, 번역 개시 속도(TIR)가 900 내지 9000au로 조절된 것으로, 서열 길이는 26 내지 31bp로 이루어진 것이며, 서열번호 5 내지 7 중 어느 하나로 표시되는 유전자 서열을 포함하고, 상기 서열번호 5 내지 7 중 어느 하나로 표시되는 유전자의 3' 말단으로부터 상기 모노머형 스트렙타비딘을 코딩하는 유전자의 개시코돈까지 간격(Spacing)이 6 내지 10bp일 수 있다.
본 발명의 바람직한 다른 일 예시에서, 상기 조절 유전자는 서열번호 32 또는 36으로 표시될 수 있다.
본 발명의 형질 전환은 벡터에 의해 이루어질 수 있다. 상기 "벡터(vector)"내지 유전자 컨스트럭트는 외래 유전자를 세포로 전달하여 발현할 수 있는 수단으로, 본 발명의 상기 벡터는 플라스미드(plasmids), 코스미드(cosmids), 인공 염색체(artificial chromosomes), 리포솜(Liposomes)과 같은 비바이러스성 벡터이거나, 레트로바이러스(retrovirus), 아데노바이러스(Adenovirus), 아데노-관련 바이러스(Adenovirus-associated virus; AAV), 파지(phage)와 같은 바이러스성 벡터일 수 있다.
본 발명에서 상기 "플라스미드(plasmids)"는 염색체와 분리되어 있고 자신의 복제기점을 소유하여 독립적으로 증식 가능한 에피솜 DNA 분자이다. 상기 플라스미드는 제한효소에 의해 재조합된 후 숙주 세포에 전달되어 벡터의 기능을 할 수 있다.
본 발명에서 상기 "코스미드(cosmids)"는 파이 파지의 점착성 말단(cohesive ends)인 cos 부위를 이용한 플라스미드로, 삽입할 수 있는 유전자의 크기가 커서 유전자 라이브러리를 만드는데 주로 사용된다.
본 발명에서 상기 "인공 염색체(artificial chromosomes)"는 벡터로 사용하기 위하여 인위적으로 구조를 변화시킨 염색체로, 박테리아 인공염색체, 효모 인공염색체, 인간 인공염색체 등이 있다.
본 발명에서 상기 "리포솜(Liposomes)"은 인위적으로 만든 1개 이상의 지질 2중층으로 되어 있는 소낭 구조물로, 세포막의 형태와 유사하고 다양한 물질을 혼입하는 능력 때문에 핵산뿐만 아니라 펩티드, 항체, 앱타머 등을 전달하는 약물 운반체 시스템이다. 상기 리포솜의 효능은 막과 성분의 특성에 따른 표적 전달 및 침투 능력에 달려 있다.
본 발명에서 상기 "레트로바이러스(retrovirus)"는 역전사(reverse transcription)를 통한 DNA 중간체를 필수로 하는 단일가닥의 양성 센스(positive-sense) RNA를 게놈으로 가지는 바이러스로, 레트로바이러스 벡터는 숙주 세포의 염색체에 삽입되어 세포분열을 하여도 바이러스 벡터가 안정적으로 유지되므로 유전자 치료에 많이 사용된다.
본 발명에서, 상기 "렌티바이러스(Lentivirus)"는 레트로바이러스의 일종으로, 숙주에 내인성인 바이러스(endogenous retrovirus; ERV)이다. 비리온 입자는 약간 다형성의 직경 80 내지 100nm 구형이고, 뉴클레오캡시드(코어)는 등척성이며, 뉴클레오티드는 동심 막대 형상 또는 원추 형상이다.
본 발명에서 상기 "아데노바이러스(Adenovirus)"는 약 36kb DNA를 가지는 바이러스로, 50가지 이상의 유전자를 가지고 있어 몇 가지의 바이러스 유전자를 발현하려는 유전자로 치환하여 벡터를 생성할 수 있다.
본 발명에서 상기 "아데노바이러스-관련 바이러스(Adenovirus-associated virus; AAV)"는 DNA 유전체가 매우 작고, 아데노바이러스가 필요한 인공위성바이러스(Satellite virus)로, 벡터로 사용할 경우 사람 염색체의 특정한 부위에 삽입되어 잠복감염(latent infection)을 일으킨다.
본 발명에서, 상기 재조합 벡터는 구성적 발현 벡터(Constitutive Expression Vector)나 유도성 발현 벡터(Inducible Expression Vector) 일 수 있고, 그 예시로 pKD13, pCP20, pMA1, pUC19, pJL, pBAD, pET, pGEX, pMAL, pALTER, pCal, pcDNA, pDUAL, pTrc, pQE, pTet, pProEX HT, pPROLar.A, pPROTet.E, pRSET, pSE280, pSE380, pSE420, pThioHis, pTriEx, pTrxFus, Split GFP Fold ′n' Glow, pACYCDuet-1, pCDF-1b, pCDFDuet-1, pCOLADuet-1, pLysS, pRSF-1b, pRSFDuet-1, pT7-FLAG, T7Select, pCMV, pBluescript, pBac, pAc, pFastBacHT, pFastBac, pAO815, pPIC, pESC, pCas9, pwtCas9-bacteria, pgRNA-bacteria 및 pGRG 플라스미드 또는 에서 선택된 1 이상의 플라스미드에서 유래된 것일 수 있으나, 이에 제한되지는 않는다.
본 발명의 일 예시에서, 상기 pKD13은 약 3.4kbp로 이루어져 있고, 베타-락타마제(beta-lactamase), Tn5 네오마이신 포스포트랜스퍼라제(Tn5 neomycin phosphotransferase), 람다 종결자(lambda terminator) 및 R6K 감마 복제 원점(R6K gamma replication origin) 유전자를 포함할 수 있다.
본 발명의 일 예시에서, 상기 pCP20 플라스미드는 약 9.4kbp로 이루어져 있고, EcoRI, cat, Pstl, HindⅢ, Ci857, flp, bamHi, 베타-락타마제(beta-lactamase), mobA, mob2 및 repA101ts 유전자 부위를 포함할 수 있다.
본 발명의 일 예시에서, 상기 pMA1 플라스미드는 Microcystis aeruginosa f. aeruginosa Kutzing에서 유래하고, 약 2.3kbp로 이루어져 있으며, HincⅡ 유전자 부위를 포함하는 것일 수 있다.
본 발명의 일 예시에서, 상기 pJL 플라스미드는 빈 백본을 가지고 RNA 바이러스 기반인 것일 수 있다.
본 발명의 일 예시에서, 상기 pBAD, pCMV 및 pCMV 플라스미드는 포유류 숙주 세포에서 발현되고 CMV, 프로모터를 이용하며, 암피실린내성을 가지는 것일 수 있다.
본 발명의 일 예시에서, 상기 pET, pBluescript, pCal 및 pcDNA 플라스미드는 박테리아 숙주 세포에서 발현되고 T7 또는 Lac 프로모터를 이용하며, 암피실린 내성을 가지는 것일 수 있다.
본 발명의 일 예시에서, 상기 pMAL 및 pGEX 플라스미드는 박테리아 숙주 세포에서 발현되고 Tac 프로모터를 이용하며, 암피실린 내성을 가지는 것일 수 있다.
본 발명의 일 예시에서, 상기 pALTER 플라스미드는 박테리아 숙주 세포에서 발현되고 T7 프로모터를 이용하며, 테트라사이클린 내성을 가지는 것일 수 있다.
본 발명의 일 예시에서, 상기 pDUAL 플라스미드는 박테리아 숙주 세포에서 발현되고 T7 또는 Lac 프로모터를 이용하며, 카나마이신 내성을 가지는 것일 수 있다.
본 발명의 일 예시에서, 상기 pTrc 플라스미드는 박테리아 숙주 세포에서 발현되고 trc 프로모터를 이용하며, 암피실린 내성을 가지는 것일 수 있다.
본 발명의 일 예시에서, 상기 pUC19 플라스미드는 박테리아 숙주 세포에서 발현되고, 약 2.6kbp의 원형 이중 가닥 DNA로 이루어져 있으며, pUC18과 MCS 영역이 반대인 벡터이다. 상기 pU19 벡터는 형질 전환을 위하여 가장 널리 사용되는데, 상기 pU19에 의해 외래 DNA가 도입된 숙주 세포는 대조군에 비하여 성장 배지 속 콜로니의 색상이 달라지므로, 구별이 가능하다.
본 발명의 일 예시에서, 상기 pQE 플라스미드는 T5-lac 프로모터를 이용하고, 암피실린 내성을 가지는 것일 수 있다.
본 발명의 일 예시에서, 상기 pTet 플라스미드는 tet 오페론으로부터의 조절 서열의 제어 하에 CMV 프로모터를 포함하고 있어, 트랜스 활성화제인 pTet-tTAk와 공동 형질 감염될 때 독시사이클린이 없는 경우에만 단백질을 발하는 것일 수 있다.
본 발명의 일 예시에서, 상기 pCas9, pwtCas9-bacteria 및 pgRNA-bacteria 플라스미드는 CRISPR 기술을 사용하여, Cas9 뉴클레아제 gRNA을 발현시키기 위한 것일 수 있다.
본 발명에서, 상기 숙주 세포를 형질 전환시키는 방법은 당업계의 통상적인 도입 방법에 따라 수행될 수 있고, 구체적인 방법을 특별히 제한하지는 않으나, 예를 들면, 박테리아의 형질 전환 방법, CaCl2 침전법, CaCl2 방법에 환원물질인 DMSO(Dimethyl sulfoxide)를 사용함으로써 효율을 높인 Hanahan 방법, 전기천공법(Electroporation), 인산칼슘 침전법, 원형질 융합법, 실리콘 카바이드 섬유를 이용한 교반법, 아그로 박테리아 매개된 형질 전환법, PEG를 이용한 형질 전환법, 덱스트란 설페이트, 리포펙타민 및 건조/억제 매개된 형질 전환 방법 등을 사용할 수 있다.
본 발명에서, 상기 숙주 세포를 생체 내에 투여하는 경우 생체 내에서 상기 숙주 세포의 분포를 확인할 수 있는 효과가 있다.
본 발명에서, 상기 숙주 세포는 박테리아, 효모, 균류 세포, 식물 세포, 곤충 세포 및 동물 세포로 이루어진 군으로부터 선택되는 어느 하나 이상의 세포인 것일 수 있다.
본 발명에서 상기 박테리아는, 락토코쿠스(Lactococcus), 류코노스톡(Leuconostoc), 페디오코커스(Pediococcus), 엔트로코커스(Enterococcus), 스트렙토코커스(Streptococcus), 베일로넬라(Veilonella), 에스체리시아(Escherichia), 유박테리움(Eubacterium), 슈도모나스(Pseudomonas), 살모넬라(Salmonella), 시겔라(Shigella), 헬리코박터(Helicobacter), 캄필로박터(Campylobacter), 어시니아(Yersinia), 리스테리아(Listeria), 스트렙토미세스(Streptomyces), 펩토코커스(Peptococcus), 펩토스트렙토코커스(Peptostreptococcus), 프로투스(Proteus), 루미노코커스(Ruminococcus), 엔테로박터(Enterobacter), 시트로박터(Citrobacter), 세라티아(Serratia), 해모필러브(Haemophilus), 스타필로코커스(Staphylococcus), 마이코박테리움(Mycobacterium), 클로스트리디움(Clostridium), 바실러스(Bacillus), 마이크로코커스(Micrococcus), 비브리오(Vibrio), 박테로이데스(Bacteroides), 멜리소코코스(Melissococcus), 코쿠리아(Kocuria), 아에로코코스(Aerococcus), 오에노코코스(Oenococcus), 락토바실러스(Lactobacillus), 스포로락토바실러스(Sporolactobacilus), 아커만시아(Akkermansia), 비피도박테리움(Bifidobacterium), 부티리치코쿠스(Butyricicoccus), 부티리시모나스(Butyricimonas), 부티비브리오(Butyrivibrio), 슈도부티비브리오(Pseudobutyrivibrio), 웨이셀라(Weissella), 푸소박테리움(Fusobacterium), 카르노박테리움(Carnobacterium), 프로피오니박테리움(Propionibacterium), 메가스파에라(Megasphaera), 알리스팁스(alistipes), 알로바쿨룸(allobaculum), 바네시엘라(barnesiella), 블라우티아(blautia), 도레아(dorea), 헤스펠리아(hespellia), 홀데마니아(holdemania), 라우소니아(lawsonia), 오실리박터(oscillibacter), 파라박테로이드(parabacteroides), 파스콜락토박테리움(phascolarctobacterium), 프리보텔라(prevotella), 세디멘티박터(sedimentibacter), 엑시구오박데리움(Exiguobacterium), 아시네토박터(Acinetobacter), 카프노시토파가(Capnocytophaga), 네이세리아(Neisseria), 스핑고모나스(Sphingomonas), 아그레가티박터(Aggregatibacter), 렙토트리키아(Leptotrichia), 그라눌리카텔라(Granulicatella), 크리세오박테리움(Chryseobacterium), 포르피로모나스(Porphyromonas), 브라키박테리움(Brachybacterium), 엔하이드로박터(Enhydrobacter), 파라코커스(Paracoccus), 코리네박테리움(Corynebacterium), 로티아(Rothia), 엑티노마이세스(Actinomyces), 디알리스터(Dialister), 페칼리박테리움(Faecalibacterium), 할로모나스(Halomonas), 수테렐라(Sutterella), 베일로넬라(Veillonella), 로도코커스(Rhodococcus), 아토포비움(Atopobium), 크로모할로박터(Chromohalobacter), 쿠프리아비두스(Cupriavidus), 메타노브레비박터(Methanobrevibacter), 오도리박터(Odoribacter), 피라미도박터(Pyramidobacter), 빌로필라(Bilophila), 데설포비브리오(Desulfovibrio), 아시다미노코커스(Acidaminococcus), 아크로모박터(Achromobacter), 아그로박테리움(Agrobacterium), 로시텔레스(Roseateles), 코프로코커스(Coprococcus), 터리시박터(Turicibacter), 로즈뷰리아(Roseburia), 라크노스피라(Lachnospira), 오스실로스피라(Oscillospira), SMB53, 카테니박테리움(Catenibacterium), 파라프레보텔라(Paraprevotella), 아들러크레우치아(Adlercreutzia), 슬라키아(Slackia) 및 데르모아나데로박테리움(Thermoanaerobacterium)에서 선택된 어느 하나 이상일 수 있으나 이에 제한되지는 않는다.
본 발명에서 상기 효모는, 사카로마이세스(Saccharomyces), 데바로마이세스(Debaromyces), 칸디다(Candida), 클루이베로마이세스(Kluyveromyces), 피치아(Pichia), 토룰라스포라(Torulaspora) 및 파피아(Phaffia)에서 선택된 어느 하나 이상일 수 있으나 이에 제한되지는 않는다.
본 발명에서, 상기 균류는 아스페르길루스(Aspergillus), 리조푸스(Rhizopus), 뮤코르(Mucor), 페니실리움(Penicillium) 및 바시디오마이코타(Basidiomycota)에서 선택된 어느 하나 이상일 수 있으나 이에 제한되지는 않는다.
본 발명에서, 상기 곤충 세포는 드로조필라 및 스포도프테라 Sf9 세포에서 선택된 어느 하나 이상일 수 있으나 이에 제한되지는 않는다.
본 발명에서, 상기 동물 세포는 CHO(중국 햄스터 난소 세포, Chinese hamster ovary cells), SP2/0(생쥐 골수종), 인간 림프아구(Human lymphoblastoid), COS, NSO(생쥐 골수종), 293T 세포, 보우 멜라노마 세포, HT-1080 세포, BHK 세포(베이비 햄스터 신장세포, Baby Hamster Kidney cells), HEK 세포(인간 배아신장 세포, Human Embryonic Kidney cells) 및 PERC.6 세포(인간 망막 세포)에서 선택된 어느 하나 이상일 수 있으나 이에 제한되지는 않는다.
본 발명의 일 예시에서, 상기 박테리아는 aroA, aroC, aroD, aroE, Rpur, htrA, ompR, ompF, ompC, galE, cya, crp, cyp, phoP, phoQ, rfaY, dksA, hupA, sipC, clpB, clpP, clpX, pab, nadA, pncB, pmi, rpsL, hemA, rfc, poxA, galU, cdt, pur, ssa, guaA, guaB, fliD, flgK, flgL, relA, spoA 및 spoT로 이루어진 군으로부터 선택되는 적어도 하나를 코딩하는 유전자가 변형되어 발현된 것일 수 있다.
본 발명의 다른 일 예시에서, 상기 박테리아는 구아노신 다인산 합성능이 결여되어 약독화된 것일 수 있다. 상기 구아노신 다인산은 구아노신-5-이인산-3-이인산(ppGpp)일 수 있고, 상기 숙주 세포는, 구아노신-5-이인산-3-이인산(ppGpp)을 가수분해하는 relA 또는 구아노신-5-이인산-3-이인산(ppGpp)을 합성하는 spoT 코딩하는 유전자가 변형되어 상기 구아노신-5-이인산-3-이인산(ppGpp)의 합성능이 결여된 것일 수 있으나, 이에 제한되는 것은 아니다.
본 발명에서, 상기 박테리아에 유전자에 변형을 가하는 방법은 당업계에 공지된 다양한 유전자의 결실 또는 파괴하는 방법에 의해 수행될 수 있고, 예를 들면, 상기 결실 및 파괴 방법은 상동성 재조합, 화학적 변이유발, 조사 변이유발 또는 트랜스포존 변이유발 등과 같은 방법에 의해 수행될 수 있으나, 이에 제한되는 것은 아니다.
본 발명의 조성물은 비오틴화된 화합물을 더 포함할 수 있고, 이때 상기 숙주 세포에서 발현되는 모노머형 스트렙타비딘은 비오틴-결합성 단백질로서, 비오틴과 상호 작용할 수 있는 결합 부위를 포함할 수 있다.
본 발명에서, 상기 비오틴화된 화합물은 비오틴, 바람직하게는 D-비오틴과 반응하도록 적어도 하나의 아민기를 포함하는 화합물에, D-비오틴의 N-히드록시숙신이미딜 에스테르(NHS-비오틴) 와 같은 일반적인 비오틴화된 시약을 이용하여 비오틴기를 갖도록 개질된 것일 수 있다.
본 발명에서, 상기 비오틴화된 화합물은 상기 숙주 세포에서 발현되는 모노머형 스트렙타비딘을 예비 표적화 할 수 있고, 이에 따라 상기 숙주 세포, 또는 상기 숙주 세포로부터 발현되는 모노머형 스트렙타비딘과, 비오틴화된 화합물을 생체 내에 투여하는 경우, 생체 체내의 숙주 세포가 위치한 곳에서 높은 효율로 화합물-비오틴-스트렙타비딘 복합체가 형성될 수 있다.
본 발명에서, 상기 비오틴화된 화합물은 비오틴화된 조영제일 수 있다. 여기서, 상기 조영제는 방사성 핵종, 형광 표지, 효소 표지, 화학 발광 마커, 금 제제 및 자성 제제로 이루어진 군으로부터 선택되는 적어도 하나인 것일 수 있으나, 알파선, 감마선, 양전자(Positron), X-선, 자외선, 가시광선, 적외선, 초음파(ultrasonic wave) 및 자기 공명(magnetic resonance)에서 선택된 하나 이상의 신호를 발생하거나 증폭시키는 것이라면 어떤 것이든 제한되지는 않는다.
본 발명에서, 상기 방사성 동위원소 내지 방사성 핵종은 C-11, F-18, Cu-64, N-13, Ga-68, Sc-44, Zr-89, Y-90, Tc-99m, In-111, I-123, I-124, I-125, I-131 및 Lu-177으로 이루어진 군에서 선택된 어느 하나 이상인 것일 수 있으나, 이에 제한되지는 않는다.
본 발명에서, 상기 방사성 동위원소 내지 방사성 핵종을 검출할 수 있는 수단은, 양전자방출단층촬영(PET) 또는 단일광자방출전산화 단층촬영(SPECT) 일 수 있으나, 이에 제한되지는 않는다. 상기 양전자방출단층촬영(PET)은 양전자를 방출하는 방사성 동위원소를 결합한 약물(방사성의약품)을 체내에 주입하면체내에서 방출된 양전자가 인접한 전자와 결합하여 소멸(annihilation)하면서 2개의 광자를 발생하고 이 2개의 광자를 검출하여 영상을 구성하는 방법이다. 상기 단일 광자방출전산화단층촬영(SPECT)은 단일 광자(감마선)를 발생하는 방사성동위원소를 결합한 약물을 체내에 주입하면 체내에서 방출되는 감마선을 검출하여 영상을 구성하는 방법이다.
본 발명에서, 상기 형광 표지는 플루오레세인, 로다민, 로돌, 로사민 및 이들의 유도체를 비제한적으로 포함하는 피페라진, 크산텐, 쿠마린, 아크리딘, 푸란, 인돌, 퀴놀린, 시아닌, 벤조푸란, 퀴나졸리논, 벤자졸, 붕소-디피로메텐 및 이들의 유도체로 이루어진 군으로부터 선택된 어느 하나 이상일 수 있으나, 이에 제한되지는 않는다.
본 발명에서, 상기 효소 표지는 양고추냉이 퍼옥시다제(Horseradish peroxidase; HRP), 루시퍼라제 및 알칼리성 포스파타제로 이루어진 군에서 선택된 어느 하나 이상인 것일 수 있으나, 이에 제한되지는 않는다.
본 발명에서 상기 "생체 내"란, 상기 숙주 세포가 감염 내지 주입될 수 있는 개체의 생체 내로서, 상기 개체는 포유동물 및 비-포유동물을 모두 포함할 수 있다. 여기서, 상기 포유동물의 예로는 인간, 비-인간 영장류, 예컨대 침팬지, 다른 유인원 또는 원숭이 종; 축산 동물, 예컨대 소, 말, 양, 염소, 돼지; 사육 동물, 예컨대 토끼, 개 또는 고양이; 실험 동물, 예를 들어 설치류, 예컨대 래트, 마우스 또는 기니아 피그 등을 포함할 수 있으나, 이에 제한되는 것은 아니다. 또한, 본 발명에서 상기 비-포유동물의 예로는 조류 또는 어류 등을 포함할 수 있으나, 이에 제한되는 것은 아니다.
본 발명에서 상기 "투여"란, 임의의 적절한 방법으로 개체에게 본 발명의 유효성분을 도입하는 과정을 의미하는 것으로서, 상기와 같이 투여되는 화합물의 제제는 특별히 제한하지 않으며, 고체 형태의 제제, 액체 형태의 제제 또는 흡인용 에어로졸 제제로 투여될 수 있으며, 사용하기 바로 전에 경구 또는 비경구 투여용 액체 형태 제제로 전환되도록 의도되는 고체 형태 제제로 투여될 수 있고, 예를 들면, 산제, 과립제, 캡슐, 정제, 수성 현탁액 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 투여될 수 있으나, 이에 제한되는 것은 아니다.
또한, 본 발명에서 상기 투여 시 본 발명의 숙주 세포 내지 화합물과 함께 약학적으로 허용 가능한 담체를 추가로 투여할 수 있다. 여기서, 상기 약학적으로 허용되는 담체는 경구 투여 시에는 결합제, 활탁제, 붕해제, 부형제, 가용화제, 분산제, 안정화제, 현탁화제, 색소, 향료 등을 사용할 수 있으며, 주사제의 경우에는 완충제, 보존제, 무통화제, 가용화제, 등장제, 안정화제 등을 혼합하여 사용할 수 있으며, 국소투여용의 경우에는 기제, 부형제, 윤활제, 보존제 등을 사용할 수 있다. 본 발명의 화합물의 제형은 상술한 바와 같은 약학적으로 허용되는 담체와 혼합하여 다양하게 제조될 수 있다. 예를 들어, 경구 투여시에는 정제, 트로키, 캡슐, 엘릭서(elixir), 서스펜션, 시럽, 웨이퍼 등의 형태로 제조할 수 있으며, 주사제의 경우에는 단위 투약 앰플 또는 다수회 투약 형태로 제조할 수 있다. 기타, 용액, 현탁액, 정제, 캡슐, 서방형 제제 등으로 제형할 수 있다.
한편, 제제화에 적합한 담체, 부형제 및 희석제의 예로는, 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말디톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로즈, 폴리비닐피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 또는 광물유 등이 사용될 수 있다. 또한, 충진제, 항응집제, 윤활제, 습윤제, 향료, 유화제, 방부제 등을 추가로 포함할 수 있다.
본 발명에 따른 화합물의 투여 경로는 이들로 한정되는 것은 아니지만 구강, 정맥 내, 근육 내, 동맥 내, 골수 내, 경막 내, 심장 내, 경피, 피하, 복강 내, 비강 내, 장관, 국소, 설하 또는 직장이 포함된다. 경구 또는 비경구 투하가 바람직하다.
본 발명에서, "비경구"는 피하, 피내, 정맥내, 근육내, 관절내, 활액낭내, 흉골내, 경막내, 병소내 및 두개골내 주사 또는 주입기술을 포함한다. 본 발명의 조성물은 또한 직장 투여를 위한 좌제의 형태로 투여될 수 있다.
본 발명에서, "유효량"은 바람직한 생물학적 결과를 제공하기 위한 작용제의 충분한 양을 지칭한다. 상기 결과는 생체 내 숙주 세포의 분포를 실시간으로 확인하거나 임의의 다른 바람직한 변화를 이끌어 내는 것일 수 있다. 예를 들어, 숙수 세포를 추적하기 위한 "유효량"은 생체 내에서 숙주 세포를 추적하기 위하여 유의한 양의 신호를 발생하는데 요구되는, 본 발명에 개시된 화합물의 양이다. 임의의 개별적인 경우에서 적절한 "효과적인" 양은 일상적인 실험을 사용하여 당업자에 의해 결정될 수 있다. 본 발명의 경우에, 활성 물질은, 생체 내 세포 추적용 조성물이다.
본 발명에 숙주 세포 내지 화합물의 유효량은 사용된 숙주 세포의 종류, 사용된 특정 화합물의 활성, 개체의 연령, 체중, 일반적인 건강, 성별, 정식, 투여 시간, 투여 경로, 배출율 및 배합을 포함한 여러 요인에 따라 다양하게 변할 수 있지만 당업자에 의해 적절하게 선택될 수 있고, 1일 0.0001 내지 100mg/kg 또는 0.001 내지 100mg/kg으로 투여할 수 있다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. 본 발명에 따른 숙주 세포 내지 화합물은 환제, 당의정, 캡슐, 액제, 겔, 시럽, 슬러리, 현탁제로 제형될 수 있다.
본 발명의 다른 구현 예에 따르면, 모노머형 스트렙타비딘을 발현하는 숙주 세포가 투여된 목적하는 개체에서, 상기 숙주 세포를 촬영 수단으로 확인하는 단계를 포함하는, 생체 내 세포의 분포를 확인하기 위한 정보 제공 방법을 제공한다.
본 발명에서, 상기 목적하는 개체는 비오틴화된 화합물 또한 투여된 것일 수 있다. 상기 비오틴화된 화합물은 모노머형 스트렙타비딘이 발현된 숙주 세포에 결합하여, 상기 숙주 세포의 생체 내 분포를 확인할 수 있는 효과를 가진다.
본 발명에서 상기 촬영 수단은, 알파선, 감마선, 양전자(Positron), X-선, 자외선, 가시광선, 적외선, 초음파(ultrasonic wave) 및 자기 공명(magnetic resonance)에서 선택된 하나 이상의 신호를 이용한 것일 수 있으나, 상기 신호는 목적하는 개체에 비침습적이거나 낮은 침습성을 가지고, 상기 비오틴화된 화합물에 의해 발생되거나 증폭되는 것이라면 제한없이 포함될 수 있다.
본 발명에서, 상기 확인하는 단계 시 상기 비오틴화된 화합물에 의해 발생되거나 증폭되는 상기 신호의 여부, 범위 또는 크기를 상기 촬영 수단에 의해 측정할 수 있다.
본 발명에서, 상기 목적하는 개체는 아라비노스 또한 투여된 것일 수 있다. 상기 아라비노스는 상기 숙주 세포의 모노머형 스트렙타비딘을 지속적으로 발현할 수 있게 하여, 상기 숙주 세포의 생체 내 분포를 확인할 수 있는 효과를 가진다.
또한, 본 발명에서, 상기 확인하는 단계는 1회 또는 복수 회에 걸쳐 수행될 수 있다. 본 발명에서는 특정 시점을 기준으로 복수 회에 걸쳐 확인하는 단계를 수행할 수 있고, 혹은 소정의 기간에 걸쳐 복수 회로 확인하는 단계를 수행하여, 시간에 따른 상기 숙주 세포의 생체 내 분포를 확인할 수 있다.
본 발명의 정보 제공 방법에서, 상기 숙주 세포, 개체 및 비오틴화된 화합물에 관한 기재는 앞서 기재된 바와 중복되어 명세서의 과도한 복잡을 피하기 위하여 이하 그 자세한 기재를 생략한다.
본 발명에 따라 모노머형 스트렙타비딘을 발현하는 숙주 세포를 개체에 투여하는 경우, 상기 숙주 세포로부터 발현되는 모노머형 스트렙타비딘은 생체 내에서 그 기능성을 유지할 수 있다. 이를 통해 비오틴화된 영상제제를 투여함으로써 생체 내 상기 숙주 세포의 존재 및 분포를 확인할 수 있다.
도 1은 실험예 1에서 mSA 유전자를 단독으로 형질 도입시킨 플라스미드의 발현 결과를 나타낸 것이다.
도 2는 실험예 2에서 MBP-mSA 유전자 발현 결과를 나타낸 것이다.
도 3은 실험예 2에서 MBP-mSA 유전자 발현 및 활성 확인을 위한 웨스턴 블롯 결과를 나타낸 것이다.
도 4는 실험예 2에서 재조합 균주의 비오틴 결합을 분석한 결과를 그래프로 나타낸 것이다.
도 5는 실험예 2에서 재조합 균주의 비오틴 결합을 공초점 현미경을 통해 이미지로 나타낸 것이다.
도 6은 실험예 2에서 재조합 균주의 비오틴 결합을 공초점 현미경을 통해 이미지로 나타낸 것이다.
도 7은 실험예 3에서 MBP-mSA 유전자 발현 결과를 나타낸 것이다.
도 8은 실험예 3에서 MBP-mSA 유전자 발현 및 활성 확인을 위한 웨스턴 블롯 결과를 나타낸 것이다.
도 9는 실험예 4에서 MBP-mSA 유전자 발현 확인을 위한 웨스턴 블롯 결과를 나타낸 것이다.
도 10은 실험예 4에서 MBP-mSA 유전자 발현 비교를 위한 웨스턴 블롯 결과를 나타낸 것이다.
도 11은 실험예 4에서 재조합 균주의 비오틴 결합을 분석한 결과를 그래프로 나타낸 것이다.
도 12는 실험예 4에서 재조합 균주의 비오틴 결합을 공초점 현미경을 통해 이미지로 나타낸 것이다.
도 13은 실험예 4에서 재조합 균주의 비오틴 결합을 공초점 현미경을 통해 이미지로 나타낸 것이다.
도 14는 실험예 5에서 재조합 균주의 비오틴 결합 특이성을 분석한 결과를 그래프로 나타낸 것이다.
도 15는 실험예 5에서 재조합 균주의 비오틴 결합 특이성을 분석한 결과를 그래프로 나타낸 것이다.
도 16은 실험예 5에서 마우스의 근육으로 주입된 재조합 균주의 비오틴 결합을 이미지로 나타낸 것이다.
도 17은 실험예 5에서 마우스의 근육으로 주입된 재조합 균주의 개체수를 나타낸 것이다.
도 18은 실험예 5에서 마우스 복강으로 주입된 재조합 균주의 비오틴 결합을 이미지로 나타낸 것이다.
도 19는 실험예 5에서 마우스 복강으로 주입된 재조합 균주의 장관에서 개체수를 나타낸 것이다.
도 20은 실험예 5에서 마우스 정맥으로 주입된 재조합 균주의 비오틴 결합을 이미지로 나타낸 것이다.
도 21은 실험예 5에서 마우스 정맥으로 주입된 재조합 균주의 간에서 개체수를 나타낸 것이다.
도 22는 실험예 5에서 마우스에 경구투여된 재조합 균주의 비오틴 결합을 이미지로 나타낸 것이다.
도 23은 실험예 5에서 마우스에 경구투여된 재조합 균주의 장관에서 개체수를 나타낸 것이다.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.
[실시예 1] mSA 발현 플라스미드 제작
[1-1] mSA 유전자 삽입 플라스미드 제작
재조합 균주에서 mSA 발현을 위한 플라스미드 제작을 위해 서열번호 2로 표시되는 모노머형 스트렙타비딘(mSA) 유전자를 합성(마크로젠, 한국)하여 증폭한 뒤, 제한효소 EcoRⅠ 및 SalⅠ을 넣어 절단하고 이를 정제하여 유전자 증폭 산물을 얻은 뒤에 이를 동일한 제한효소로 절단한 pBAD24 플라스미드에 클로닝 하여 pBAD-mSA(B-mSA) 플라스미드를 제작하였다.
추가로 mSA 유전자의 발현을 증가시키기 위해 프로모터 하류에 하기 표 1의 리보솜 결합 부위(Ribosome Binding Site; RBS)인 BBa_B0032, BBa_B0030, BBa_B0034를 삽입하여 pBAD_RBS 0.3-mSA(B_R0.3-mSA), pBAD_RBS 0.6-mSA(B_R0.6-mSA), pBAD_RBS 1.0-mSA(B_R1.0-mSA) 플라스미드를 제작하였다.
[1-2] MBP-mSA- 발현 플라스미드 제작
그 다음, 동물실험에 이용할 수 있도록 pBAD24 플라스미드에, 서열번호 4로 표시되는 말토오즈 결합 단백질(MBP)을 코딩하는 유전자와 서열번호 2로 표시되는 mSA 유전자 및 BBa_B0034 서열을 pBAD24에 다시 클로닝하여 pBAD_p2x-mSA(B_p-mSA), pBAD_c2x-mSA(B_c-mSA), pBAD_RBS1-p2x-mSA(B_R1.0-p-mSA), pBAD_RBS1-c2x-mSA(B_R1.0-c-mSA) 플라스미드를 제작하였다.
또한, pBAD-mSA 플라스미드를 주형으로 하여 mSA 유전자를 증폭한 뒤, 제한효소 EcoRⅠ 및 HindⅢ을 넣어 절단하고 이를 정제하여 유전자 증폭 산물을 얻은 뒤에 이를 동일한 제한효소로 절단한 pMAl_p2x, pMAl_c2x 플라스미드에 클로닝 하여 pMAl_p2x-mSA(M_p-mSA), pMAl_c2x-mSA(M_c-mSA) 플라스미드를 제작하였다.
[1-3] RBS 치환 플라스미드 제작
mSA 발현 수준 및 기능성을 증가시키기 위해서, 기존 RBS를 신규한 조절 유전자로 치환한 유전자 컨스트럭트를 추가로 제작하였다(하기 표 1 참고). 우선 플라스미드의 RBS 서열을 분석하여 서열 라이브러리를 제작하였다. 다음으로 RBS 계산기(Penn State University) 프로그램을 이용하여 B_p-mSA 플라스미드의 번역 개시 속도(TIR)를 분석한 후, RBS 라이브러리 계산기(RBS Library Calculator)를 통하여 번역 개시 속도의 값 범위가 3.97에서 42889인 조절 유전자 라이브러리를 구축하였다. 상기 라이브러리에 따라 제작된 조절 유전자를 B_p-mSA 플라스미드의 RBS 서열과 치환하여 클로닝한 후 생성된 콜로니를 선별하여 최종 플라스미드 pBAD_R01-p2x-mSA(B_R01-p-mSA), pBAD_R02-p2x-mSA(B_R02-p-mSA), pBAD_R1-p2x-mSA(B_R1-p-mSA), pBAD_R11-p2x-mSA(B_R11-p-mSA), pBAD_R12-p2x-mSA(B_R12-p-mSA), pBAD_R13-p2x-mSA(B_R13-p-mSA), pBAD_R2-p2x-mSA(B_R2-p-mSA), pBAD_R21-p2x-mSA(B_R21-p-mSA)를 제작하였다(하기 표 1 참고).
상기 실시예 [1-1] 내지 [1-3]에서 제작한 각 유전자 컨스트럭트의 명칭, 약자, 기본 플라스미드 및 전체 서열은 하기 표 1에 나타낸 바와 같다.
명칭
(약자)		 기본 플라스미드 유전자 컨스트럭트
mSA MBP RBS 전체 서열
pBAD-mSA
(B-mSA)		 pBAD24 서열번호 2 미추가 비치환 서열번호 8
pBAD_RBS 0.3-mSA
(B_R0.3-mSA)		 pBAD24 서열번호 2 미추가 서열번호 26 서열번호 9
pBAD_RBS 0.6-mSA
(B_R0.6-mSA)		 pBAD24 서열번호 2 미추가 서열번호 27 서열번호 10
pBAD_RBS 1.0-mSA
(B_R1.0-mSA)		 pBAD24 서열번호 2 미추가 서열번호 28 서열번호 11
pMAl_p2x-mSA
(M_p-mSA)		 pMAl_p2x 서열번호 2 서열번호 4 비치환 서열번호 12
pMAl_c2x-mSA
(M_c-mSA)		 pMAl_c2x 서열번호 2 서열번호 4 비치환 서열번호 13
pBAD_p2x-mSA
(B_p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 29 서열번호 14
pBAD_c2x-mSA
(B_c-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 29 서열번호 15
pBAD_RBS1-p2x-mSA
(B_R1.0-p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 28 서열번호 16
pBAD_RBS1-c2x-mSA
(B_R1.0-c-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 28 서열번호 17
pBAD_R01-p2x-mSA
(B_R01-p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 30 서열번호 18
pBAD_R02-p2x-mSA
(B_R02-p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 31 서열번호 19
pBAD_R1-p2x-mSA
(B_R1-p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 32 서열번호 20
pBAD_R11-p2x-mSA
(B_R11-p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 33 서열번호 21
pBAD_R12-p2x-mSA
(B_R12-p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 34 서열번호 22
pBAD_R13-p2x-mSA
(B_R13-p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 35 서열번호 23
pBAD_R2-p2x-mSA
(B_R2-p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 36 서열번호 24
pBAD_R21-p2x-mSA
(B_R21-p-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 37 서열번호 25
pBAD R-lib-1-1-mSA
(R-lib-1-1-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 65 서열번호 38
pBAD R-lib-1-5-mSA
(R-lib-1-5-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 66 서열번호 39
pBAD R-lib-1-7-mSA
(R-lib-1-7-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 67 서열번호 40
pBAD R-lib-1-10-mSA
(R-lib-1-10-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 68 서열번호 41
pBAD R-lib-1-11-mSA
(R-lib-1-11-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 69 서열번호 42
pBAD R-lib-1-12-mSA
(R-lib-1-12-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 70 서열번호 43
pBAD R-lib-1-13-mSA
(R-lib-1-13-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 71 서열번호 44
pBAD R-lib-1-14-mSA
(R-lib-1-14-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 72 서열번호 45
pBAD R-lib-1-16-mSA
(R-lib-1-16-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 73 서열번호 46
pBAD R-lib-1-17-mSA
(R-lib-1-17-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 74 서열번호 47
pBAD R-lib-1-18-mSA
(R-lib-1-18-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 75 서열번호 48
pBAD R-lib-2-2-mSA
(R-lib-2-2-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 76 서열번호 49
pBAD R-lib-2-3-mSA
(R-lib-2-3-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 77 서열번호 50
pBAD R-lib-2-4-mSA
(R-lib-2-4-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 78 서열번호 51
pBAD R-lib-2-5-mSA
(R-lib-2-5-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 79 서열번호 52
pBAD R-lib-2-6-mSA
(R-lib-2-6-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 80 서열번호 53
pBAD R-lib-2-7-mSA
(R-lib-2-7-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 81 서열번호 54
pBAD R-lib-2-8-mSA
(R-lib-2-8-mSA)		 pBAD24 서열번호 2 서열번호 4 서열번호 82 서열번호 55
pBAD R-lib-2-14-mSA
(R-lib-2-14-mSA		 pBAD24 서열번호 2 서열번호 4 서열번호 83 서열번호 56
pBAD R-lib-2-16-mSA
(R-lib-2-16-mSA		 pBAD24 서열번호 2 서열번호 4 서열번호 84 서열번호 57
pBAD R-lib-2-17-mSA
(R-lib-2-17-mSA		 pBAD24 서열번호 2 서열번호 4 서열번호 85 서열번호 58
pBAD R-lib-3-4-mSA
(R-lib-3-4-mSA		 pBAD24 서열번호 2 서열번호 4 서열번호 86 서열번호 59
pBAD R-lib-3-5-mSA
(R-lib-3-5-mSA		 pBAD24 서열번호 2 서열번호 4 서열번호 87 서열번호 60
pBAD R-lib-3-11-mSA
(R-lib-3-11-mSA		 pBAD24 서열번호 2 서열번호 4 서열번호 88 서열번호 61
pBAD R-lib-3-13-mSA
(R-lib-3-13-mSA		 pBAD24 서열번호 2 서열번호 4 서열번호 89 서열번호 62
pBAD R-lib-3-18-mSA
(R-lib-3-18-mSA		 pBAD24 서열번호 2 서열번호 4 서열번호 90 서열번호 63
pBAD R-lib-3-20-mSA
(R-lib-3-20-mSA		 pBAD24 서열번호 2 서열번호 4 서열번호 91 서열번호 64
[1-4] 조절 유전자 전사체의 총 깁스 자유 에너지 변화량 계산
상기 실시예 [1-3]에서 제작된 리보솜 결합 부위(RBS)의 프로모터로부터 개시코돈 까지의 서열에서, 총 깁스 자유 에너지 변화량(ΔGtotal)에 따른 상기 유전자 컨스트럭트의 mSA 발현 능력을 확인하기 위해, 조절 유전자의 mRNA와 리보솜의 30S 서브유닛의 복합체를 형성하는 반응이 일어날 때의 깁스 자유 에너지 변화량(ΔGmRNA-rRNA), 상기 조절 유전자의 mRNA 전사체에서 리보솜의 30S 서브유닛의 복합체를 형성하는 서열과 개시코돈 간의 간격(Spacing)이 최적화 되지 않음으로써 발생하는 대한 깁스 자유 에너지 패널티(ΔGspacing), 상기 간격(Spacing)의 영역에서 쌓인 뉴클레오티드의 깁스 자유 에너지 변화량(ΔGstacking), 상기 조절 유전자의 mRNA 전사체의 대기 부위(Stanby Site)와 리보솜 간 결합 반응이 일어나는 경우 깁스 자유 에너지 패널티(ΔGstandby), mRNA-tRNA 복합체를 형성하는 반응이 일어날 때의 깁스 자유 에너지 변화량(ΔGstart) 및 상기 조절 유전자의 mRNA 전사체가 접힌 복합체 구조물을 형성할 때의 깁스 자유 에너지 변화량(ΔGmRNA)을 계산하여 총 깁스 자유 에너지 변화량(ΔGtotal)을 계산한 결과를 하기 표 2에 나타내었다. 여기서, 상기 총 깁스 자유 에너지 변화량(ΔGtotal)은 하기 식 1 및 식 2와 같은 방법으로 계산되었다:
[식 1]
ΔGtotal = (ΔGfinal) - (ΔGinitial)
[식 2]
(ΔGfinal) - (ΔGinitial) = [(ΔGmRNA-rRNA) + (ΔGspacing) + (ΔGstacking) + (ΔGstandby) + (ΔGstart)] - (ΔGmRNA)
RBS 단위 kcal/mol
서열번호 ΔGmRNA-rRNA ΔGspacing ΔGstacking ΔGstandby ΔGstart ΔGmRNA ΔGtotal
29 -14.4914 4.992 0 2.41 -2.76 -15.01 4.944348676
30 -6.84135 1.525 0 1.5336 -2.76 -7.76 1.328848924
31 1.158649 0.005326 0 0.4314 -0.42 -3.4 4.641874373
32 -7.05135 0 0 1.0898 -2.76 -5.79 -3.555751381
33 -1.37135 0 0 0.0786 -2.76 -4.17 0.1498488
34 -0.24135 0 0 0.29 -2.76 -6.12 3.239348447
35 -5.39135 0.672 0 4.0728 -2.76 -7.41 3.862748471
36 -9.37135 0.288 0 4.0728 -2.76 -4.61 -3.458051381
37 -7.44135 0 0 4.8986 -2.76 -8.12 2.647948438
65 -7.44135 0 0 4.8986 -2.76 -10.25 4.777948552
66 -13.8614 0.288 0 4.0728 -2.76 -4.62 -7.9380514
67 -7.77135 0.288 0 4.0728 -2.76 -5.98 -0.488051352
68 -5.39135 0.288 0 4.0728 -2.76 -5.98 1.891948643
69 -2.68135 0.288 0 3.3234 -2.76 -4.61 2.482548676
70 -5.39135 0.288 0 4.0728 -2.76 -4.61 0.521948757
71 -2.68135 0.288 0 4.0728 -2.76 -5.37 3.991948428
72 -2.56135 0 0 3.3234 -2.76 -4.61 2.4427488
73 -3.96135 0.288 0 4.8986 -2.76 -6.51 4.67774886
74 -6.95135 0.288 0 4.8986 -2.76 -6.45 1.627748371
75 -7.49135 0.288 0 4.0728 -2.76 -6.78 0.59194881
76 -9.37135 0 0 2.6504 -2.76 -5.68 -4.216651686
77 -7.27135 0 0 2.6504 -2.76 -4.59 -3.283051219
78 -7.27135 0 0 2.6504 -2.76 -4.59 -2.966951219
79 -10.9714 0 0 2.6504 -2.76 -3.99 -7.41975141
80 -8.87135 0 0 2.6504 -2.76 -2.9 -6.510651419
81 -10.9714 0 0 2.6504 -2.76 -3.58 -7.860951495
82 -9.37135 0 0 2.6504 -2.76 -4.59 -5.403751362
83 -8.87135 0 0 2.6504 -2.76 -3.82 -5.729451581
84 -9.37135 0 0 2.6504 -2.76 -2.44 -7.456651457
85 -8.87135 0 0 2.6504 -2.76 -2.31 -7.100651572
86 -7.58135 1.525 0 0.0168 -2.76 -4.78 -4.109751104
87 -5.27135 0 0 0.4314 -2.76 -4.46 -3.219451333
88 -5.27135 0 0 0.4314 -2.76 -4.27 -3.76635139
89 -5.27135 0 0 0.4314 -2.76 -5.47 -2.476551581
90 -5.06135 0.005326 0 0.2168 -2.76 -4.1 -3.800725876
91 -7.75135 1.525 0 0.4314 -2.76 -4.95 -3.695151507
[1-5] 조절 유전자의 번역 개시 속도 계산
상기 조절 유전자의 번역 개시 속도(Translation Initiation Rate; TIR)에 따른 플라스미드의 mSA 발현 능력을 확인하기 위해, 상기와 같이 제작된 조절 유전자 서열에 따른 각 번역 개시 속도를 계산하여 하기 표 3와 같이 나타내었다.
조절 유전자 번역 개시 속도(au)
비치환된 경우 1
서열번호 29 133.266441
서열번호 30 678.2310709
서열번호 31 152.7003477
서열번호 32 6110.586323
서열번호 33 1152.963841
서열번호 34 287.0559877
서열번호 35 216.8312084
서열번호 36 5847.72872
서열번호 37 374.5906748
서열번호 65 143.6296318
서열번호 66 43914.95671
서열번호 67 1536.365645
서열번호 68 526.4033937
서열번호 69 403.5382313
서열번호 70 975.1871797
서열번호 71 204.582929
서열번호 72 410.8314232
서열번호 73 150.2547754
서열번호 74 592.8668512
서열번호 75 944.9445588
서열번호 76 8227.297678
서열번호 77 5404.842895
서열번호 78 4688.141139
서열번호 79 34778.40472
서열번호 80 23100.64943
서열번호 81 42417.31004
서열번호 82 14037.12899
서열번호 83 16253.13229
서열번호 84 35360.78091
서열번호 85 30125.96282
서열번호 86 7840.852282
서열번호 87 5252.334127
서열번호 88 6718.078741
서열번호 89 3759.681518
서열번호 90 6822.815917
서열번호 91 6506.222717
상기 표 3에서 나타낸 바와 같이, 제작된 플라스미드의 조절 유전자 중 서열번호 29 내지 37 및 65 내지 91의 조절 유전자는 번역 개시 속도가 50 내지 45000au 범위에 해당하였고, 그 중 서열번호 32 및 36의 조절 유전자는 번역 개시 속도가 900 내지 9000au 범위에 해당하는 것을 확인하였다.
[1-6] 조절 유전자의 서열 분석
상기 조절 유전자 서열의 AGG, TAGG 또는 ATAGG 서열 포함 여부 및 상기 AGG 서열의 3' 말단에서 개시코돈 까지의 간격(Spacing)에 따른 플라스미드의 mSA 발현 능력을 확인하기 위해, 각 플라스미드에 따른 조절 유전자 서열 및 상기 AGG 서열의 3' 말단에서 개시코돈 까지의 간격(Spacing, 단위 bp)을 하기 표 4와 같이 나타내었다.
조절 유전자 서열 Spacing(bp)
서열번호 26 TCACACAGGAAAG 4
서열번호 27 ATTAAAGAGGAGAAA 5
서열번호 28 AAAGAGGAGAAA 5
서열번호 29 ACCCGTTTTTTGGGCTAACAGGAGGAAGCTAGCGCTAGC 14
서열번호 30 TAGCACTCGTTGACATACGGACGTCAC -
서열번호 31 ACTACTGAGGCTACT 5
서열번호 32 TGGAACAGCTCACGCAAAAATAGGTTTCTT 6
서열번호 33 CGCTTTTTATCGCAACTCTCTACTGTTTCTCCAT -
서열번호 34 TCTGAGAAAGACACGATCTTACTAG -
서열번호 35 TCTAGAGAAAGAGCGGATCCTACCTAG -
서열번호 36 TCTAGAGAAAGATAGGAGAATACTAG 10
서열번호 37 TCTAGAGAAAGAGGCGACGGTACTAG 11
서열번호 65 TCTAGAGAAAGAGGCGAGTGTACTAG 12
서열번호 66 TCTAGAGAAAGATAGGAGGTTACTAG 10
서열번호 67 TCTAGAGAAAGAGGGGACACTACTAG 12
서열번호 68 TCTAGAGAAAGAGCGGAAACTACTAG -
서열번호 69 TTCTAGAGAAAGATTTGAATATACTAG -
서열번호 70 TCTAGAGAAAGAACGGACATTACTAG -
서열번호 71 TCTAGAGAAAGACATGACTATACTAG -
서열번호 72 TCTAGAGAAAGAACTGAAGATACTAG -
서열번호 73 TCTAGAGAAAGAGGCGATCCTACTAG 12
서열번호 74 TCTAGAGAAAGAAGAGAGCCTACTAG -
서열번호 75 TCTAGAGAAAGACTTGAGGCTACTAG 7
서열번호 76 GAACCCTAATACATTAGGAGATCTTCT 9
서열번호 77 GAACCCTAATACATTAGGACATATTCT 9
서열번호 78 GAACCCTAATACATTAGGACATCATCT 9
서열번호 79 GAACCCTAATACATAAGGAGATCATAT 9
서열번호 80 GAACCCTAATACATAAGGACATAATAT 9
서열번호 81 GAACCCTAATACATAAGGAGATTATCT 9
서열번호 82 GAACCCTAATACATTAGGAGATTATAT 9
서열번호 83 GAACCCTAATACATAAGGACATCTTAT 9
서열번호 84 GAACACTAATACATTAGGAGATCTTCT 9
서열번호 85 GAACACTAATACATAAGGACATAATAT 9
서열번호 86 TTAAGTAGTTAAACAGGGTATATAGGGGAAGA 6
서열번호 87 TTAAGTAGTTAAACAGGGTATATAGGACGAGA 6
서열번호 88 TTAAGTAGTTAAACAGGGTATATAGGGCTATA 6
서열번호 89 TTAAGTAGTTAAACAGGGTATATAGGAGGATA 6
서열번호 90 TTAAGTAGTTAAACAGGGTATATAGGGCGATA 6
서열번호 91 TTAAGTAATTAAACAGGGTATATAGGGGAAGA 6
상기 표 4에서 나타낸 바와 같이, 제작된 플라스미드의 조절 유전자 중 서열번호 26, 27, 28, 29, 31, 32, 36, 37, 65, 66, 67, 73, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90 및 91의 조절 유전자는 AGG 서열을 포함하고 있었고, 그 중 서열번호 32 및 36의 조절 유전자는 상기 AGG 서열의 3' 말단에서 개시코돈 까지의 간격(Spacing)이 6 내지 13bp인 것을 확인하였다. 또한 제작된 플라스미드의 조절 유전자 중 서열번호 32,36, 66, 76, 77, 78, 82, 84, 86, 87, 88, 89, 90 및 91의 조절 유전자는 TAGG 또는 ATAGG 서열을 포함하는 것을 확인하였다.
[실시예 2] 숙주 세포의 형질 전환 및 배양
상기 실시예 1에서 제작한 플라스미드를 박테리아 균주에 형질 전환한 뒤, 엠피실린(Ampicillin)이 포함된 LB 고체 배지를 이용하여 상기 형질 전환된 각각의 균주를 밤새 배양하였다. 그 뒤, 생성된 콜로니를 새로 항생제가 포함된 LB 액체 배지를 사용하여 1:100의 비율로 희석하고, 추가 배양을 통해 OD600 값이 0.5 내지 0.7에 도달할 때, 상기 배양액에 최종 농도가 0.1%인 아라비노오스(arabinose)를 첨가하고 200rpm 및 37℃의 조건으로 진탕 배양기에서 배양하였다.
[실험예 1] 재조합 mSA 플라스미드의 mSA 발현 수준 확인
mSA 유전자를 단독으로 삽입한 플라스미드의 발현 수준을 확인하기 위해 상기 실시예 1에서 제작된 플라스미드 B-mSA, B_R0.3-mSA, B_R0.6-mSA, B_R1.0-mSA를 포함하는 재조합 대장균 콜로니를 실시예 2와 같이 형질 전환 및 배양한 후, 배양된 재조합 대장균을 OD4 기준으로 SDS-PAGE 샘플 버퍼를 첨가하여 95도에서 10분간 끓인 후 SDS-PAGE에 로딩하여 단백질 발현 정도를 확인하여 도 1에 나타내었다.
도 1에서 나타난 바와 같이, 단백질 발현을 개선하기 위해 BBa_B0032, BBa_B0030, BBa_B0034로 알려진 RBS 서열을 mSA 유전자 서열 앞에 삽입한 플라스미드를 포함한 균주들은 SDS-PAGE 상에서 mSA 단백질 발현을 확인할 수 없었다. 이를 통해서 mSA 유전자를 단독 발현함에 있어서 pBAD 발현 시스템에서는 RBS 서열의 추가는 유전자가 과발현에 영향을 크게 미치지 않음을 알 수 있었다.
[실험예 2] MBP-mSA 플라스미드의 mSA 발현 수준 및 활성 확인
[2-1] SDS-PAGE
mSA의 발현과 가용성을 증대시키기 위해 MBP 유전자를 융합한 MBP-mSA 플라스미드로 형질 전환된 균주의 mSA 발현 수준을 확인하였다. 구체적으로는, MBP 유전자를 융합한 M_p-mSA, M_c-mSA 플라스미드를 상기 실시예 1과 같이 제작한 후, 상기 실시예 2와 같이 형질 전환 및 배양하였다. 배양 중 OD600 값이 0.5 내지 0.7에 도달할 때 상기 배양액에 최종 농도가 0.1 mM인 IPTG(Isopropyl beta-D-1-thiogalactopyranoside)를 첨가하고 200 rpm 및 37℃의 조건으로 진탕 배양기에서 배양하였다. 배양된 재조합 대장균은 OD4 기준으로 SDS-PAGE sample buffer를 첨가하여 95도에서 10분간 끓인 후 SDS-PAGE 로딩 및 단백질 발현 수준을 확인하여 도 2에 나타내었다.
그 결과, 도 2와 같이, MBP와 융합한 mSA 단백질은 겔(gel) 상에서 과발현됨을 확인하였으며 분비 서열이 없는 MBP 유전자를 융합한 mSA 유전자가 분비 서열이 있는 경우 보다 더 과발현됨을 알 수 있었다.
[2-2] 웨스턴 블롯 분석
MBP-mSA 플라스미드로 형질 전환된 재조합 균주의 mSA 발현 수준과 비오틴 결합 활성을 확인하기 위해 웨스턴 블롯 분석을 수행하였다. 구체적으로, 상기 실시예 2의 균주의 배양액을 4 x 107 CFU/ml이 되도록 PBS를 이용하여 희석하고, 13,000rpm에서 5분동안 원심분리 하여 펠렛을 수집하였다. 상기 펠렛 분획을 PBS를 이용하여 세척하고, 0.2% 베타-머캅토에탄올(beta-mercaptoethanol)(카탈로그 번호: EBA-1052, ELPIS BIOTECH)이 포함된 SDS 샘플 완충액과 혼합하여 균주 용해물을 수득하였다. 그런 다음, 12% SDS-PAGE에서 상기 균주 용해물을 전기영동 하고, 상기 겔로부터 니트로 셀룰로스 막에 단백질을 옮긴 뒤 5% 탈지유를 이용하여 상온에서 블로킹 하였다. 이후, his tag 항체를 이용하여 mSA의 발현 수준을 확인하고, 비오틴화된 과산화효소(Biotinylated peroxidase)를 이용하여 mSA의 비오틴 결합 활성을 확인하였다. 수행하여 도 3에 나타내었다.
도 3에서 나타난 바와 같이, MBP 유전자와 mSA 유전자를 모두 삽입한 M_c-mSA 및 M_p-mSA 플라스미드의 MBP와 융합한 mSA 단백질 발현량이 단순 mSA 유전자만 삽입한 B-mSA 플라스미드의 MBP와 융합하지 않은 단순 mSA 단백질 발현량 보다 높았다.
또한, M_c-mSA 플라스미드에서 발현된 단백질이 M_p-mSA 보다 많음에도 불구하고 biotin과의 결합 활성은 분비 서열이 있는 MBP와 융합한 mSA가 더 높음을 확인할 수 있었다.
[2-3] 비오틴 흡수 분석
MBP-mSA 플라스미드로 형질 전환된 재조합 균주의 비오틴 결합 활성을 분석하기 위해 비오틴 흡수 분석(biotin uptake assay)을 수행하여 그 결과를 도 4와 같이 나타내었다. 구체적으로, 배양된 균주에 비오틴화된 형광염료(biotinylated fluorescent dye, biotin-flamma 675 dye, BioActs)를 첨가하여 반응한 후, PBS로 세척하여 균주에 결합하지 않는 비오틴화된 형광염료를 제거하여 형광 측정 리더기(Infinite m200, Tecan)를 이용하여 재조합 균주에 흡수된 형광염료의 형광값을 측정하였다.
그 결과, 도 4와 같이, 아라비노즈 첨가 전/후 비오틴 결합 시그널 증가량이 대조군인 B-mSA 플라스미드를 포함하는 균주(344%)가 pBAD(151%), M_c-mSA(141%) 및 M_p-mSA(158%)를 포함하는 균주보다 비오틴 활성이 더 높아 상기 MBP-mSA 발현 재조합 균주의 비오틴 결합능이 개선되지 않음을 확인하였다.
[2-4] 공초점 현미경 관찰
실제 재조합 균주와 비오틴화된 형광염료의 결합을 이미지로 확인하기 위해 상기 실시예 2와 같이 배양한 재조합 균주를 슬라이드에 고정하여 공초점 현미경으로 확인하여 도 5 및 도 6과 같이 나타내었다.
도 5 및 도 6의 비오틴 흡수 분석(biotin uptake assay)의 결과와 같이 MBP 융합 여부와 상관없이 균주에 결합한 비오틴화된 형광염료는 매우 적은 수를 보였다. 이를 통해 MBP 유전자 융합을 통해 mSA의 발현이 개선되었으나, MBP를 통한 유전자의 발현 및 가용성의 변화가 비오틴 결합 활성을 개선시킬 수 없다는 것을 확인할 수 있었다.
[실험예 3] RBS 추가 플라스미드의 mSA 발현 수준 및 활성 확인
[3-1] SDS-PAGE
RBS가 추가된 플라스미드로 형질 전환된 재조합 균주의 mSA의 발현 및 활성을 확인하기 위해 SDS-PAGE를 수행하였다. 구체적으로는, MBP-mSA 유전자를 pBAD 플라스미드에 옮긴 B_p-mSA, B_c-mSA 및 상기 플라스미드의 발현을 개선하기 위해 BBa_B0034 서열을 추가한 B_R1.0-p-mSA, B_R1.0-c-mSA 플라스미드로 형질 전환된 재조합 균주에 대하여 SDS-PAGE를 수행한 결과를 도 7과 같이 나타내었다.
도 7에서 나타난 바와 같이, 상기 재조합 균주를 SDS-PAGE에 로딩하여 단백질 발현 정도를 확인한 결과, pBAD 플라스미드에서도 MBP와 융합한 mSA 단백질은 과발현됨을 확인하였다.
[3-2] 웨스턴 블롯 분석
RBS가 추가된 플라스미드로 형질 전환된 재조합 균주의 mSA의 발현 및 활성을 확인하기 위해 웨스턴 블롯 분석을 수행하였다. MBP-mSA 유전자를 pBAD 플라스미드에 옮긴 B_p-mSA, B_c-mSA와 이 플라스미드의 발현을 개선하기 위해 BBa_B0034 서열을 추가한 B_R1.0-p-mSA, B_R1.0-c-mSA 플라스미드에 대하여 상기 실험예 2-2와 같은 방식으로 웨스턴 블롯 분석을 수행하여 도 8과 같이 나타내었다.
도 8에서 나타난 바와 같이, 웨스턴 블롯 분석을 수행한 결과, MBP 유전자와 mSA 유전자를 모두 삽입한 B_p-mSA, B_c-mSA, B_R1.0-p-mSA 및 B_R1.0-c-mSA 플라스미드는 MBP와 융합한 mSA 단백질을 과발현하는 것을 확인하였으며, 이는 MBP와 융합하지 않은 단순 mSA 단백질 발현량 보다 높았다.
[실험예 4] RBS 치환 플라스미드의 mSA 발현 수준 및 활성 확인
[4-1] 웨스턴 블롯 분석(1)
본 발명자는 재조합 균주에서 유전자의 기능성 발현이 증가하게 유도되도록 하기 위해서 B_p-mSA 플라스미드의 RBS 서열을 분석하여 상기 실시예 1과 같이 B_R01-p-mSA, B_R02-p-mSA, B_R1-p-mSA, B_R11-p-mSA, B_R12-p-mSA, B_R13-p-mSA B_R2-p-mSA, B_R21-p-mSA 플라스미드를 제작한 후 균주에 형질 전환 및 배양하였다. 상기 재조합 균주의 단백질 발현 정도를 확인하기 위해 상기 실험예 2-2와 같은 방식으로 웨스턴 블롯 분석을 수행하여 도 9와 같이 나타내었다.
도 9에서 나타난 바와 같이, mSA를 발현하는 균주 중 B_R1-p-mSA와 B_R2-p-mSA 플라스미드로 형질 전환된 재조합 균주가 다른 균주에 비해 mSA 발현 수준이 뛰어난 것을 확인하였다.
[4-2] 웨스턴 블롯 분석(2)
mSA 발현 수준이 높은 B_R1-p-mSA와 B_R2-p-mSA 플라스미드로 형질 전환된 상기 두 균주를 선정하여 추가 실험을 수행하여, 그 결과를 도 10에 나타내었다.
도 10에서 나타난 바와 같이, 대조군에서는 M_p-mSA 플라스미드를 포함하는 재조합 균주가 B_p-mSA 플라스미드를 포함하는 재조합 균주보다 mSA의 발현과 분비 수준이 높았다. 실험군에서도 M_p-mSA 플라스미드를 포함하는 재조합 균주가 B_R1-p-mSA와 B_R2-p-mSA 플라스미드를 포함하는 재조합 균주 보다 mSA의 발현과 분비 수준이 높음을 확인하였다.
또한, B_R1-p-mSA와 B_R2-p-mSA 플라스미드를 포함하는 재조합 균주가 M_p-mSA 플라스미드를 포함하는 재조합 균주보다 발현양 대비 분비된 단백질은 더 적은 것으로 나타나서, 상기 B_R1-p-mSA와 B_R2-p-mSA 플라스미드에서 발현된 mSA는 균주의 주변세포질(periplasm)에 머무는 것을 알 수 있었다. 비오틴 결합 활성은 B_p-mSA, B_R1-p-mSA, M_p-mSA, B_R2-p-mSA 플라스미드를 포함하는 재조합 균주 순이였으며, 분비된 단백질 결합 활성은 M_p-mSA, B_p-mSA, B_R1-p-mSA, B_R2-p-mSA 플라스미드를 포함하는 재조합 균주 순으로 확인되었다.
[4-3] 비오틴 흡수 분석
또한, 발현이 개선된 재조합 균주의 비오틴 결합 활성을 분석하기 위해 실험예 2와 같은 방식으로 비오틴 흡수 분석(biotin uptake assay)을 수행하여 그 결과를 도 11과 같이 나타내었다.
*도 11에서 나타난 바와 같이, 대조군으로 pBAD와 B-mSA 플라스미드를 포함하는 재조합 균주보다, B_R1-p-mSA와 B_R2-p-mSA 플라스미드를 포함하는 재조합 균주가 비오틴 결합 활성이 현저히 높아, 상기 B_R1-p-mSA와 B_R2-p-mSA 플라스미드의 유전자에서 발현된 mSA가 다른 플라스미드에서 발현된 mSA에 비하여 현저한 비오틴 결합 활성 효과를 가지는 것을 확인하였다. 또한, 비오틴 결합 활성은 단백질 발현량과는 비례하지 않아, 단순히 단백질 발현량 만으로는 비오틴 결합 활성 효과를 예측할 수 없음을 확인하였다.
[4-4] 공초점 현미경 관찰
실제 재조합 균주와 비오틴화된 형광염료의 결합을 이미지로 확인하기 위해 상기 배양한 균주를 슬라이드에 고정하여 도 12 및 도 13과 같이 공초점 현미경으로 확인하였다.
도 12에서 나타난 바와 같이, B_R1-p-mSA와 B_R2-p-mSA 플라스미드를 포함하는 재조합 균주는 도 13에 나타난 대조군인 B-mSA 및 B_p-mSA에 비하여 비오틴화된 형광염료가 균주에 강하게 결합하는 것을 확인할 수 있었으며, 특히, B_R2-p-mSA 플라스미드에 의한 mSA 발현이 비오틴화된 형광염료와 결합하기에 최적의 상태임을 알 수 있었다. 이를 통해 MBP 유전자 융합을 통해 mSA 유전자의 발현이 개선된 균주의 경우에도 외부 비오틴과의 충분한 결합을 하지 못하였으나, mSA 유전자와 함께 MBP 유전자 및 RBS 유전자를 결합시킨 경우 mSA 유전자의 발현이 보다 기능적으로 개선되어 외부 비오틴과의 결합능이 매우 향상됨을 알 수 있었다.
[실험예 5] mSA 발현 재조합 균주에 대한 추적 기능 확인
[5-1] 비오틴 흡수 분석
상기 제작된 본 발명의 재조합 균주에서 발현된 mSA 유전자가 비오틴(biotin)에 특이성을 갖는지 확인하기 위해 대장균(E. coli) 균주에 pBAD, B-mSA, B_p-mSA, B_R1-p-mSA, B_R2-p-mSA 플라스미드를 실험예 1과 같이 형질 전환 및 배양 후, 상기 실험예 2와 같은 방식으로 비오틴 흡수 분석(biotin uptake assay)을 수행하여 그 결과를 도 14 및 도 15에 나타내었다.
도 14에서 나타난 바와 같이, 비오틴화된 형광염료만 처리하였을 때 B_R2-p-mSA 플라스미드를 포함하는 재조합 대장균(E. coli) 균주가 가장 높은 비오틴 흡수(biotin uptake)를 보였다. 반면, 도 15에서 나타난 바와 같이, 형광염료가 없는 비오틴을 미리 첨가한 후 비오틴화된 형광염료를 처리하였을 때 재조합 균주의 비오틴화된 형광염료의 흡수가 감소함을 확인하였다. 상기 도 14와 도 15 간의 차이를 통하여 비오틴화된 형광염료를 첨가하기 전에 형광염료가 없는 200 nM의 비오틴을 첨가하는 경우 비오틴화된 형광염료와 재조합 균주의 결합 저해할 수 있다는 것을 확인하였고, 이를 통하여 본 발명의 재조합 균주는 비오틴에 특이적으로 결합함을 알 수 있었다.
[5-2] 근육조직에서 균주 추적(1)
본 발명에 따른 근육조직에서 균주 추적 효과를 확인하기 위해 in vivo 이미지 시스템(In Vivo Imaging System; IVIS) 촬영을 실시하였다. 구체적으로는, 마우스(BALB/C)에 B_R2-p-mSA에 의해 형질 전환된 재조합 균주 1x109 CFU를 오른쪽 허벅지 근육내 주사(Intramuscle injection; IM)하였다. 이후 아라비노스(arabinose)를 주입하여 상기 재조합 균주에 mSA를 발현하였고, 대조군에서는 아라비노스를 주입하지 않았다. 이후 실험군과 대조군 각각에 비오틴 염색(Biotin-dye)을 주입하여 시그널을 확인한 결과를 도 16에 나타내었다.
도 16에 나타난 것과 같이, 상기 재조합 균주와 아라비노스를 주입한 마우스(오른쪽)의 오른쪽 허벅지에는 6시간이 넘게 비오틴 염색에 의한 강한 시그널이 발생하였으나, 아라비노스를 주입하지 않은 대조군 마우스(왼쪽)의 경우, 비오틴 염색에 의한 시그널이 없는 것을 확인한 바, 상기 재조합 균주와 아라비노스를 주입하여 mSA를 발현시킨 후 비오틴 염색을 이용할 경우, 근육조직에서 상기 재조합 균주의 분포를 추적할 수 있음을 확인하였다.
[5-3] 근육조직에서 균주 추적(2)
본 발명의 재조합 균주가 실제 근육조직에 분포하는지 확인하기 위해 앞서 실험예 [5-2]에서 균주를 주입했던 오른쪽 허벅지 조직을 적출하여 균주의 개체수를 확인하였다. 구체적으로는 실험예 [5-2]에서 비오틴 염색에 의한 재조합 균주의 시그널이 존재하는 마우스(Induction)와 대조군 마우스(Non-induction) 각각에 대한 오른쪽 허벅지 조직을 적출하여, 잔류하는 균주의 개체수를 확인한 결과를 도 17에 나타내었다.
도 17에 나타난 것과 같이, 본 발명의 재조합 균주는 아라비노스 처리 유무와 상관없이 허벅지 근육조직 모두에 균주가 존재하는 것으로 확인되는 바, 근육조직에서 상기 재조합 균주의 분포는 상기 아라비노스 처리에 의해 재조합 균주에서 발현된 mSA에 의해 추적할 수 있는 것임을 확인하였다.
[5-4] 복강으로 투여한 균주의 추적(1)
본 발명에 따른 균주 추적 효과를 확인하기 위해 복강으로 주입한 균주에 대한 in vivo 이미지 시스템(In Vivo Imaging System; IVIS) 촬영을 실시하였다. 구체적으로는, 마우스(BALB/C)에 B_R2-p-mSA에 의해 형질 전환된 재조합 균주 5x109 CFU를 마우스 복강내 주사 (Intraperitonealy injection; IP)하였다. 이후 아라비노스(arabinose)를 주입하여 상기 재조합 균주에 mSA를 발현하였고, 대조군에서는 아라비노스를 주입하지 않았다. 이후 실험군과 대조군 각각에 비오틴 염색(Biotin-dye)을 주입하여 시그널을 확인한 결과를 도 18에 나타내었다.
도 18에 나타난 것과 같이, 상기 재조합 균주와 아라비노스를 주입한 마우스(오른쪽)의 복강 장기에서는 6시간이 넘게 비오틴 염색에 의한 강한 시그널이 발생하였으나, 재조합 균주를 주입하지 않은 대조군(왼쪽) 또는 아라비노스를 주입하지 않은 대조군 마우스(가운데)의 경우, 6시간이 지났을 때 비오틴 염색에 의한 시그널이 사라진 것을 확인하였고, 장을 적출하여 시그널을 확인한 결과에서도 상기 재조합 균주와 아라비노스를 주입한 마우스(오른쪽)에서만 시그널이 발생한 것을 확인한 바, 상기 재조합 균주와 아라비노스를 주입하여 mSA를 발현시킨 후 비오틴 염색을 이용할 경우, 복강에 주입한 상기 재조합 균주의 분포를 추적할 수 있음을 확인하였다.
[5-5] 복강으로 투여한 균주의 추적(2)
본 발명의 재조합 균주가 실제 복강 및 장관에 분포하는지 확인하기 위해 앞서 실험예 [5-4]에서 적출한 장에 대하여 균주의 개체수를 확인하였다. 구체적으로는 실험예 [5-4]에서 비오틴 염색에 의한 재조합 균주의 시그널이 존재하는 마우스(Induction)와 대조군 마우스(Non-induction) 각각에 대한 장을 적출하여, 잔류하는 균주의 개체수를 확인한 결과를 도 19에 나타내었다.
도 19에 나타난 것과 같이, 본 발명의 재조합 균주는 아라비노스 처리 유무와 상관없이 장관에 균주가 존재하는 것으로 확인되는 바, 복강 및 장관에서 상기 재조합 균주의 분포는 상기 아라비노스 처리에 의해 재조합 균주에서 발현된 mSA에 의해 추적할 수 있는 것임을 확인하였다.
[5-6] 정맥으로 투여한 균주의 추적(1)
본 발명에 따른 균주 추적 효과를 확인하기 위해 정맥으로 주입한 균주에 대한 in vivo 이미지 시스템(In Vivo Imaging System; IVIS) 촬영을 실시하였다. 구체적으로는, 마우스(BALB/C)에 B_R2-p-mSA에 의해 형질 전환된 재조합 균주 1x109 CFU를 마우스 정맥내 주사(Intravenously injection; IV)하였다. 이후 아라비노스(arabinose)를 주입하여 상기 재조합 균주에 mSA를 발현하였고, 대조군에서는 아라비노스를 주입하지 않았다. 이후 실험군과 대조군 각각에 비오틴 염색(Biotin-dye)을 주입하여 시그널을 확인한 결과를 도 20에 나타내었다.
도 20에 나타난 것과 같이, 상기 재조합 균주와 아라비노스를 주입한 마우스(오른쪽)에는 6시간이 넘게 비오틴 염색에 의한 강한 시그널이 발생하였으나, 재조합 균주를 주입하지 않은 대조군(왼쪽) 또는 아라비노스를 주입하지 않은 대조군 마우스(가운데)의 경우, 6시간이 지났을 때 비오틴 염색에 의한 시그널이 사라진 것을 확인하였고, 간과 비장을 적출하여 시그널을 확인한 결과에서도 상기 재조합 균주와 아라비노스를 주입한 마우스(오른쪽)에서만 시그널이 발생한 것을 확인한 바, 상기 재조합 균주와 아라비노스를 주입하여 mSA를 발현시킨 후 비오틴 염색을 이용할 경우, 장기에서 상기 재조합 균주의 분포를 추적할 수 있음을 확인하였다.
[5-7] 정맥으로 투여한 균주의 추적(2)
본 발명의 재조합 균주가 실제 장기에 분포하는지 확인하기 위해 앞서 실험예 [5-6]에서 적출한 간에 대하여 균주의 개체수를 확인하였다. 구체적으로는 실험예 [5-6]에서 비오틴 염색에 의한 재조합 균주의 시그널이 존재하는 마우스(Induction)와 대조군 마우스(Non-induction) 각각에 대한 간 및 비장을 적출하여, 잔류하는 균주의 개체수를 확인한 결과를 도 21에 나타내었다.
도 21에 나타난 것과 같이, 본 발명의 재조합 균주는 아라비노스 처리 유무와 상관없이 간에 균주가 존재하는 것으로 확인되는 바, 생체 내에서 상기 재조합 균주의 분포는 상기 아라비노스 처리에 의해 재조합 균주에서 발현된 mSA에 의해 추적할 수 있는 것임을 확인하였다.
[5-8] 경구 투여한 균주의 추적(1)
본 발명에 따른 균주 추적 효과를 확인하기 위해 경구 투여한 균주에 대한 in vivo 이미지 시스템(In Vivo Imaging System; IVIS) 촬영을 실시하였다. 구체적으로는, 마우스(BALB/C)에 B_R2-p-mSA에 의해 형질 전환된 재조합 균주 1x109 CFU를 경구 투여(Oral administration) 하였다. 이후 아라비노스(arabinose)를 주입하여 상기 재조합 균주에 mSA를 발현하였고, 대조군에서는 아라비노스를 주입하지 않았다. 이후 실험군과 대조군 각각에 비오틴 염색(Biotin-dye)을 주입하여 시그널을 확인한 결과를 도 22에 나타내었다.
도 22에 나타난 것과 같이, 상기 재조합 균주와 아라비노스를 주입한 마우스(오른쪽)에는 6시간이 넘게 장관에서 비오틴 염색에 의한 강한 시그널이 발생하였으나, 재조합 균주를 주입하지 않은 대조군(왼쪽) 또는 아라비노스를 주입하지 않은 대조군 마우스(가운데)의 경우, 6시간이 지났을 때 비오틴 염색에 의한 시그널이 사라진 것을 확인하였고, 장을 적출하여 시그널을 확인한 결과에서도 상기 재조합 균주와 아라비노스를 주입한 마우스(오른쪽)에서만 시그널이 발생한 것을 확인한 바, 상기 재조합 균주와 아라비노스를 주입하여 mSA를 발현시킨 후 비오틴 염색을 이용할 경우, 장관에서 상기 재조합 균주의 분포를 추적할 수 있음을 확인하였다.
[5-9] 경구 투여한 균주의 추적(2)
본 발명의 재조합 균주가 실제 장관에 분포하는지 확인하기 위해 앞서 실험예 [5-8]에서 적출한 장에 대하여 균주의 개체수를 확인하였다. 구체적으로는 실험예 [5-8]에서 비오틴 염색에 의한 재조합 균주의 시그널이 존재하는 마우스(Induction)와 대조군 마우스(Non-induction) 각각에 대한 장을 적출하여, 잔류하는 균주의 개체수를 확인한 결과를 도 23
에 나타내었다.
도 23에 나타난 것과 같이, 본 발명의 재조합 균주는 아라비노스 처리 유무와 상관없이 장관에 균주가 존재하는 것으로 확인되는 바, 장관에서 상기 재조합 균주의 분포는 상기 아라비노스 처리에 의해 재조합 균주에서 발현된 mSA에 의해 추적할 수 있는 것임을 확인하였다.
구체적으로 상기 실험을 통해, 본 발명에 따르는 재조합 벡터 내지 컨스트럭트, 특히는 본 발명에 따르는 조절 유전자를 포함하는 경우 발현되는 모노머형 스트렙타비딘(mSA)의 안정성이 뛰어나며 외부 비오틴과 강하게 결합할 수 있고, 이는 생체 내에서도 유효하며, 비오틴화된 형광염료를 다회 처리하거나 시간을 조절하여 처리할 수도 있음을 확인하였다.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현 예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.
<110> Industry Foundation of Channam National University <120> In vivo microbial tracking platform comprising a strain expressing pre-targeting protein <130> PDPB204242k01 <150> KR 10-2021-0006608 <151> 2021-01-18 <160> 91 <170> KoPatentIn 3.0 <210> 1 <211> 115 <212> PRT <213> Artificial Sequence <220> <223> mSA protein <400> 1 Met Ala Glu Ala Gly Ile Thr Gly Thr Trp Tyr Asn Gln His Gly Ser 1 5 10 15 Thr Phe Thr Val Thr Ala Gly Ala Asp Gly Asn Leu Thr Gly Gln Tyr 20 25 30 Glu Asn Arg Ala Gln Gly Thr Gly Cys Gln Asn Ser Pro Tyr Thr Leu 35 40 45 Thr Gly Arg Tyr Asn Gly Thr Lys Leu Glu Trp Arg Val Glu Trp Asn 50 55 60 Asn Ser Thr Glu Asn Cys His Ser Arg Thr Glu Trp Arg Gly Gln Tyr 65 70 75 80 Gln Gly Gly Ala Glu Ala Arg Ile Asn Thr Gln Trp Asn Leu Thr Tyr 85 90 95 Glu Gly Gly Ser Gly Pro Ala Thr Glu Gln Gly Gln Asp Thr Phe Thr 100 105 110 Lys Val Lys 115 <210> 2 <211> 348 <212> DNA <213> Artificial Sequence <220> <223> mSA gene <400> 2 atggcggaag cgggtatcac cggcacgtgg tacaaccagc atggttctac cttcaccgtt 60 accgcgggtg cggacggtaa cctgaccggt cagtacgaaa accgtgcgca gggcactggt 120 tgccagaact ctccgtacac cctgaccggt cgttacaacg gtaccaaact ggaatggcgt 180 gttgaatgga acaactctac cgaaaactgc cactctcgta ccgaatggcg tggtcagtac 240 cagggtggtg cggaagcgcg tatcaacacc cagtggaacc tgacctacga aggtggttct 300 ggtccggcga ccgaacaggg tcaggacacc ttcaccaaag ttaaataa 348 <210> 3 <211> 392 <212> PRT <213> Artificial Sequence <220> <223> MBP protein <400> 3 Met Lys Ile Lys Thr Gly Ala Arg Ile Leu Ala Leu Ser Ala Leu Thr 1 5 10 15 Thr Met Met Phe Ser Ala Ser Ala Leu Ala Lys Ile Glu Glu Gly Lys 20 25 30 Leu Val Ile Trp Ile Asn Gly Asp Lys Gly Tyr Asn Gly Leu Ala Glu 35 40 45 Val Gly Lys Lys Phe Glu Lys Asp Thr Gly Ile Lys Val Thr Val Glu 50 55 60 His Pro Asp Lys Leu Glu Glu Lys Phe Pro Gln Val Ala Ala Thr Gly 65 70 75 80 Asp Gly Pro Asp Ile Ile Phe Trp Ala His Asp Arg Phe Gly Gly Tyr 85 90 95 Ala Gln Ser Gly Leu Leu Ala Glu Ile Thr Pro Asp Lys Ala Phe Gln 100 105 110 Asp Lys Leu Tyr Pro Phe Thr Trp Asp Ala Val Arg Tyr Asn Gly Lys 115 120 125 Leu Ile Ala Tyr Pro Ile Ala Val Glu Ala Leu Ser Leu Ile Tyr Asn 130 135 140 Lys Asp Leu Leu Pro Asn Pro Pro Lys Thr Trp Glu Glu Ile Pro Ala 145 150 155 160 Leu Asp Lys Glu Leu Lys Ala Lys Gly Lys Ser Ala Leu Met Phe Asn 165 170 175 Leu Gln Glu Pro Tyr Phe Thr Trp Pro Leu Ile Ala Ala Asp Gly Gly 180 185 190 Tyr Ala Phe Lys Tyr Glu Asn Gly Lys Tyr Asp Ile Lys Asp Val Gly 195 200 205 Val Asp Asn Ala Gly Ala Lys Ala Gly Leu Thr Phe Leu Val Asp Leu 210 215 220 Ile Lys Asn Lys His Met Asn Ala Asp Thr Asp Tyr Ser Ile Ala Glu 225 230 235 240 Ala Ala Phe Asn Lys Gly Glu Thr Ala Met Thr Ile Asn Gly Pro Trp 245 250 255 Ala Trp Ser Asn Ile Asp Thr Ser Lys Val Asn Tyr Gly Val Thr Val 260 265 270 Leu Pro Thr Phe Lys Gly Gln Pro Ser Lys Pro Phe Val Gly Val Leu 275 280 285 Ser Ala Gly Ile Asn Ala Ala Ser Pro Asn Lys Glu Leu Ala Lys Glu 290 295 300 Phe Leu Glu Asn Tyr Leu Leu Thr Asp Glu Gly Leu Glu Ala Val Asn 305 310 315 320 Lys Asp Lys Pro Leu Gly Ala Val Ala Leu Lys Ser Tyr Glu Glu Glu 325 330 335 Leu Ala Lys Asp Pro Arg Ile Ala Ala Thr Met Glu Asn Ala Gln Lys 340 345 350 Gly Glu Ile Met Pro Asn Ile Pro Gln Met Ser Ala Phe Trp Tyr Ala 355 360 365 Val Arg Thr Ala Val Ile Asn Ala Ala Ser Gly Arg Gln Thr Val Asp 370 375 380 Glu Ala Leu Lys Asp Ala Gln Thr 385 390 <210> 4 <211> 1179 <212> DNA <213> Artificial Sequence <220> <223> MBP gene <400> 4 atgaaaataa aaacaggtgc acgcatcctc gcattatccg cattaacgac gatgatgttt 60 tccgcctcgg ctctcgccaa aatcgaagaa ggtaaactgg taatctggat taacggcgat 120 aaaggctata acggtctcgc tgaagtcggt aagaaattcg agaaagatac cggaattaaa 180 gtcaccgttg agcatccgga taaactggaa gagaaattcc cacaggttgc ggcaactggc 240 gatggccctg acattatctt ctgggcacac gaccgctttg gtggctacgc tcaatctggc 300 ctgttggctg aaatcacccc ggacaaagcg ttccaggaca agctgtatcc gtttacctgg 360 gatgccgtac gttacaacgg caagctgatt gcttacccga tcgctgttga agcgttatcg 420 ctgatttata acaaagatct gctgccgaac ccgccaaaaa cctgggaaga gatcccggcg 480 ctggataaag aactgaaagc gaaaggtaag agcgcgctga tgttcaacct gcaagaaccg 540 tacttcacct ggccgctgat tgctgctgac gggggttatg cgttcaagta tgaaaacggc 600 aagtacgaca ttaaagacgt gggcgtggat aacgctggcg cgaaagcggg tctgaccttc 660 ctggttgacc tgattaaaaa caaacacatg aatgcagaca ccgattactc catcgcagaa 720 gctgccttta ataaaggcga aacagcgatg accatcaacg gcccgtgggc atggtccaac 780 atcgacacca gcaaagtgaa ttatggtgta acggtactgc cgaccttcaa gggtcaacca 840 tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg ccgccagtcc gaacaaagag 900 ctggcaaaag agttcctcga aaactatctg ctgactgatg aaggtctgga agcggttaat 960 aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg aggaagagtt ggcgaaagat 1020 ccacgtattg ccgccactat ggaaaacgcc cagaaaggtg aaatcatgcc gaacatcccg 1080 cagatgtccg ctttctggta tgccgtgcgt actgcggtga tcaacgccgc cagcggtcgt 1140 cagactgtcg atgaagccct gaaagacgcg cagacttaa 1179 <210> 5 <211> 3 <212> DNA <213> Artificial Sequence <220> <223> DNA site <400> 5 agg 3 <210> 6 <211> 4 <212> DNA <213> Artificial Sequence <220> <223> DNA site <400> 6 tagg 4 <210> 7 <211> 5 <212> DNA <213> Artificial Sequence <220> <223> DNA site <400> 7 atagg 5 <210> 8 <211> 4871 <212> DNA <213> Artificial Sequence <220> <223> B-mSA <400> 8 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac 60 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca 120 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta 180 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata 240 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag 300 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag 360 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg 420 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct 480 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc 540 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc 600 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca 660 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga 720 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa 780 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata 840 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc 900 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt 960 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat 1020 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta 1080 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt 1140 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca 1200 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta 1260 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcatggc ggaagcgggt 1320 atcaccggca cgtggtacaa ccagcatggt tctaccttca ccgttaccgc gggtgcggac 1380 ggtaacctga ccggtcagta cgaaaaccgt gcgcagggca ctggttgcca gaactctccg 1440 tacaccctga ccggtcgtta caacggtacc aaactggaat ggcgtgttga atggaacaac 1500 tctaccgaaa actgccactc tcgtaccgaa tggcgtggtc agtaccaggg tggtgcggaa 1560 gcgcgtatca acacccagtg gaacctgacc tacgaaggtg gttctggtcc ggcgaccgaa 1620 cagggtcagg acaccttcac caaagttaaa caccaccacc accaccacta agtcgacctg 1680 caggcatgca agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt 1740 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg 1800 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg 1860 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc 1920 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac 1980 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg 2040 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct 2100 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc 2160 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 2220 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 2280 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 2340 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 2400 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg 2460 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 2520 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 2580 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 2640 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 2700 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 2760 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 2820 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 2880 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 2940 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 3000 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 3060 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttacgcg 3120 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 3180 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 3240 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 3300 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg 3360 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 3420 ttgttccaaa cttgaacaac actcaaccct atctcgggct attcttttga tttataaggg 3480 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 3540 aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga tcctttttga 3600 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 3660 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 3720 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 3780 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta 3840 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 3900 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 3960 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 4020 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 4080 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 4140 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 4200 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 4260 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 4320 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 4380 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 4440 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca 4500 ccgcataggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg 4560 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 4620 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcaaggaga tggcgcccaa 4680 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc 4740 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc 4800 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt 4860 gacagcttat c 4871 <210> 9 <211> 4356 <212> DNA <213> Artificial Sequence <220> <223> B_R0.3-mSA <400> 9 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatacc cgttttttgg gctcacacag gaaaggctag catggcggaa gcgggtatca 3960 ccggcacgtg gtacaaccag catggttcta ccttcaccgt taccgcgggt gcggacggta 4020 acctgaccgg tcagtacgaa aaccgtgcgc agggcactgg ttgccagaac tctccgtaca 4080 ccctgaccgg tcgttacaac ggtaccaaac tggaatggcg tgttgaatgg aacaactcta 4140 ccgaaaactg ccactctcgt accgaatggc gtggtcagta ccagggtggt gcggaagcgc 4200 gtatcaacac ccagtggaac ctgacctacg aaggtggttc tggtccggcg accgaacagg 4260 gtcaggacac cttcaccaaa gttaaacacc accaccacca ccactaagtc gacctgcagg 4320 aagcttgttt aaacggtctc cagcttggct gttttg 4356 <210> 10 <211> 4358 <212> DNA <213> Artificial Sequence <220> <223> B_R0.6-mSA <400> 10 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatacc cgttttttgg gcattaaaga ggagaaagct agcatggcgg aagcgggtat 3960 caccggcacg tggtacaacc agcatggttc taccttcacc gttaccgcgg gtgcggacgg 4020 taacctgacc ggtcagtacg aaaaccgtgc gcagggcact ggttgccaga actctccgta 4080 caccctgacc ggtcgttaca acggtaccaa actggaatgg cgtgttgaat ggaacaactc 4140 taccgaaaac tgccactctc gtaccgaatg gcgtggtcag taccagggtg gtgcggaagc 4200 gcgtatcaac acccagtgga acctgaccta cgaaggtggt tctggtccgg cgaccgaaca 4260 gggtcaggac accttcacca aagttaaaca ccaccaccac caccactaag tcgacctgca 4320 ggaagcttgt ttaaacggtc tccagcttgg ctgttttg 4358 <210> 11 <211> 4355 <212> DNA <213> Artificial Sequence <220> <223> B_R1.0-mSA <400> 11 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatacc cgttttttgg gcaaagagga gaaagctagc atggcggaag cgggtatcac 3960 cggcacgtgg tacaaccagc atggttctac cttcaccgtt accgcgggtg cggacggtaa 4020 cctgaccggt cagtacgaaa accgtgcgca gggcactggt tgccagaact ctccgtacac 4080 cctgaccggt cgttacaacg gtaccaaact ggaatggcgt gttgaatgga acaactctac 4140 cgaaaactgc cactctcgta ccgaatggcg tggtcagtac cagggtggtg cggaagcgcg 4200 tatcaacacc cagtggaacc tgacctacga aggtggttct ggtccggcga ccgaacaggg 4260 tcaggacacc ttcaccaaag ttaaacacca ccaccaccac cactaagtcg acctgcagga 4320 agcttgttta aacggtctcc agcttggctg ttttg 4355 <210> 12 <211> 7057 <212> DNA <213> Artificial Sequence <220> <223> M_p-mSA <400> 12 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga 60 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg 120 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa 180 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac 240 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc 300 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg 360 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc 420 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca 480 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga 540 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc 600 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg 660 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag 720 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga 780 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa 840 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg 900 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc 960 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga 1020 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata 1080 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 1140 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag 1200 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg 1260 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg 1320 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt 1380 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga 1440 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga 1500 gcacttcacc aacaaggacc atagcatatg aaaataaaaa caggtgcacg catcctcgca 1560 ttatccgcat taacgacgat gatgttttcc gcctcggctc tcgccaaaat cgaagaaggt 1620 aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag 1680 aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa actggaagag 1740 aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac 1800 cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc 1860 caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct 1920 tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg 1980 ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc 2040 gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg 2100 ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac 2160 gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat 2220 gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc 2280 atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg 2340 gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt 2400 attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg 2460 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag 2520 tcttacgagg aagagttggc gaaagatcca cgtattgccg ccactatgga aaacgcccag 2580 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact 2640 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag 2700 actaattcga gctcgaacaa caacaacaat aacaataaca acaacctcgg gatcgaggga 2760 aggatttcag aattcgcgga agcgggtatc accggcacgt ggtacaacca gcatggttct 2820 accttcaccg ttaccgcggg tgcggacggt aacctgaccg gtcagtacga aaaccgtgcg 2880 cagggcactg gttgccagaa ctctccgtac accctgaccg gtcgttacaa cggtaccaaa 2940 ctggaatggc gtgttgaatg gaacaactct accgaaaact gccactctcg taccgaatgg 3000 cgtggtcagt accagggtgg tgcggaagcg cgtatcaaca cccagtggaa cctgacctac 3060 gaaggtggtt ctggtccggc gaccgaacag ggtcaggaca ccttcaccaa agttaaacac 3120 caccaccacc accactaaaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga 3180 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg 3240 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga 3300 atggcagctt ggctgttttg gcggatgaga taagattttc agcctgatac agattaaatc 3360 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc 3420 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc 3480 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag 3540 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc 3600 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc 3660 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg 3720 cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg 3780 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 3840 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 3900 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 3960 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttc 4020 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc 4080 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 4140 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 4200 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 4260 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 4320 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 4380 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 4440 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 4500 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 4560 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 4620 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 4680 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt accccggttg 4740 ataatcagaa aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaaacg 4800 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 4860 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 4920 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 4980 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt 5040 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 5100 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 5160 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 5220 ttaatgcgcc gctacagggc gcgtaaaagg atctaggtga agatcctttt tgataatctc 5280 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 5340 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 5400 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 5460 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag 5520 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 5580 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 5640 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 5700 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 5760 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 5820 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 5880 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 5940 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 6000 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 6060 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg 6120 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 6180 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc 6240 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc 6300 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg 6360 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta 6420 aagctcatca gcgtggtcgt gcagcgattc acagatgtct gcctgttcat ccgcgtccag 6480 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag 6540 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg 6600 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca 6660 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca 6720 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca 6780 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt 6840 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt 6900 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg 6960 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat 7020 gcgcacccgt ggccaggacc caacgctgcc cgaaatt 7057 <210> 13 <211> 6982 <212> DNA <213> Artificial Sequence <220> <223> M_c-mSA <400> 13 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga 60 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg 120 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa 180 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac 240 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc 300 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg 360 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc 420 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca 480 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga 540 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc 600 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg 660 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag 720 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga 780 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa 840 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg 900 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc 960 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga 1020 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata 1080 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 1140 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag 1200 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg 1260 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg 1320 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt 1380 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga 1440 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga 1500 gcacttcacc aacaaggacc atagcatatg aaaatcgaag aaggtaaact ggtaatctgg 1560 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat 1620 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt 1680 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac 1740 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat 1800 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt 1860 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa 1920 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac 1980 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag 2040 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg 2100 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac 2160 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg 2220 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc 2280 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt 2340 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2400 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2460 ttggcgaaag atccacgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg 2520 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2580 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2640 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcagaattc 2700 gcggaagcgg gtatcaccgg cacgtggtac aaccagcatg gttctacctt caccgttacc 2760 gcgggtgcgg acggtaacct gaccggtcag tacgaaaacc gtgcgcaggg cactggttgc 2820 cagaactctc cgtacaccct gaccggtcgt tacaacggta ccaaactgga atggcgtgtt 2880 gaatggaaca actctaccga aaactgccac tctcgtaccg aatggcgtgg tcagtaccag 2940 ggtggtgcgg aagcgcgtat caacacccag tggaacctga cctacgaagg tggttctggt 3000 ccggcgaccg aacagggtca ggacaccttc accaaagtta aacaccacca ccaccaccac 3060 taaaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac 3120 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc 3180 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc agcttggctg 3240 ttttggcgga tgagataaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg 3300 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc 3360 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt 3420 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt 3480 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt 3540 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca 3600 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc 3660 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 3720 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc 3780 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg 3840 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc 3900 aacagcggta agatccttga gagttttcgc cccgaagaac gtttcccaat gatgagcact 3960 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc 4020 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag 4080 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat 4140 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt 4200 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa 4260 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc 4320 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg 4380 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt 4440 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca 4500 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat 4560 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca 4620 gaccaagttt actcatatat actttagatt gatttacccc ggttgataat cagaaaagcc 4680 ccaaaaacag gaagattgta taagcaaata tttaaattgt aaacgttaat attttgttaa 4740 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca 4800 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga 4860 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc 4920 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc 4980 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc 5040 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg 5100 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac 5160 agggcgcgta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 5220 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 5280 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 5340 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 5400 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc 5460 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 5520 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 5580 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 5640 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 5700 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 5760 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 5820 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 5880 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 5940 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 6000 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg 6060 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca 6120 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga 6180 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg 6240 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca 6300 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg 6360 gtcgtgcagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc 6420 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg 6480 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat 6540 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga 6600 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca 6660 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca 6720 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact 6780 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca 6840 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca 6900 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca 6960 ggacccaacg ctgcccgaaa tt 6982 <210> 14 <211> 5606 <212> DNA <213> Artificial Sequence <220> <223> B_p-mSA <400> 14 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatacc cgttttttgg gctaacagga ggaagctagc gctagcatga aaataaaaac 3960 aggtgcacgc atcctcgcat tatccgcatt aacgacgatg atgttttccg cctcggctct 4020 cgccaaaatc gaagaaggta aactggtaat ctggattaac ggcgataaag gctataacgg 4080 tctcgctgaa gtcggtaaga aattcgagaa agataccgga attaaagtca ccgttgagca 4140 tccggataaa ctggaagaga aattcccaca ggttgcggca actggcgatg gccctgacat 4200 tatcttctgg gcacacgacc gctttggtgg ctacgctcaa tctggcctgt tggctgaaat 4260 caccccggac aaagcgttcc aggacaagct gtatccgttt acctgggatg ccgtacgtta 4320 caacggcaag ctgattgctt acccgatcgc tgttgaagcg ttatcgctga tttataacaa 4380 agatctgctg ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg ataaagaact 4440 gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact tcacctggcc 4500 gctgattgct gctgacgggg gttatgcgtt caagtatgaa aacggcaagt acgacattaa 4560 agacgtgggc gtggataacg ctggcgcgaa agcgggtctg accttcctgg ttgacctgat 4620 taaaaacaaa cacatgaatg cagacaccga ttactccatc gcagaagctg cctttaataa 4680 aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg tccaacatcg acaccagcaa 4740 agtgaattat ggtgtaacgg tactgccgac cttcaagggt caaccatcca aaccgttcgt 4800 tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac aaagagctgg caaaagagtt 4860 cctcgaaaac tatctgctga ctgatgaagg tctggaagcg gttaataaag acaaaccgct 4920 gggtgccgta gcgctgaagt cttacgagga agagttggcg aaagatccac gtattgccgc 4980 caccatggaa aacgcccaga aaggtgaaat catgccgaac atcccgcaga tgtccgcttt 5040 ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga ctgtcgatga 5100 agccctgaaa gacgcgcaga ctaattcgag ctcgaacaac aacaacaata acaataacaa 5160 caacctcggg atcgagggaa ggatttcaga attcgcggaa gcgggtatca ccggcacgtg 5220 gtacaaccag catggttcta ccttcaccgt taccgcgggt gcggacggta acctgaccgg 5280 tcagtacgaa aaccgtgcgc agggcactgg ttgccagaac tctccgtaca ccctgaccgg 5340 tcgttacaac ggtaccaaac tggaatggcg tgttgaatgg aacaactcta ccgaaaactg 5400 ccactctcgt accgaatggc gtggtcagta ccagggtggt gcggaagcgc gtatcaacac 5460 ccagtggaac ctgacctacg aaggtggttc tggtccggcg accgaacagg gtcaggacac 5520 cttcaccaaa gttaaacacc accaccacca ccactaagtc gacctgcagg aagcttgttt 5580 aaacggtctc cagcttggct gttttg 5606 <210> 15 <211> 5531 <212> DNA <213> Artificial Sequence <220> <223> B_c-mSA <400> 15 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatacc cgttttttgg gctaacagga ggaagctagc gctagcatga aaatcgaaga 3960 aggtaaactg gtaatctgga ttaacggcga taaaggctat aacggtctcg ctgaagtcgg 4020 taagaaattc gagaaagata ccggaattaa agtcaccgtt gagcatccgg ataaactgga 4080 agagaaattc ccacaggttg cggcaactgg cgatggccct gacattatct tctgggcaca 4140 cgaccgcttt ggtggctacg ctcaatctgg cctgttggct gaaatcaccc cggacaaagc 4200 gttccaggac aagctgtatc cgtttacctg ggatgccgta cgttacaacg gcaagctgat 4260 tgcttacccg atcgctgttg aagcgttatc gctgatttat aacaaagatc tgctgccgaa 4320 cccgccaaaa acctgggaag agatcccggc gctggataaa gaactgaaag cgaaaggtaa 4380 gagcgcgctg atgttcaacc tgcaagaacc gtacttcacc tggccgctga ttgctgctga 4440 cgggggttat gcgttcaagt atgaaaacgg caagtacgac attaaagacg tgggcgtgga 4500 taacgctggc gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa acaaacacat 4560 gaatgcagac accgattact ccatcgcaga agctgccttt aataaaggcg aaacagcgat 4620 gaccatcaac ggcccgtggg catggtccaa catcgacacc agcaaagtga attatggtgt 4680 aacggtactg ccgaccttca agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc 4740 aggtattaac gccgccagtc cgaacaaaga gctggcaaaa gagttcctcg aaaactatct 4800 gctgactgat gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct 4860 gaagtcttac gaggaagagt tggcgaaaga tccacgtatt gccgccacca tggaaaacgc 4920 ccagaaaggt gaaatcatgc cgaacatccc gcagatgtcc gctttctggt atgccgtgcg 4980 tactgcggtg atcaacgccg ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc 5040 gcagactaat tcgagctcga acaacaacaa caataacaat aacaacaacc tcgggatcga 5100 gggaaggatt tcagaattcg cggaagcggg tatcaccggc acgtggtaca accagcatgg 5160 ttctaccttc accgttaccg cgggtgcgga cggtaacctg accggtcagt acgaaaaccg 5220 tgcgcagggc actggttgcc agaactctcc gtacaccctg accggtcgtt acaacggtac 5280 caaactggaa tggcgtgttg aatggaacaa ctctaccgaa aactgccact ctcgtaccga 5340 atggcgtggt cagtaccagg gtggtgcgga agcgcgtatc aacacccagt ggaacctgac 5400 ctacgaaggt ggttctggtc cggcgaccga acagggtcag gacaccttca ccaaagttaa 5460 acaccaccac caccaccact aagtcgacct gcaggaagct tgtttaaacg gtctccagct 5520 tggctgtttt g 5531 <210> 16 <211> 5600 <212> DNA <213> Artificial Sequence <220> <223> B_R1.0-p-mSA <400> 16 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatacc cgttttttgg gcaaagagga gaaagctagc atgaaaataa aaacaggtgc 3960 acgcatcctc gcattatccg cattaacgac gatgatgttt tccgcctcgg ctctcgccaa 4020 aatcgaagaa ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc 4080 tgaagtcggt aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga 4140 taaactggaa gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt 4200 ctgggcacac gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc 4260 ggacaaagcg ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg 4320 caagctgatt gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct 4380 gctgccgaac ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc 4440 gaaaggtaag agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat 4500 tgctgctgac gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt 4560 gggcgtggat aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa 4620 caaacacatg aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga 4680 aacagcgatg accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa 4740 ttatggtgta acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt 4800 gctgagcgca ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga 4860 aaactatctg ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc 4920 cgtagcgctg aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccaccat 4980 ggaaaacgcc cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta 5040 tgccgtgcgt actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct 5100 gaaagacgcg cagactaatt cgagctcgaa caacaacaac aataacaata acaacaacct 5160 cgggatcgag ggaaggattt cagaattcgc ggaagcgggt atcaccggca cgtggtacaa 5220 ccagcatggt tctaccttca ccgttaccgc gggtgcggac ggtaacctga ccggtcagta 5280 cgaaaaccgt gcgcagggca ctggttgcca gaactctccg tacaccctga ccggtcgtta 5340 caacggtacc aaactggaat ggcgtgttga atggaacaac tctaccgaaa actgccactc 5400 tcgtaccgaa tggcgtggtc agtaccaggg tggtgcggaa gcgcgtatca acacccagtg 5460 gaacctgacc tacgaaggtg gttctggtcc ggcgaccgaa cagggtcagg acaccttcac 5520 caaagttaaa caccaccacc accaccacta agtcgacctg caggaagctt gtttaaacgg 5580 tctccagctt ggctgttttg 5600 <210> 17 <211> 5525 <212> DNA <213> Artificial Sequence <220> <223> B_R1.0-c-mSA <400> 17 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatacc cgttttttgg gcaaagagga gaaagctagc atgaaaatcg aagaaggtaa 3960 actggtaatc tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa 4020 attcgagaaa gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa 4080 attcccacag gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg 4140 ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca 4200 ggacaagctg tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta 4260 cccgatcgct gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc 4320 aaaaacctgg gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc 4380 gctgatgttc aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg 4440 ttatgcgttc aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc 4500 tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc 4560 agacaccgat tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat 4620 caacggcccg tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt 4680 actgccgacc ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat 4740 taacgccgcc agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac 4800 tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc 4860 ttacgaggaa gagttggcga aagatccacg tattgccgcc accatggaaa acgcccagaa 4920 aggtgaaatc atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc 4980 ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac 5040 taattcgagc tcgaacaaca acaacaataa caataacaac aacctcggga tcgagggaag 5100 gatttcagaa ttcgcggaag cgggtatcac cggcacgtgg tacaaccagc atggttctac 5160 cttcaccgtt accgcgggtg cggacggtaa cctgaccggt cagtacgaaa accgtgcgca 5220 gggcactggt tgccagaact ctccgtacac cctgaccggt cgttacaacg gtaccaaact 5280 ggaatggcgt gttgaatgga acaactctac cgaaaactgc cactctcgta ccgaatggcg 5340 tggtcagtac cagggtggtg cggaagcgcg tatcaacacc cagtggaacc tgacctacga 5400 aggtggttct ggtccggcga ccgaacaggg tcaggacacc ttcaccaaag ttaaacacca 5460 ccaccaccac cactaagtcg acctgcagga agcttgttta aacggtctcc agcttggctg 5520 ttttg 5525 <210> 18 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> B_R01-p-mSA <400> 18 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattag cactcgttga catacggacg tcacatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 19 <211> 5582 <212> DNA <213> Artificial Sequence <220> <223> B_R02-p-mSA <400> 19 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatact actgaggcta ctatgaaaat aaaaacaggt gcacgcatcc tcgcattatc 3960 cgcattaacg acgatgatgt tttccgcctc ggctctcgcc aaaatcgaag aaggtaaact 4020 ggtaatctgg attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt 4080 cgagaaagat accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt 4140 cccacaggtt gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt 4200 tggtggctac gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga 4260 caagctgtat ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc 4320 gatcgctgtt gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa 4380 aacctgggaa gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct 4440 gatgttcaac ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta 4500 tgcgttcaag tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg 4560 cgcgaaagcg ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga 4620 caccgattac tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa 4680 cggcccgtgg gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact 4740 gccgaccttc aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa 4800 cgccgccagt ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga 4860 tgaaggtctg gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta 4920 cgaggaagag ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg 4980 tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt 5040 gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa 5100 ttcgagctcg aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat 5160 ttcagaattc gcggaagcgg gtatcaccgg cacgtggtac aaccagcatg gttctacctt 5220 caccgttacc gcgggtgcgg acggtaacct gaccggtcag tacgaaaacc gtgcgcaggg 5280 cactggttgc cagaactctc cgtacaccct gaccggtcgt tacaacggta ccaaactgga 5340 atggcgtgtt gaatggaaca actctaccga aaactgccac tctcgtaccg aatggcgtgg 5400 tcagtaccag ggtggtgcgg aagcgcgtat caacacccag tggaacctga cctacgaagg 5460 tggttctggt ccggcgaccg aacagggtca ggacaccttc accaaagtta aacaccacca 5520 ccaccaccac taagtcgacc tgcaggaagc ttgtttaaac ggtctccagc ttggctgttt 5580 tg 5582 <210> 20 <211> 5597 <212> DNA <213> Artificial Sequence <220> <223> B_R1-p-mSA <400> 20 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattgg aacagctcac gcaaaaatag gtttcttatg aaaataaaaa caggtgcacg 3960 catcctcgca ttatccgcat taacgacgat gatgttttcc gcctcggctc tcgccaaaat 4020 cgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga 4080 agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa 4140 actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg 4200 ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga 4260 caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa 4320 gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct 4380 gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa 4440 aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc 4500 tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg 4560 cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa 4620 acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac 4680 agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta 4740 tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct 4800 gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa 4860 ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt 4920 agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg ccaccatgga 4980 aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc 5040 cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa 5100 agacgcgcag actaattcga gctcgaacaa caacaacaat aacaataaca acaacctcgg 5160 gatcgaggga aggatttcag aattcgcgga agcgggtatc accggcacgt ggtacaacca 5220 gcatggttct accttcaccg ttaccgcggg tgcggacggt aacctgaccg gtcagtacga 5280 aaaccgtgcg cagggcactg gttgccagaa ctctccgtac accctgaccg gtcgttacaa 5340 cggtaccaaa ctggaatggc gtgttgaatg gaacaactct accgaaaact gccactctcg 5400 taccgaatgg cgtggtcagt accagggtgg tgcggaagcg cgtatcaaca cccagtggaa 5460 cctgacctac gaaggtggtt ctggtccggc gaccgaacag ggtcaggaca ccttcaccaa 5520 agttaaacac caccaccacc accactaagt cgacctgcag gaagcttgtt taaacggtct 5580 ccagcttggc tgttttg 5597 <210> 21 <211> 5601 <212> DNA <213> Artificial Sequence <220> <223> B_R11-p-mSA <400> 21 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatcgc tttttatcgc aactctctac tgtttctcca tatgaaaata aaaacaggtg 3960 cacgcatcct cgcattatcc gcattaacga cgatgatgtt ttccgcctcg gctctcgcca 4020 aaatcgaaga aggtaaactg gtaatctgga ttaacggcga taaaggctat aacggtctcg 4080 ctgaagtcgg taagaaattc gagaaagata ccggaattaa agtcaccgtt gagcatccgg 4140 ataaactgga agagaaattc ccacaggttg cggcaactgg cgatggccct gacattatct 4200 tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg cctgttggct gaaatcaccc 4260 cggacaaagc gttccaggac aagctgtatc cgtttacctg ggatgccgta cgttacaacg 4320 gcaagctgat tgcttacccg atcgctgttg aagcgttatc gctgatttat aacaaagatc 4380 tgctgccgaa cccgccaaaa acctgggaag agatcccggc gctggataaa gaactgaaag 4440 cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc gtacttcacc tggccgctga 4500 ttgctgctga cgggggttat gcgttcaagt atgaaaacgg caagtacgac attaaagacg 4560 tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa 4620 acaaacacat gaatgcagac accgattact ccatcgcaga agctgccttt aataaaggcg 4680 aaacagcgat gaccatcaac ggcccgtggg catggtccaa catcgacacc agcaaagtga 4740 attatggtgt aacggtactg ccgaccttca agggtcaacc atccaaaccg ttcgttggcg 4800 tgctgagcgc aggtattaac gccgccagtc cgaacaaaga gctggcaaaa gagttcctcg 4860 aaaactatct gctgactgat gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg 4920 ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga tccacgtatt gccgccacca 4980 tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc gcagatgtcc gctttctggt 5040 atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg tcagactgtc gatgaagccc 5100 tgaaagacgc gcagactaat tcgagctcga acaacaacaa caataacaat aacaacaacc 5160 tcgggatcga gggaaggatt tcagaattcg cggaagcggg tatcaccggc acgtggtaca 5220 accagcatgg ttctaccttc accgttaccg cgggtgcgga cggtaacctg accggtcagt 5280 acgaaaaccg tgcgcagggc actggttgcc agaactctcc gtacaccctg accggtcgtt 5340 acaacggtac caaactggaa tggcgtgttg aatggaacaa ctctaccgaa aactgccact 5400 ctcgtaccga atggcgtggt cagtaccagg gtggtgcgga agcgcgtatc aacacccagt 5460 ggaacctgac ctacgaaggt ggttctggtc cggcgaccga acagggtcag gacaccttca 5520 ccaaagttaa acaccaccac caccaccact aagtcgacct gcaggaagct tgtttaaacg 5580 gtctccagct tggctgtttt g 5601 <210> 22 <211> 5592 <212> DNA <213> Artificial Sequence <220> <223> B_R12-p-mSA <400> 22 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct gagaaagaca cgatcttact agatgaaaat aaaaacaggt gcacgcatcc 3960 tcgcattatc cgcattaacg acgatgatgt tttccgcctc ggctctcgcc aaaatcgaag 4020 aaggtaaact ggtaatctgg attaacggcg ataaaggcta taacggtctc gctgaagtcg 4080 gtaagaaatt cgagaaagat accggaatta aagtcaccgt tgagcatccg gataaactgg 4140 aagagaaatt cccacaggtt gcggcaactg gcgatggccc tgacattatc ttctgggcac 4200 acgaccgctt tggtggctac gctcaatctg gcctgttggc tgaaatcacc ccggacaaag 4260 cgttccagga caagctgtat ccgtttacct gggatgccgt acgttacaac ggcaagctga 4320 ttgcttaccc gatcgctgtt gaagcgttat cgctgattta taacaaagat ctgctgccga 4380 acccgccaaa aacctgggaa gagatcccgg cgctggataa agaactgaaa gcgaaaggta 4440 agagcgcgct gatgttcaac ctgcaagaac cgtacttcac ctggccgctg attgctgctg 4500 acgggggtta tgcgttcaag tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg 4560 ataacgctgg cgcgaaagcg ggtctgacct tcctggttga cctgattaaa aacaaacaca 4620 tgaatgcaga caccgattac tccatcgcag aagctgcctt taataaaggc gaaacagcga 4680 tgaccatcaa cggcccgtgg gcatggtcca acatcgacac cagcaaagtg aattatggtg 4740 taacggtact gccgaccttc aagggtcaac catccaaacc gttcgttggc gtgctgagcg 4800 caggtattaa cgccgccagt ccgaacaaag agctggcaaa agagttcctc gaaaactatc 4860 tgctgactga tgaaggtctg gaagcggtta ataaagacaa accgctgggt gccgtagcgc 4920 tgaagtctta cgaggaagag ttggcgaaag atccacgtat tgccgccacc atggaaaacg 4980 cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc 5040 gtactgcggt gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg 5100 cgcagactaa ttcgagctcg aacaacaaca acaataacaa taacaacaac ctcgggatcg 5160 agggaaggat ttcagaattc gcggaagcgg gtatcaccgg cacgtggtac aaccagcatg 5220 gttctacctt caccgttacc gcgggtgcgg acggtaacct gaccggtcag tacgaaaacc 5280 gtgcgcaggg cactggttgc cagaactctc cgtacaccct gaccggtcgt tacaacggta 5340 ccaaactgga atggcgtgtt gaatggaaca actctaccga aaactgccac tctcgtaccg 5400 aatggcgtgg tcagtaccag ggtggtgcgg aagcgcgtat caacacccag tggaacctga 5460 cctacgaagg tggttctggt ccggcgaccg aacagggtca ggacaccttc accaaagtta 5520 aacaccacca ccaccaccac taagtcgacc tgcaggaagc ttgtttaaac ggtctccagc 5580 ttggctgttt tg 5592 <210> 23 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> B_R13-p-mSA <400> 23 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagag cggatcctac ctagatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 24 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> B_R2-p-mSA <400> 24 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagat aggagaatac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 25 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> B_R21-p-mSA <400> 25 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagag gcgacggtac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 26 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> BBa_B0032 <400> 26 tcacacagga aag 13 <210> 27 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> BBa_B0030 <400> 27 attaaagagg agaaa 15 <210> 28 <211> 12 <212> DNA <213> Artificial Sequence <220> <223> BBa_B0034 <400> 28 aaagaggaga aa 12 <210> 29 <211> 39 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 29 acccgttttt tgggctaaca ggaggaagct agcgctagc 39 <210> 30 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 30 tagcactcgt tgacatacgg acgtcac 27 <210> 31 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 31 actactgagg ctact 15 <210> 32 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 32 tggaacagct cacgcaaaaa taggtttctt 30 <210> 33 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 33 cgctttttat cgcaactctc tactgtttct ccat 34 <210> 34 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 34 tctgagaaag acacgatctt actag 25 <210> 35 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 35 tctagagaaa gagcggatcc tacctag 27 <210> 36 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 36 tctagagaaa gataggagaa tactag 26 <210> 37 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 37 tctagagaaa gaggcgacgg tactag 26 <210> 38 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-1-mSA <400> 38 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagag gcgagtgtac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 39 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-5-mSA <400> 39 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagat aggaggttac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 40 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-7-mSA <400> 40 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagag gggacactac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 41 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-10-mSA <400> 41 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagag cggaaactac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 42 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-11-mSA <400> 42 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatttc tagagaaaga tttgaatata ctagatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 43 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-12-mSA <400> 43 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagaa cggacattac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 44 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-13-mSA <400> 44 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagac atgactatac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 45 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-14-mSA <400> 45 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagaa ctgaagatac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 46 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-16-mSA <400> 46 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagag gcgatcctac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 47 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-17-mSA <400> 47 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagaa gagagcctac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 48 <211> 5593 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-1-18-mSA <400> 48 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattct agagaaagaa gagagcctac tagatgaaaa taaaaacagg tgcacgcatc 3960 ctcgcattat ccgcattaac gacgatgatg ttttccgcct cggctctcgc caaaatcgaa 4020 gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc 4080 ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg 4140 gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca 4200 cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa 4260 gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg 4320 attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg 4380 aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt 4440 aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct 4500 gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg 4560 gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac 4620 atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg 4680 atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt 4740 gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc 4800 gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat 4860 ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg 4920 ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac catggaaaac 4980 gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg 5040 cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac 5100 gcgcagacta attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc 5160 gagggaagga tttcagaatt cgcggaagcg ggtatcaccg gcacgtggta caaccagcat 5220 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac 5280 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt 5340 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc 5400 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg 5460 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt 5520 aaacaccacc accaccacca ctaagtcgac ctgcaggaag cttgtttaaa cggtctccag 5580 cttggctgtt ttg 5593 <210> 49 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-2-mSA <400> 49 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa ccctaataca ttaggagatc ttctatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 50 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-3-mSA <400> 50 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa ccctaataca ttaggacata ttctatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 51 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-4-mSA <400> 51 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa ccctaataca ttaggacatc atctatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 52 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-5-mSA <400> 52 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa ccctaataca taaggagatc atatatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 53 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-6-mSA <400> 53 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa ccctaataca taaggacata atatatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 54 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-7-mSA <400> 54 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa ccctaataca taaggagatt atctatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 55 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-8-mSA <400> 55 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa ccctaataca ttaggagatt atatatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 56 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-14-mSA <400> 56 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa ccctaataca taaggacatc ttatatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 57 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-16-mSA <400> 57 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa cactaataca ttaggagatc ttctatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 58 <211> 5594 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-2-17-mSA <400> 58 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccatgaa cactaataca taaggacata atatatgaaa ataaaaacag gtgcacgcat 3960 cctcgcatta tccgcattaa cgacgatgat gttttccgcc tcggctctcg ccaaaatcga 4020 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt 4080 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact 4140 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc 4200 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa 4260 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct 4320 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc 4380 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg 4440 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc 4500 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt 4560 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca 4620 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc 4680 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg 4740 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag 4800 cgcaggtatt aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta 4860 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc 4920 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa 4980 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt 5040 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga 5100 cgcgcagact aattcgagct cgaacaacaa caacaataac aataacaaca acctcgggat 5160 cgagggaagg atttcagaat tcgcggaagc gggtatcacc ggcacgtggt acaaccagca 5220 tggttctacc ttcaccgtta ccgcgggtgc ggacggtaac ctgaccggtc agtacgaaaa 5280 ccgtgcgcag ggcactggtt gccagaactc tccgtacacc ctgaccggtc gttacaacgg 5340 taccaaactg gaatggcgtg ttgaatggaa caactctacc gaaaactgcc actctcgtac 5400 cgaatggcgt ggtcagtacc agggtggtgc ggaagcgcgt atcaacaccc agtggaacct 5460 gacctacgaa ggtggttctg gtccggcgac cgaacagggt caggacacct tcaccaaagt 5520 taaacaccac caccaccacc actaagtcga cctgcaggaa gcttgtttaa acggtctcca 5580 gcttggctgt tttg 5594 <210> 59 <211> 5599 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-3-4-mSA <400> 59 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattta agtagttaaa cagggtatat aggggaagaa tgaaaataaa aacaggtgca 3960 cgcatcctcg cattatccgc attaacgacg atgatgtttt ccgcctcggc tctcgccaaa 4020 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct 4080 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat 4140 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc 4200 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg 4260 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc 4320 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg 4380 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg 4440 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt 4500 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg 4560 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac 4620 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa 4680 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat 4740 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg 4800 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa 4860 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc 4920 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccaccatg 4980 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat 5040 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg 5100 aaagacgcgc agactaattc gagctcgaac aacaacaaca ataacaataa caacaacctc 5160 gggatcgagg gaaggatttc agaattcgcg gaagcgggta tcaccggcac gtggtacaac 5220 cagcatggtt ctaccttcac cgttaccgcg ggtgcggacg gtaacctgac cggtcagtac 5280 gaaaaccgtg cgcagggcac tggttgccag aactctccgt acaccctgac cggtcgttac 5340 aacggtacca aactggaatg gcgtgttgaa tggaacaact ctaccgaaaa ctgccactct 5400 cgtaccgaat ggcgtggtca gtaccagggt ggtgcggaag cgcgtatcaa cacccagtgg 5460 aacctgacct acgaaggtgg ttctggtccg gcgaccgaac agggtcagga caccttcacc 5520 aaagttaaac accaccacca ccaccactaa gtcgacctgc aggaagcttg tttaaacggt 5580 ctccagcttg gctgttttg 5599 <210> 60 <211> 5599 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-3-5-mSA <400> 60 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattta agtagttaaa cagggtatat aggacgagaa tgaaaataaa aacaggtgca 3960 cgcatcctcg cattatccgc attaacgacg atgatgtttt ccgcctcggc tctcgccaaa 4020 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct 4080 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat 4140 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc 4200 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg 4260 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc 4320 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg 4380 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg 4440 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt 4500 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg 4560 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac 4620 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa 4680 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat 4740 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg 4800 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa 4860 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc 4920 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccaccatg 4980 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat 5040 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg 5100 aaagacgcgc agactaattc gagctcgaac aacaacaaca ataacaataa caacaacctc 5160 gggatcgagg gaaggatttc agaattcgcg gaagcgggta tcaccggcac gtggtacaac 5220 cagcatggtt ctaccttcac cgttaccgcg ggtgcggacg gtaacctgac cggtcagtac 5280 gaaaaccgtg cgcagggcac tggttgccag aactctccgt acaccctgac cggtcgttac 5340 aacggtacca aactggaatg gcgtgttgaa tggaacaact ctaccgaaaa ctgccactct 5400 cgtaccgaat ggcgtggtca gtaccagggt ggtgcggaag cgcgtatcaa cacccagtgg 5460 aacctgacct acgaaggtgg ttctggtccg gcgaccgaac agggtcagga caccttcacc 5520 aaagttaaac accaccacca ccaccactaa gtcgacctgc aggaagcttg tttaaacggt 5580 ctccagcttg gctgttttg 5599 <210> 61 <211> 5599 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-3-11-mSA <400> 61 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattta agtagttaaa cagggtatat agggctataa tgaaaataaa aacaggtgca 3960 cgcatcctcg cattatccgc attaacgacg atgatgtttt ccgcctcggc tctcgccaaa 4020 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct 4080 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat 4140 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc 4200 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg 4260 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc 4320 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg 4380 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg 4440 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt 4500 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg 4560 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac 4620 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa 4680 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat 4740 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg 4800 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa 4860 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc 4920 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccaccatg 4980 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat 5040 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg 5100 aaagacgcgc agactaattc gagctcgaac aacaacaaca ataacaataa caacaacctc 5160 gggatcgagg gaaggatttc agaattcgcg gaagcgggta tcaccggcac gtggtacaac 5220 cagcatggtt ctaccttcac cgttaccgcg ggtgcggacg gtaacctgac cggtcagtac 5280 gaaaaccgtg cgcagggcac tggttgccag aactctccgt acaccctgac cggtcgttac 5340 aacggtacca aactggaatg gcgtgttgaa tggaacaact ctaccgaaaa ctgccactct 5400 cgtaccgaat ggcgtggtca gtaccagggt ggtgcggaag cgcgtatcaa cacccagtgg 5460 aacctgacct acgaaggtgg ttctggtccg gcgaccgaac agggtcagga caccttcacc 5520 aaagttaaac accaccacca ccaccactaa gtcgacctgc aggaagcttg tttaaacggt 5580 ctccagcttg gctgttttg 5599 <210> 62 <211> 5599 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-3-13-mSA <400> 62 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattta agtagttaaa cagggtatat aggaggataa tgaaaataaa aacaggtgca 3960 cgcatcctcg cattatccgc attaacgacg atgatgtttt ccgcctcggc tctcgccaaa 4020 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct 4080 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat 4140 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc 4200 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg 4260 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc 4320 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg 4380 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg 4440 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt 4500 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg 4560 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac 4620 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa 4680 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat 4740 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg 4800 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa 4860 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc 4920 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccaccatg 4980 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat 5040 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg 5100 aaagacgcgc agactaattc gagctcgaac aacaacaaca ataacaataa caacaacctc 5160 gggatcgagg gaaggatttc agaattcgcg gaagcgggta tcaccggcac gtggtacaac 5220 cagcatggtt ctaccttcac cgttaccgcg ggtgcggacg gtaacctgac cggtcagtac 5280 gaaaaccgtg cgcagggcac tggttgccag aactctccgt acaccctgac cggtcgttac 5340 aacggtacca aactggaatg gcgtgttgaa tggaacaact ctaccgaaaa ctgccactct 5400 cgtaccgaat ggcgtggtca gtaccagggt ggtgcggaag cgcgtatcaa cacccagtgg 5460 aacctgacct acgaaggtgg ttctggtccg gcgaccgaac agggtcagga caccttcacc 5520 aaagttaaac accaccacca ccaccactaa gtcgacctgc aggaagcttg tttaaacggt 5580 ctccagcttg gctgttttg 5599 <210> 63 <211> 5599 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-3-18-mSA <400> 63 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattta agtagttaaa cagggtatat agggcgataa tgaaaataaa aacaggtgca 3960 cgcatcctcg cattatccgc attaacgacg atgatgtttt ccgcctcggc tctcgccaaa 4020 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct 4080 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat 4140 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc 4200 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg 4260 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc 4320 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg 4380 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg 4440 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt 4500 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg 4560 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac 4620 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa 4680 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat 4740 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg 4800 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa 4860 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc 4920 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccaccatg 4980 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat 5040 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg 5100 aaagacgcgc agactaattc gagctcgaac aacaacaaca ataacaataa caacaacctc 5160 gggatcgagg gaaggatttc agaattcgcg gaagcgggta tcaccggcac gtggtacaac 5220 cagcatggtt ctaccttcac cgttaccgcg ggtgcggacg gtaacctgac cggtcagtac 5280 gaaaaccgtg cgcagggcac tggttgccag aactctccgt acaccctgac cggtcgttac 5340 aacggtacca aactggaatg gcgtgttgaa tggaacaact ctaccgaaaa ctgccactct 5400 cgtaccgaat ggcgtggtca gtaccagggt ggtgcggaag cgcgtatcaa cacccagtgg 5460 aacctgacct acgaaggtgg ttctggtccg gcgaccgaac agggtcagga caccttcacc 5520 aaagttaaac accaccacca ccaccactaa gtcgacctgc aggaagcttg tttaaacggt 5580 ctccagcttg gctgttttg 5599 <210> 64 <211> 5599 <212> DNA <213> Artificial Sequence <220> <223> pBAD_R-lib-3-20-mSA <400> 64 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga 60 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact 120 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga 180 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 240 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac 300 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat 360 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg 420 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 480 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 540 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 600 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 660 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 720 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 780 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 840 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 900 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 960 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 1020 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 1080 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 1140 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 1200 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 1260 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1320 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1380 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1440 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1500 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1560 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1620 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1680 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1740 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1800 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1860 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1920 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1980 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 2040 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 2100 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 2160 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 2220 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 2280 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2340 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 2400 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 2460 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 2520 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 2580 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc 2640 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt 2700 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa 2760 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag 2820 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc 2880 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct 2940 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg 3000 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt 3060 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac 3120 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg 3180 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag 3240 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga 3300 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga 3360 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga 3420 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc 3480 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc 3540 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt 3600 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt 3660 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc 3720 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat 3780 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca 3840 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt 3900 tctccattta agtaattaaa cagggtatat aggggaagaa tgaaaataaa aacaggtgca 3960 cgcatcctcg cattatccgc attaacgacg atgatgtttt ccgcctcggc tctcgccaaa 4020 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct 4080 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat 4140 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc 4200 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg 4260 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc 4320 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg 4380 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg 4440 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt 4500 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg 4560 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac 4620 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa 4680 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat 4740 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg 4800 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa 4860 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc 4920 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccaccatg 4980 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat 5040 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg 5100 aaagacgcgc agactaattc gagctcgaac aacaacaaca ataacaataa caacaacctc 5160 gggatcgagg gaaggatttc agaattcgcg gaagcgggta tcaccggcac gtggtacaac 5220 cagcatggtt ctaccttcac cgttaccgcg ggtgcggacg gtaacctgac cggtcagtac 5280 gaaaaccgtg cgcagggcac tggttgccag aactctccgt acaccctgac cggtcgttac 5340 aacggtacca aactggaatg gcgtgttgaa tggaacaact ctaccgaaaa ctgccactct 5400 cgtaccgaat ggcgtggtca gtaccagggt ggtgcggaag cgcgtatcaa cacccagtgg 5460 aacctgacct acgaaggtgg ttctggtccg gcgaccgaac agggtcagga caccttcacc 5520 aaagttaaac accaccacca ccaccactaa gtcgacctgc aggaagcttg tttaaacggt 5580 ctccagcttg gctgttttg 5599 <210> 65 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 65 tctagagaaa gaggcgagtg tactag 26 <210> 66 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 66 tctagagaaa gataggaggt tactag 26 <210> 67 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 67 tctagagaaa gaggggacac tactag 26 <210> 68 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 68 tctagagaaa gagcggaaac tactag 26 <210> 69 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 69 ttctagagaa agatttgaat atactag 27 <210> 70 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 70 tctagagaaa gaacggacat tactag 26 <210> 71 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 71 tctagagaaa gacatgacta tactag 26 <210> 72 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 72 tctagagaaa gaactgaaga tactag 26 <210> 73 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 73 tctagagaaa gaggcgatcc tactag 26 <210> 74 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 74 tctagagaaa gaagagagcc tactag 26 <210> 75 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 75 tctagagaaa gacttgaggc tactag 26 <210> 76 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 76 gaaccctaat acattaggag atcttct 27 <210> 77 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 77 gaaccctaat acattaggac atattct 27 <210> 78 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 78 gaaccctaat acattaggac atcatct 27 <210> 79 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 79 gaaccctaat acataaggag atcatat 27 <210> 80 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 80 gaaccctaat acataaggac ataatat 27 <210> 81 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 81 gaaccctaat acataaggag attatct 27 <210> 82 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 82 gaaccctaat acattaggag attatat 27 <210> 83 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 83 gaaccctaat acataaggac atcttat 27 <210> 84 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 84 gaacactaat acattaggag atcttct 27 <210> 85 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 85 gaacactaat acataaggac ataatat 27 <210> 86 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 86 ttaagtagtt aaacagggta tataggggaa ga 32 <210> 87 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 87 ttaagtagtt aaacagggta tataggacga ga 32 <210> 88 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 88 ttaagtagtt aaacagggta tatagggcta ta 32 <210> 89 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 89 ttaagtagtt aaacagggta tataggagga ta 32 <210> 90 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 90 ttaagtagtt aaacagggta tatagggcga ta 32 <210> 91 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> regulatory gene <400> 91 ttaagtaatt aaacagggta tataggggaa ga 32

Claims (24)

  1. 모노머형 스트렙타비딘(monomeric streptavidin; mSA)을 코딩하는 유전자가 도입되어 형질 전환된 숙주 세포를 포함하는, 생체 내 세포 추적용 조성물.
  2. 제 1항에 있어서,
    상기 숙주 세포는 말토오즈 결합 단백질(maltose-binding protein; MBP)을 코딩하는 유전자가 더 도입된 것인, 조성물.
  3. 제 1항에 있어서,
    상기 숙주 세포는 상기 모노머형 스트렙타비딘을 코딩하는 유전자의 발현을 조절하는 조절 유전자가 더 도입된 것인, 조성물.
  4. 제 3항에 있어서,
    상기 조절 유전자는 리보솜 결합 부위(Ribosome binding site; RBS), 5'-비 번역 영역(5'-Untransrated Region; 5'-UTR) 및 전사 인자 결합 부위(transcription factor binding site) 및 유도성 프로모터(Inducible promoter)로 이루어진 군으로부터 선택되는 적어도 하나인 것인, 조성물.
  5. 제 3항에 있어서,
    상기 조절 유전자를 포함하는 재조합 벡터가 상기 숙주 세포에 형질 전환되는 경우, 상기 숙주 세포의 주변 세포질로 모노머형 스트렙타비딘이 발현되도록 하는 것인, 조성물.
  6. 제 3항에 있어서,
    상기 조절 유전자는 총 깁스 자유 에너지 변화량(ΔGtotal)이 0 이하인 것인, 조성물.
  7. 제 3항에 있어서,
    상기 조절 유전자는 번역 개시 속도(Translation Initiation Rate; TIR)가 특정 범위를 갖도록 조절된 것인, 조성물.
  8. 제 3항에 있어서,
    상기 조절 유전자의 서열 길이는 15 내지 39bp인 것인, 조성물.
  9. 제 3항에 있어서,
    상기 조절 유전자는 서열번호 5 내지 7 중 어느 하나로 표시되는 유전자 서열을 포함하는 것인, 조성물.
  10. 제 9항에 있어서,
    상기 조절 유전자에서, 상기 서열번호 5 내지 7 중 어느 하나로 표시되는 유전자 서열의 3' 말단에서 상기 모노머형 스트렙타비딘을 코딩하는 유전자의 개시코돈 까지의 간격이 6 내지 13bp인, 조성물.
  11. 제 1항에 있어서,
    상기 숙주 세포는 박테리아, 효모, 균류 세포, 식물 세포, 곤충 세포 및 동물 세포로 이루어진 군으로부터 선택되는 어느 하나 이상의 세포인 것인, 조성물.
  12. 제 11항에 있어서,
    상기 박테리아는 락토코쿠스(Lactococcus), 류코노스톡(Leuconostoc), 페디오코커스(Pediococcus), 엔트로코커스(Enterococcus), 스트렙토코커스(Streptococcus), 베일로넬라(Veilonella), 에스체리시아(Escherichia), 유박테리움(Eubacterium), 슈도모나스(Pseudomonas), 살모넬라(Salmonella), 시겔라(Shigella), 헬리코박터(Helicobacter), 캄필로박터(Campylobacter), 어시니아(Yersinia), 리스테리아(Listeria), 스트렙토미세스(Streptomyces), 펩토코커스(Peptococcus), 펩토스트렙토코커스(Peptostreptococcus), 프로투스(Proteus), 루미노코커스(Ruminococcus), 엔테로박터(Enterobacter), 시트로박터(Citrobacter), 세라티아(Serratia), 해모필러브(Haemophilus), 스타필로코커스(Staphylococcus), 마이코박테리움(Mycobacterium), 클로스트리디움(Clostridium), 바실러스(Bacillus), 마이크로코커스(Micrococcus), 비브리오(Vibrio), 박테로이데스(Bacteroides), 멜리소코코스(Melissococcus), 코쿠리아(Kocuria), 아에로코코스(Aerococcus), 오에노코코스(Oenococcus), 락토바실러스(Lactobacillus), 스포로락토바실러스(Sporolactobacilus), 아커만시아(Akkermansia), 비피도박테리움(Bifidobacterium), 부티리치코쿠스(Butyricicoccus), 부티리시모나스(Butyricimonas), 부티비브리오(Butyrivibrio), 슈도부티비브리오(Pseudobutyrivibrio), 웨이셀라(Weissella), 푸소박테리움(Fusobacterium), 카르노박테리움(Carnobacterium), 프로피오니박테리움(Propionibacterium), 메가스파에라(Megasphaera), 알리스팁스(alistipes), 알로바쿨룸(allobaculum), 바네시엘라(barnesiella), 블라우티아(blautia), 도레아(dorea), 헤스펠리아(hespellia), 홀데마니아(holdemania), 라우소니아(lawsonia), 오실리박터(oscillibacter), 파라박테로이드(parabacteroides), 파스콜락토박테리움(phascolarctobacterium), 프리보텔라(prevotella), 세디멘티박터(sedimentibacter), 엑시구오박데리움(Exiguobacterium), 아시네토박터(Acinetobacter), 카프노시토파가(Capnocytophaga), 네이세리아(Neisseria), 스핑고모나스(Sphingomonas), 아그레가티박터(Aggregatibacter), 렙토트리키아(Leptotrichia), 그라눌리카텔라(Granulicatella), 크리세오박테리움(Chryseobacterium), 포르피로모나스(Porphyromonas), 브라키박테리움(Brachybacterium), 엔하이드로박터(Enhydrobacter), 파라코커스(Paracoccus), 코리네박테리움(Corynebacterium), 로티아(Rothia), 엑티노마이세스(Actinomyces), 디알리스터(Dialister), 페칼리박테리움(Faecalibacterium), 할로모나스(Halomonas), 수테렐라(Sutterella), 베일로넬라(Veillonella), 로도코커스(Rhodococcus), 아토포비움(Atopobium), 크로모할로박터(Chromohalobacter), 쿠프리아비두스(Cupriavidus), 메타노브레비박터(Methanobrevibacter), 오도리박터(Odoribacter), 피라미도박터(Pyramidobacter), 빌로필라(Bilophila), 데설포비브리오(Desulfovibrio), 아시다미노코커스(Acidaminococcus), 아크로모박터(Achromobacter), 아그로박테리움(Agrobacterium), 로시텔레스(Roseateles), 코프로코커스(Coprococcus), 터리시박터(Turicibacter), 로즈뷰리아(Roseburia), 라크노스피라(Lachnospira), 오스실로스피라(Oscillospira), SMB53, 카테니박테리움(Catenibacterium), 파라프레보텔라(Paraprevotella), 아들러크레우치아(Adlercreutzia), 슬라키아(Slackia) 및 데르모아나데로박테리움(Thermoanaerobacterium)에서 선택된 어느 하나 이상인 것인, 조성물.
  13. 제 11항에 있어서,
    상기 박테리아는 aroA, aroC, aroD, aroE, Rpur, htrA, ompR, ompF, ompC, galE, cya, crp, cyp, phoP, phoQ, rfaY, dksA, hupA, sipC, clpB, clpP, clpX, pab, nadA, pncB, pmi, rpsL, hemA, rfc, poxA, galU, cdt, pur, ssa, guaA, guaB, fliD, flgK, flgL, relA, spoA 및 spoT로 이루어진 군으로부터 선택되는 적어도 하나를 코딩하는 유전자가 변형된 것인, 조성물.
  14. 제 11항에 있어서,
    상기 박테리아는 구아노신-5-이인산-3-이인산(ppGpp)의 합성능이 결여된 것인, 조성물.
  15. 제 1항에 있어서,
    상기 조성물은 비오틴화된 화합물을 더 포함하는 것인, 조성물.
  16. 제 15항에 있어서,
    상기 화합물은 조영제인 것인, 조성물.
  17. 제 16항에 있어서,
    상기 조영제는 방사성 핵종, 형광 표지, 효소 표지, 화학 발광 마커, 금 제제 및 자성 제제로 이루어진 군으로부터 선택되는 적어도 하나인 것인, 조성물.
  18. 제 17항에 있어서,
    상기 방사성 핵종을 검출할 수 있는 수단은, 양전자방출단층촬영(PET) 또는 단일광자방출전산화 단층촬영(SPECT)인 것인, 조성물.
  19. 제 1항에 있어서,
    상기 조성물은 아라비노스를 더 포함하는 것인, 조성물.
  20. 모노머형 스트렙타비딘을 발현하는 숙주 세포가 투여된 목적하는 개체에서, 상기 숙주 세포를 촬영 수단으로 확인하는 단계를 포함하는, 생체 내 세포의 분포를 확인하기 위한 정보 제공 방법.
  21. 제 20항에 있어서,
    상기 숙주 세포는 박테리아, 효모, 균류 세포, 식물 세포, 곤충 세포 및 동물 세포로 이루어진 군으로부터 선택되는 어느 하나 이상의 세포인 것인, 정보 제공 방법.
  22. 제 20항에 있어서,
    상기 목적하는 개체는 비오틴화된 화합물이 더 투여된 것인, 정보 제공 방법.
  23. 제 20항에 있어서,
    상기 목적하는 개체는 아라비노스가 더 투여된 것인, 정보 제공 방법.
  24. 제 20항에 있어서,
    상기 확인하는 단계는 복수 회에 걸쳐 수행되는, 정보 제공 방법.
KR1020210146377A 2021-01-18 2021-10-29 예비표적화 단백질을 발현하는 균주를 포함하는 생체 내 미생물 추적 플랫폼 KR20220104626A (ko)

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