KR20120034595A

KR20120034595A - 환자맞춤형 줄기세포치료를 위한 세포투과성 역분화 전사인자를 이용한 유도만능줄기세포 확립

Info

Publication number: KR20120034595A
Application number: KR1020117023922A
Authority: KR
Inventors: 조대웅; 임정희; 김정은; 정수영; 정인희
Original assignee: 주식회사 프로셀제약; 조대웅
Priority date: 2009-03-12
Filing date: 2010-03-12
Publication date: 2012-04-12
Also published as: WO2010104357A2; WO2010104357A3; WO2010104357A4; CN102439045A; EP2407488A2; EP2407488A4; US20120095188A1

Abstract

본발명은 역분화 유도전사인자 (Reprogramming Transcription Factor)에 거대분자 전송 도메인 (macromolecule transduction domain, MTD)이 융합되어 세포투과성을 갖는 역분화 전사인자 재조합 단백질, 상기 역분화 전사인자 재조합 단백질을 코딩하는 폴리뉴클레오티드, 상기 세포투과성 역분화 전사인자 재조합 단백질의 발현벡터에 관한 것이다. 상기 세포투과성 역분화 전사인자 재조합 단백질이 체세포에 처리되면 체세포의 줄기세포 특이적 유전자의 리프로그래밍(Reprogramming)을 유도하여 외형적(morphology), 유전적(genetic)으로 배아줄기세포와 유사한 특징을 가지고 있는 유도만능줄기세포 (iPS cell)를 확립하는데 유용하게 사용될 수 있다.

Description

환자맞춤형 줄기세포치료를 위한 세포투과성 역분화 전사인자를 이용한 유도만능줄기세포 확립{ESTABLISHMENT OF INDUCED PLURIPOTENT STEM CELL USING CELL-PERMEABLE REPROGRAMMING TRANSCRIPTION FACTOR FOR CUSTOMIZED STEM CELL THERAPY}

본 발명은 역분화 유도전사인자 (Reprogramming Transcription Factor)에 거대분자 전송 도메인 (macromolecule transduction domain, MTD)이 융합되어 세포투과성을 갖는 역분화 전사인자 재조합 단백질, 상기 세포투과성 역분화 전사인자 재조합 단백질을 코딩하는 폴리뉴클레오티드, 상기 세포투과성 역분화 전사인자 재조합 단백질의 발현벡터에 관한 것이다. 상기 세포투과성 역분화 전사인자 재조합 단백질은 체세포의 역분화능을 효율적으로 증가시켜 유도만능줄기세포 (iPS cell)를 확립하는데 유용하게 사용될 수 있다.

배아줄기세포란 Blastocyst의 내부세포괴 (inner cell mass)로부터 유래한 'Pluripotent Stem Cell (전능성세포)' 이다. 배아줄기세포는 인체를 구성하는 내배엽, 중배엽, 외배엽으로 분화할 수 있으며, 내배엽은 갑상선을 이루는 세포, 폐를 구성하는 세포, 췌장세포로 발달하며 중배엽은 심근세포, 근육세포, 신장세포, 혈액세포, 외배엽은 피부세포, 신경세포와 색소세포로 각 분화하며 생식 세포인 정자와 난자로 분화도 가능하다. 배아줄기세포는 다양한 세포로의 분화가 가능하여 난치병을 치료할 수 있는 도구로써 매우 중요한 역할을 할 것으로 기대된다.

배아줄기세포의 확립 방법은 냉동인간배아 세포의 사용, 또는 핵이 제거된 난자에 체세포의 핵을 치환하는 체세포 핵치환 (Somatic Cell Nuclear Transfer)의 방법이 있다. 그러나 이들 두 방법은 인간의 배아에서 유래되는 세포를 사용한다는 점과 인간의 난자가 실험에 다량 쓰인다는 점에서 윤리적인 문제점이 있고, 낮은 핵 이식 효율과 복제로 인한 치명적인 유전자 결함 가능성을 가지고 있다.

인간배아와 난자 없이 배아줄기세포를 확립하는 다른 하나의 방법은 역분화 전사인자로 체세포를 역분화 시키는 방법이다. 이는 배아줄기세포의 유지에 관여하는 24개의 유전자를 선정하고 바이러스 벡터 전송법을 이용해 그 중 역분화에 관여하는 4개의 유전자 (Oct4, Sox2, c-Myc, Klf4)로 역분화를 유도하는 연구가 알려져 있다 (Yamanaka, Cell 126: 633-678, 2006). 레트로바이러스 (retrovirus)에 의해 전달된 이들 4가지 역분화 전사인자는 쥐섬유아세포 내에서 발현되어 체세포를 역분화시켜 유도만능줄기세포를 유도하였다. 또한, 또 다른 연구에서는 이 4가지의 역분화 전사인자 중 Oct4, Sox2와 새로운 인자인 Lin28과 Nanog를 렌티바이러스로 전달하여 인간 체세포를 역분화 하는데 성공하였다 (Yu J, Science 318(5858): 1917-20,2007). 따라서 역분화에 사용된 전사인자 6종(Oct4, Sox2, c-Myc, Klf4, Nanog, Lin28) 중에 Oct4와 Sox2는 공통으로 사용되고, c-Myc, Klf4, Nanog, Lin28은 다양한 조합으로 유도만능줄기세포를 확립하며 역분화 과정에 있어서 중요한 역할을 한다. 또한, 상기 유전자는 배아줄기세포의 전능성 (pluripotency)과 자가증식 (self renewal), 미분화 (undifferentiation)유지에 필수적이며, 이들이 발현한 전사인자는 서로 활성화 시키고 분화를 유도하고 자가증식을 억제하는 유전자를 저해하는 역할을 한다.

Oct-4 전사인자는 옥타머 전사인자 (octamer transcription factor) 가운데 하나로 POU (Pit-Oct-Unc) 패밀리의 클래스 V POU 도메인 구성원이다. Oct4는 미분화 세포인 배아줄기세포 및 배아종양세포에서만 발현되고 줄기세포의 만능성 (pluripotency)을 유지하는데 중요한 역할을 한다. Oct-4 발현이 정상 수준인 경우 배아줄기세포의 자가 재생산과 만능성이 유지되지만, 정상 수준 이상으로 증가하게 되면 원시 내배엽 (primitive endoderm)과 원시 중배엽 (primitive mesoderm)으로 분화하고, 반대로 정상 수준 이하로 감소하게 되면 영양외배엽 (trophectoderm) 세포로 발달하므로, Oct-4가 배아줄기세포의 만능성을 결정하고 분화를 조절하는 중요한 인자이며, Oct-4는 Sox2 전사인자와 상호작용을 통해 Oct-4의 하위 유전자인 Fgf4, Utf-1, Nanog와 Sox2 유전자들의 발현을 증가시키는 상승효과를 보인다.

Sox 패밀리의 유전자들은 다능성 줄기세포 (multipotent stem cell)나 단능성 줄기세포 (unipotent stem cell)와도 관련이되어 있으며 그 중 Sox2 (SRY-type high mobility group box 2) 전사인자는 HMG box DNA 결합 모티프를 포함하는 Sox 패밀리 구성원 중 하나로, 20개의 Sox 패밀리 단백질 중 유일하게 배아줄기세포의 만능성 유지에 중요한 역할을 한다. Oct-4와 마찬가지로 생쥐의 배아줄기세포에서 Sox2의 발현을 저해하면 분화가 유도되며 여러 Sox2 하위 유전자의 프로모터 영역에서 발견되는 Sox2 결합 부위는 종종 Oct-4와 Nanog 결합 부위와 인접하게 존재한다. 그러므로 Sox2 전사인자와 Oct-4 전사인자 사이의 상호 작용이 역분화 줄기세포가 미분화 상태를 유지하고 배아줄기세포의 특성을 지니게 한다.

Helix-loop-helix/leucine zipper 전사인자인 c-Myc은 세포 성장, 분화, 증식, 세포예정사 및 암세포로의 형질 전환 등의 다양한 세포 내 기능을 수행하는 발암 유전자이며, 만능성을 유지하는 주요 기작인 LIF (leukaemia inhibitory factor)/STAT3와 Wnt 신호 전달 메커니즘의 하위 유전자이다. c-Myc 전사인자는 역분화 줄기세포 내에서 Klf4 유전자에 의해 p21 유전자가 활성화되어 세포의 증식을 저해하는 것을 막아주는 역할을 한다. 또한, c-Myc은 게놈 상에 존재하는 Myc 인식부위에 결합할 뿐 아니라 histone acetylase 컴플렉스를 끌어옴으로써 염색질 구조를 변화시켜 Oct-4와 Sox2가 표적 유전자에 잘 결합할 수 있도록 도와 주는 기능을 수행한다.

Klf 패밀리의 한 멤버인 Klf4는 Yamanaka 그룹과 Jaenisch 그룹에 의해 생쥐와 인간의 역분화 줄기세포를 제작하는데 사용되었다. 그러나 Thomson 그룹은 Klf4 없이도 인간 역분화 줄기세포를 제작하였으므로, Klf4가 인간 역분화 줄기세포 제작에 반드시 필요한 유전자는 아닌 것으로 추정된다. Klf4는 Kruppel-type zinc-finger 전사인자로써 성장 억제에 관여하여 세포 주기를 조절하는 역할을 한다. c-Myc과 유사하게 배아줄기세포에서 STAT3의 하위 유전자로 작용하며, 과발현 시 Oct-4 발현을 유지시켜 생쥐 배아줄기세포의 분화를 억제한다고 밝혀졌다. 또한, Klf4는 배아줄기세포에서 Oct-4 하위 유전자로 알려진 Lefty1의 발현 조절에 Sox2와 Klf4가 관여하고 있으며 p53 항암 유전자의 기능을 억제한다. 따라서 Klf4가 간접적인 기작으로 배아 특이적인 유전인자인 Nanog의 활성을 증가시키고, c-Myc에 의해 유도되는 apoptosis를 억제하여 역분화 줄기세포 유도에 역할을 할 것으로 예상된다. Klf4뿐만 아니라 Klf2도 인간 역분화 줄기세포 제작에 사용할 수 있고, 관련 유전자인 Klf1과 Klf5도 인간 역분화 줄기세포 제작에 사용할 수 있지만 Klf4나 Klf2를 사용할 때보다 효율이 낮다고 보고되었다.

Nanog는 LIF/STAT3의 메카니즘을 거치지 않고 독립적으로 배아 특이적인 유전인자로 Oct-4나 Sox2와 마찬가지로 배아줄기세포의 만능성을 유지하는데 필요한 유전자이며, 내배엽으로 분화시키는 GATA4와 GATA6의 전사인자를 억제시키는 능력을 가지고 있다.

마지막으로 LIN28은 mRNA 결합 단백질로 배아줄기세포와 배아종양세포에서 발현되며 분화와 증식에 관여하고 있다고 알려져 있다. 이 두 유전자가 역분화 줄기세포 제작에 반드시 필요한 것은 아니지만 Thomson 박사 연구팀은 Nanog와 LIN28을 이용해 역분화 줄기세포 확립에 성공하였다.

이러한 6가지 역분화 전사인자를 이용해 역분화 줄기세포를 확립하는 연구는 전사인자의 체내 전송이 가장 큰 핵심으로 현재 다양하게 연구되어지고 있다. 이중 바이러스 벡터를 이용한 역분화 전사인자의 전송방법이 일반적으로 이용된다. 그러나, 바이러스 벡터 전송법은 종양이 발생하는 부작용을 동반하는데, 6가지의 역분화 전사인자 (Oct4, Sox2, c-Myc, Klf4, Nanog, Lin28)를 코딩하는 레트로바이러스 (retrovirus), 렌티바이러스 (lentivirus)가 체세포의 유전자에 삽입되면 외부 유전자의 간섭으로 인해 체세포 유전자의 돌연변이 (mutagenesis)를 일으키면서, 발암유전자 (oncogene)인 c-Myc 유전자에 의해 역분화 후 태어난 카이메라(chimera)의 25％ 개체에서 이형종양(teratoma)이 발생됨이 보고 되었다.

이러한 바이러스 혹은 플라스미드의 역분화 전사인자 전송 방법은 돌연변이와 종양형성의 가능성으로 인해 세포치료에 적용하는데 한계가 있다. 따라서 단백질 전송법을 적용한 세포투과성 역분화 전사인자의 전송방법은 외부 유전자의 삽입이 없는 안전한 방법이다. 이에, 본 발명에서는 "거대분자 전달 도메인 (Macromolecule Transduction Domain, MTD)"을 융합시켜 세포투과성을 부여한 역분화 전사인자 재조합 단백질 (Cell Permeable CP-TFs)을 제조하여 역분화능을 가진 전사인자를 만들고자 하였다.

한편, 합성화합물 (synthetic compound) 또는 천연물의 작은 분자들 (small molecules)은 세포 안으로 전달될 수 있으나, 단백질, 펩티드, 핵산같은 거대분자들 (macromolecules)은 분자량 때문에 세포 내로 전송될 수 없다. 이러한 거대 분자들은 분자량 500 이상의 거대분자로 살아있는 세포의 원형질막 (plasma membrane), 즉 이중 지질막 구조 (lipid bilayer structure)를 통과할 수 없음이 널리 알려져 있다. 이를 극복하기 위한 방법으로 "거대분자 세포 내 전달기술 (macromolecule intracellular transduction technology: MITT)"이 개발되었는데 (한국특허출원: 제10-2009-7017564호, 미국특허출원: 12/524935, 캐나다특허출원: 제2,676,797호, 중국특허출원: 제200880003468.9호, 호주특허출원: 제2008211854호, 인도특허출원: 5079/CHENP/2009, EU특허출원: 제08712219.8호, 일본특허출원: 제2009-547177호, (USPP: 60/887,060에 기초한 우선권 주장)), 이는 치료용 거대분자들 (therapeutic macromolecule)의 생체 내 전달을 가능케 함으로써 기존의 의약품 개발 기술로는 불가능했던 펩티드, 단백질, 유전자 자체 등을 이용한 바이오 신약물의 개발을 가능하게 한다. 이 기술에 따르면, 거대분자들은 "소수성 거대분자 전달 도메인 (hydrophobic macromolecule transduction domain)"과 다른 여러 가지 세포 내 운반체들과 융합되어 재조합 단백질 형태로 합성 또는 발현, 정제되고, 생체 내 투여되어 세포 내로 전달된 후, 특정 세포 내 위치로 정확하고 빠르게 운반되어 필요한 여러 가지 역할을 효과적으로 발휘할 수 있다. 이처럼 거대분자 전달 도메인 (MTD)은 펩티드, 단백질, DNA, RNA, 합성화합물 등과 융합되어, 세포 내로 들어갈 수 없는 많은 불투과성 물질들의 전송을 가능케 한다.

이에 본 발명자들은 역분화 전사인자 6종 (Nanog, Oct4, Sox2, Klf4, c-Myc 및 Lin28)에 거대분자 전달 도메인 (MTD)을 융합시켜 세포투과성을 부여한 역분화 전사인자 재조합 단백질을 제조하고, 이 재조합 단백질이 인간 체세포에 만능성(pluripotency)과 자가증식 (self renewal), 미분화 (undifferentiation)성을 유지하게 하여 외부유전자의 삽입 없이 안정하게 역분화를 유도할 수 있음을 확인하고 본 발명을 완성하였다.

따라서 본 발명의 목적은 역분화 전사인자에 세포투과성을 부여하여 이를 세포 내로 고효율로 도입함으로써 체세포의 만능성을 유도하여 유도만능줄기세포를 확립할 수 있는 재조합 단백질을 제공하는 것이다.

상기 목적을 달성하기 위하여, 본 발명은 역분화 전사인자와 거대분자 전송 도메인 (macromolecule transduction domain, MTD)의 융합으로 세포투과성이 부여되어 역분화 전사인자를 세포 내로 고효율로 도입하는 세포투과성 역분화 전사인자 재조합 단백질을 제공한다.

또한 본 발명은 상기 세포투과성 역분화 전사인자 재조합 단백질을 코딩하는 폴리뉴클레오티드를 제공한다.

아울러 본 발명은 상기 폴리뉴클레오티드를 포함하는 발현벡터 및 이 발현벡터로 형질 전환된 형질전환 세균을 제공한다.

또한 본 발명은 상기 형질전환 세균을 배양하는 것을 포함하는 세포투과성 역분화 전사인자 재조합 단백질의 제조방법을 제공한다.

마지막으로 본 발명은 유도만능줄기세포를 안정하고 고효율로 확립할 수 있는 상기 역분화 전사인자 재조합 단백질을 유효성분으로 함유하는 세포투과성 역분화 전사인자를 제공한다.

본 발명에 따른 세포투과성을 갖는 역분화 전사인자 재조합 단백질은 역분화 전사인자를 세포의 핵 내로 고효율로 도입하여 기존에 전사인자 전송의 외부유전자의 삽입으로 인한 문제점을 해결하며 높은 효율로 유도만능 줄기세포를 확립하는데 유용하게 사용될 수 있다.

도 1은 본 발명에 따라 역분화 전사인자 (Nanog, Oct4, Sox2, Klf4, cMyc, Lin28)에 JO-84 및 JO-86 MTDs가 각각 융합된 전사인자 재조합 단백질의 구조를 나타낸 것이고,
도 2는 본 발명의 전사인자 재조합 단백질의 발현을 단백질 발현 유도제인 IPTG의 존재(+) 또는 부재(-) 하에서 조사한 결과이고,
도 3은 본 발명에서 유도만능줄기세포를 만들기 위한 과정이며, 본 발명의 재조합 단백질을 도 3의 과정에 따라 처리하여 인간피부섬유아세포(human dermal fibroblast, HDF)로부터 얻은 유도만능줄기세포의 콜로니(colony)를 알카라인 포스파타아제(alkaline phosphatase) 염색한 결과이고,
도 4는 본 발명의 재조합 단백질을 도 3의 과정에 따라 처리하여 인간피부섬유아세포(human dermal fibroblast, HDF)로부터 얻은 유도만능줄기세포의 콜로니(colony)를 줄기세포 핵에서 특이적으로 발현하는 유전자(Oct4 ＆Nanog)로 염색한 결과이고,
도 5는 본 발명의 재조합 단백질을 도 3의 과정에 따라 처리하여 인간피부섬유아세포(human dermal fibroblast, HDF)로부터 얻은 유도만능줄기세포의 줄기세포 표면에서 발현하는 특이적 마커 (stage-specific embryonic antigen-3, SSEA-3)를 유세포 분석기를 이용하여 확인한 결과이고,
도 6은 본 발명의 재조합 단백질을 도 3의 과정에 따라 처리하여 인간피부섬유아세포(human dermal fibroblast, HDF)로부터 얻은 유도만능줄기세포의 RNA를 역전사(reverse transcription) PCR를 이용하여 줄기세포 특이 유전자가 발현됨을 확인한 결과이고,
도 7은 본 발명에서 유도만능줄기세포를 만들기 위한 과정이며, 본 발명의 재조합 단백질을 도 7의 과정에 따라 처리하여 인간피부섬유아세포(human dermal fibroblast, HDF)로부터 얻은 유도만능줄기세포의 콜로니(colony)를 현미경으로 관찰하여 카메라로 찍은 사진이고,
도 8은 본 발명의 재조합 단백질을 도 7의 과정에 따라 처리하여 인간피부섬유아세포(human dermal fibroblast, HDF)로부터 얻은 유도만능줄기세포의 콜로니(colony)를 줄기세포 핵에서 특이적으로 발현하는 유전자(Oct4 ＆Nanog)로 염색한 결과이고,
도 9는 본 발명의 재조합 단백질을 도 7의 과정에 따라 처리하여 인간피부섬유아세포(human dermal fibroblast, HDF)로부터 얻은 유도만능줄기세포의 줄기세포 표면에서 발현하는 특이적 마커 SSEA-3 (Stage-Specific Embryonic Antigen-3)와 TRA-1-60 (Tumor Rejection Antigen-1-60)를 유세포 분석기를 이용하여 확인한 결과이고,
도 10 은 본 발명에 따라 역분화 전사인자 (Nanog, Oct4, Sox2, Klf4, cMyc, Lin28)에 JO-10, JO-52, JO-132, JO-145, JO-173 및 JO-181 MTDs 가 각각 융합된 전사인자 재조합 단백질의 구조를 나타낸 것이고,
도 11은 본 발명의 전사인자 재조합 단백질의 발현을 최적화 하기 위하여 codon optimization과정을 거친 재조합 단백질의 발현을 발현 유도제인 IPTG의 존재(+) 또는 부재(-) 하에서 조사한 결과이다.

본 발명은 역분화 전사인자 Nanog, Oct4, Sox2, Klf4, cMyc, Lin28 과 거대분자 전달 도메인 (MTD)의 융합으로 세포투과성이 부여되어 Nanog, Oct4, Sox2, Klf4, cMyc, Lin28 을 세포 내로 고효율로 도입할 수 있는 세포투과성 Nanog, Oct4, Sox2, Klf4, cMyc, Lin28 재조합 단백질 (CP-Nanog, CP-Oct4, CP-Sox2, CP-Klf4, CP-cMyc, CP-Lin28) 및 이를 코딩하는 폴리뉴클레오티드를 제공한다.

본 발명은 역분화 전사인자 Nanog, Oct4, Sox2, Klf4, cMyc, Lin28과 거대분자 전달 도메인 (MTD)의 융합으로 세포투과성이 부여된 단백질을 사용하였다.

상기에서 세포핵 배치 서열(NLS)에 전장형태의 Nanog가 융합된 재조합 단백질로서

NLS-Nanog(NNanog)는 서열번호: 20의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 19의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-84 MTD(MTD₈₄)를 이용하여 제조된 전장 형태의 Nanog 재조합 단백질로서,

NLS-MTD₈₄-Nanog(NM₈₄Nanog)는 서열번호: 22 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 21 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Nanog-MTD₈₄ (NNanogM₈₄)은 서열번호: 24 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 23 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₄-Nanog-MTD₈₄(NM₈₄NanogM₈₄)은 서열번호: 26 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 25 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-86 MTD(MTD₈₆)를 이용하여 제조된 전장 형태의 Nanog 재조합 단백질로서,

NLS-MTD₈₆-Nanog(NM₈₆Nanog)는 서열번호: 28 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 27 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Nanog-MTD₈₆ (NNanogM₈₆)은 서열번호: 30 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 29 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₆-Nanog-MTD₈₆(NM₈₆NanogM₈₆)은 서열번호: 32 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 31 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

또한, 본 발명에서는 분리, 정제의 용이성을 위하여 상기 Nanog 재조합 단백질에 히스티딘 표지(His-tag)가 추가로 포함된 형태도 사용된다.

His-NLS-Nanog(HNNanog)는 서열번호: 34 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 33 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다. 상기에서 JO-84 MTD(MTD₈₄)를 이용하여 제조된 전장 형태의 Nanog 재조합 단백질로서,

His-NLS-MTD₈₄-Nanog(HNM₈₄Nanog)은 서열번호: 36 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 35 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Nanog-MTD₈₄(HNNanogM₈₄)은 서열번호: 38 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 37 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₄-Nanog-MTD₈₄(HNM₈₄NanogM₈₄)은 서열번호: 40 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 39 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₆-Nanog(HNM₈₆Nanog)은 서열번호: 42 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 41 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Nanog-MTD₈₆(HNNanogM₈₆)은 서열번호: 44 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 43 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₆-Nanog-MTD₈₆(HNM₈₆NanogM₈₆)은 서열번호: 46 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 45 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고,

His-NLS-MTD₁₀-Nanog(HNM₁₀Nanog)은 서열번호: 237 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 236 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 세포핵 배치 서열(NLS)에 전장형태의 Oct4가 융합된 재조합 단백질로서

NLS-Oct4(NOct4)는 서열번호: 48의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 47의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-84 MTD(MTD₈₄)를 이용하여 제조된 전장 형태의 Oct4 재조합 단백질로서,

NLS-MTD₈₄-Oct4(NM₈₄Oct4)는 서열번호: 50 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 49 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Oct4-MTD₈₄ (NOct4M₈₄)은 서열번호: 52 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 51 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₄-Oct4-MTD₈₄(NM₈₄Oct4M₈₄)은 서열번호: 54 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 53 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-86 MTD(MTD₈₆)를 이용하여 제조된 전장 형태의 Oct4 재조합 단백질로서,

NLS-MTD₈₆-Oct4(NM₈₆Oct4)는 서열번호: 56 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 55 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Oct4MTD₈₆ (NOct4M₈₆)은 서열번호: 58 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 57 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₆-Oct4-MTD₈₆(NM₈₆Oct4M₈₆)은 서열번호: 60 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 59 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

또한, 본 발명에서는 분리, 정제의 용이성을 위하여 상기 Oct4 재조합 단백질에 히스티딘 표지(His-tag)가 추가로 포함된 형태도 사용된다.

His-NLS-Oct4(HNOct4)는 서열번호: 62 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 61 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₄-Oct4(HNM₈₄Oct4)은 서열번호: 64 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 63 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Oct4-MTD₈₄(HNOct4M₈₄)은 서열번호: 66 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 65 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₄-Oct4-MTD₈₄(HNM₈₄Oct4M₈₄)은 서열번호: 68 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 67 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₆-Oct4(HNM₈₆Oct4)은 서열번호: 70 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 69 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Oct4-MTD₈₆(HNOct4M₈₆)은 서열번호: 72 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 71 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₆-Oct4-MTD₈₆(HNM₈₆Oct4M₈₆)은 서열번호: 74 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 73 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고,

His-NLS-MTD₅₂-Oct4(HNM₅₂Oct4)은 서열번호: 240 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 239 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 세포핵 배치 서열(NLS)에 전장형태의 Sox2가 융합된 재조합 단백질로서

NLS-Sox2(NSox2)는 서열번호: 76의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 75의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-84 MTD(MTD₈₄)를 이용하여 제조된 전장 형태의 Sox2 재조합 단백질로서,

NLS-MTD₈₄-Sox2(NM₈₄Sox2)는 서열번호: 78 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 77 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Sox2-MTD₈₄ (NSox2M₈₄)은 서열번호: 80 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 79 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₄-Sox2-MTD₈₄(NM₈₄Sox2M₈₄)은 서열번호: 82 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 81 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-86 MTD(MTD₈₆)를 이용하여 제조된 전장 형태의 Sox2 재조합 단백질로서,

NLS-MTD₈₆-Sox2(NM₈₆Sox2)는 서열번호: 84 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 83 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Sox2-MTD₈₆ (NSox2M₈₆)은 서열번호: 86 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 85 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₆-Sox2-MTD₈₆(NM₈₆Sox2M₈₆)은 서열번호: 88 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 87 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

또한, 본 발명에서는 분리, 정제의 용이성을 위하여 상기 Sox2 재조합 단백질에 히스티딘 표지(His-tag)가 추가로 포함된 형태도 사용된다.

His-NLS-Sox2(HNSox2)는 서열번호: 90 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 89 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₄-Sox2(HNM₈₄Sox2)은 서열번호: 92 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 91 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Sox2-MTD₈₄(HNSox2M₈₄)은 서열번호: 94 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 93 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₄-Sox2-MTD₈₄(HNM₈₄Sox2M₈₄)은 서열번호: 96 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 95 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₆-Sox2(HNM₈₆Sox2)은 서열번호: 98 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 97 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Sox2-MTD₈₆(HNSox2M₈₆)은 서열번호: 100 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 99 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₆-Sox2-MTD₈₆(HNM₈₆Sox2M₈₆)은 서열번호: 102 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 101 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고,

His-NLS-MTD₁₈₁-Sox2(HNM₁₈₁Sox2)은 서열번호: 243 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 242 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 세포핵 배치 서열(NLS)에 전장형태의 Klf4가 융합된 재조합 단백질로서

NLS-Klf4(NKlf4)는 서열번호: 104의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 103의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-84 MTD(MTD₈₄)를 이용하여 제조된 전장 형태의 Klf4 재조합 단백질로서,

NLS-MTD₈₄-Klf4(NM₈₄Klf4)는 서열번호: 106 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 105 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Klf4-MTD₈₄(NKlf4M₈₄)은 서열번호: 108 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 107 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₄-Klf4-MTD₈₄(NM₈₄Klf4M₈₄)은 서열번호: 110 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 109 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-86 MTD(MTD₈₆)를 이용하여 제조된 전장 형태의 Klf4 재조합 단백질로서,

NLS-MTD₈₆-Klf4(NM₈₆Klf4)는 서열번호: 112 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 111 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Klf4-MTD₈₆ (NKlf4M₈₆)은 서열번호: 114 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 113 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₆-Klf4-MTD₈₆(NM₈₆Klf4M₈₆)은 서열번호: 116 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 115 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

또한, 본 발명에서는 분리, 정제의 용이성을 위하여 상기 Klf4 재조합 단백질에 히스티딘 표지(His-tag)가 추가로 포함된 형태도 사용된다.

His-NLS-Klf4(HNKlf4)는 서열번호: 118의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 117의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₄-Klf4(HNM₈₄Klf4)은 서열번호: 120 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 119 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Klf4-MTD₈₄(HNKlf4M₈₄)은 서열번호: 122 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 121 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₄-Klf4-MTD₈₄(HNM₈₄Klf4M₈₄)은 서열번호: 124 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 123 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₆-Klf4(HNM₈₆Klf4)은 서열번호: 126 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 125 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Klf4-MTD₈₆(HNKlf4M₈₆)은 서열번호: 128 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 127 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₆-Klf4-MTD₈₆(HNM₈₆Klf4M₈₆)은 서열번호: 130 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 129 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고

His-NLS-MTD₁₇₃-Klf4(HNM₁₇₃Klf4)은 서열번호: 246 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 245 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

상기에서 세포핵 배치 서열(NLS)에 전장형태의 cMyc이 융합된 재조합 단백질로서

NLS-cMyc(NcMyc)는 서열번호: 132의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 131의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-84 MTD(MTD₈₄)를 이용하여 제조된 전장 형태의 cMyc 재조합 단백질로서,

NLS-MTD₈₄-cMyc(NM₈₄cMyc)는 서열번호: 134 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 133 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-cMyc-MTD₈₄(NcMycM₈₄)은 서열번호: 136 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 135 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₄-cMyc-MTD₈₄(NM₈₄cMycM₈₄)은 서열번호: 138 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 137 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-86 MTD(MTD₈₆)를 이용하여 제조된 전장 형태의 cMyc 재조합 단백질로서,

NLS-MTD₈₆-cMyc(NM₈₆cMyc)는 서열번호: 140 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 139 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-cMyc-MTD₈₆ (NcMycM₈₆)은 서열번호: 142 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 141 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₆-cMyc-MTD₈₆(NM₈₆cMycM₈₆)은 서열번호: 144 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 143 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

또한, 본 발명에서는 분리, 정제의 용이성을 위하여 상기 cMyc 재조합 단백질에 히스티딘 표지(His-tag)가 추가로 포함된 형태도 사용된다.

His-NLS-cMyc(HNcMyc)는 서열번호: 146의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 145의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₄-cMyc(HNM₈₄cMyc)은 서열번호: 148 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 147 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-cMyc-MTD₈₄(HNcMycM₈₄)은 서열번호: 150 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 149 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₄-cMyc-MTD₈₄(HNM₈₄cMycM₈₄)은 서열번호: 152 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 151 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₆-cMyc(HNM₈₆cMyc)은 서열번호: 154 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 153 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-cMyc-MTD₈₆(HNcMycM₈₆)은 서열번호: 156 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 155 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₆-cMyc-MTD₈₆(HNM₈₆cMycM₈₆)은 서열번호: 158 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 157 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고,

His-NLS-MTD₁₄₅-cMyc(HNM₁₄₅cMyc)은 서열번호: 249 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 248 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 세포핵 배치 서열(NLS)에 전장형태의 Lin28이 융합된 재조합 단백질로서

NLS-Lin28(NLin28)는 서열번호: 160의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 159의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-84 MTD(MTD₈₄)를 이용하여 제조된 전장 형태의 Lin28 재조합 단백질로서,

NLS-MTD₈₄-Lin28(NM₈₄Lin28)는 서열번호: 162 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 161 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Lin28-MTD₈₄(NLin28M₈₄)은 서열번호: 164 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 163 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₄-Lin28-MTD₈₄(NM₈₄Lin28M₈₄)은 서열번호: 166 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 165 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

상기에서 JO-86 MTD(MTD₈₆)를 이용하여 제조된 전장 형태의 Lin28 재조합 단백질로서,

NLS-MTD₈₆-Lin28(NM₈₆Lin28)는 서열번호: 168 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 167 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-Lin28-MTD₈₆ (NLin28M₈₆)은 서열번호: 170 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 169 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

NLS-MTD₈₆-Lin28-MTD₈₆(NM₈₆Lin28M₈₆)은 서열번호: 172 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 171 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

또한, 본 발명에서는 분리, 정제의 용이성을 위하여 상기 Lin28 재조합 단백질에 히스티딘 표지(His-tag)가 추가로 포함된 형태도 사용된다.

His-NLS-Lin28(HNLin28)는 서열번호: 174의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 173의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₄-Lin28(HNM₈₄Lin28)은 서열번호: 176 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 175 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Lin28-MTD₈₄(HNLin28M₈₄)은 서열번호: 178 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 177 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₄-Lin28-MTD₈₄(HNM₈₄Lin28M₈₄)은 서열번호: 180 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 179 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

His-NLS-MTD₈₆-Lin28(HNM₈₆Lin28)은 서열번호: 182 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 181 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-Lin28-MTD₈₆(HNLin28M₈₆)은 서열번호: 184 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 183 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고;

His-NLS-MTD₈₆-Lin28-MTD₈₆(HNM₈₆Lin28M₈₆)은 서열번호: 186 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 185 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩되고,

His-NLS-MTD₁₃₂-Lin28(HNM₁₃₂Lin28)은 서열번호: 252 의 아미노산 서열을 가지며, 이는 예를 들면, 서열번호: 251 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

본 발명의 특징은 거대분자 세포 내 전달기술 (MITT)을 이용하여 세포 내로의 도입이 용이하지 않은 거대분자인 역분화 전사인자에 특정한 거대분자 전달 도메인 (이하, "MTD"로 약칭함)을 융합시켜 세포투과성을 부여함으로써 역분화 전사인자를 세포 내로 고효율로 전달하는 것이다. 이때 거대분자 전달 도메인은 역분화 전사인자의 한쪽 말단에만 융합되거나, 또는 그의 양쪽 말단 모두에 융합될 수 있다. 분비 단백질 (secreted protein) 유래 소수성 펩티드인 MTD를 이용한 세포 내 전달기술은 단백질의 처리 만으로 세포막을 투과하여 핵내로 전송되어질 수 있다.

이러한 효과는 체세포에서 발현되지 않는 역분화 전사인자를 세포내 전송하여 기존 바이러스, 플라스미드와 같은 외부유전체의 삽입없이 안전하게 유도만능줄기세포를 확립 할 수 있게 한다.

본 발명에서는 역분화 전사인자에 융합될 수 있는 거대분자 전달 도메인 (MTD)으로서 세포 내로 거대분자의 전달을 가능케 하는 펩티드 도메인에 상기 역분화 전사인자를 융합하여 세포투과성을 갖는 역분화 전사인자 재조합 단백질을 개발 하였다.

본 발명에서 "세포투과성 역분화 전사인자 재조합 단백질" 이란 거대분자 전달 도메인과 역분화 전사인자를 포함하며, 이들의 유전적 융합이나 화학적 결합으로 형성된 공유결합 복합체를 의미한다. 여기서 "유전적 융합" 이란 단백질을 코딩하는 DNA 서열의 유전적 발현을 통해서 형성된 선형의 공유결합으로 이루어진 연결을 의미한다.

체세포의 역분화능을 가진 전사인자 Nanog 는 서열번호: 1 의 염기서열 및 서열번호: 2 아미노산 서열을 갖는 전사인자로서 배아의 만능성을 유지시켜주며 내배엽으로의 분화를 억제하는 전사인자다. 역분화 전사인자 Nanog 는 서열번호: 2 의 아미노산 서열에서 1 번부터 306 번까지의 아미노산을 포함하는 도메인으로 구성된다.

체세포의 역분화능을 가진 전사인자 Oct4는 서열번호: 3의 염기서열 및 서열번호: 4 아미노산 서열을 갖는 전사인자로서 줄기세포의 만능성을 유지시켜주며 자기 재생산능을 조절하는 전사인자다. 역분화 전사인자 Oct4는 서열번호: 4의 아미노산 서열에서 1번부터 361번까지의 아미노산을 포함하는 도메인으로 구성된다.

체세포의 역분화능을 가진 전사인자 Sox2 는 서열번호: 5 의 염기서열 및 서열번호: 6 아미노산 서열을 갖는 전사인자로서 줄기세포의 만능성을 유지시켜주며 줄기세포의 미분화능을 조절하는 전사인자다. 역분화 전사인자 Sox2 는 서열번호: 6 의 아미노산 서열에서 1 번부터 318 번까지의 아미노산을 포함하는 도메인으로 구성된다.

체세포의 역분화능을 가진 전사인자 Klf4 는 서열번호: 7 의 염기서열 및 서열번호: 8 아미노산 서열을 갖는 전사인자로서 세포의 성장억제와 세포주기를 조절하여 역분화를 유도하는 전사인자다. 역분화 전사인자 Klf4 는 서열번호: 8 의 아미노산 서열에서 1 번부터 471 번까지의 아미노산을 포함하는 도메인으로 구성된다.

체세포의 역분화능을 가진 전사인자 cMyc 는 서열번호: 9 의 염기서열 및 서열번호: 10 아미노산 서열을 갖는 전사인자로서 세포의 성장, 증식, 세포 예정사와 세포주기를 조절하여 역분화를 유도하는 전사인자다. cMyc 는 역분화 전사인자 서열번호: 10 의 아미노산 서열에서 1 번부터 456 번까지의 아미노산을 포함하는 도메인으로 구성된다.

체세포의 역분화능을 가진 전사인자 Lin28 은 서열번호: 11 의 염기서열 및 서열번호: 12 아미노산 서열을 갖는 전사인자로서 배아줄기세포의 분화와 증식을 조절하는 전사인자다. 역분화 전사인자 Lin28 은 서열번호: 12 의 아미노산 서열에서 1 번부터 210 번까지의 아미노산을 포함하는 도메인으로 구성된다.

역분화 전사인자 6 종에 융합될 수 있는 다른 거대분자 전달 도메인으로는 서열번호: 15 의 염기서열 및 서열번호: 16 의 아미노산 서열을 갖는 세포투과성을 갖는 폴리펩티드가 사용될 수 있다.

역분화 전사인자 6 종에 융합될 수 있는 다른 거대분자 전달 도메인으로는 서열번호: 17 의 염기서열 및 서열번호: 18 의 아미노산 서열을 갖는 세포투과성을 갖는 폴리펩티드가 사용될 수 있다.

역분화 전사인자 Nanog 에 융합될 수 있는 다른 거대분자 전달 도메인으로는 서열번호: 223 의 염기서열 및 서열번호: 224 의 아미노산 서열을 갖는 세포투과성을 갖는 폴리펩티드가 사용될 수 있다.

역분화 전사인자 Oct4 에 융합될 수 있는 다른 거대분자 전달 도메인으로는 서열번호: 225 의 염기서열 및 서열번호: 226 의 아미노산 서열을 갖는 세포투과성을 갖는 폴리펩티드가 사용될 수 있다.

역분화 전사인자 Sox2 에 융합될 수 있는 다른 거대분자 전달 도메인으로는 서열번호: 233 의 염기서열 및 서열번호: 234 의 아미노산 서열을 갖는 세포투과성을 갖는 폴리펩티드가 사용될 수 있다.

역분화 전사인자 Klf4 에 융합될 수 있는 다른 거대분자 전달 도메인으로는 서열번호: 231 의 염기서열 및 서열번호: 232 의 아미노산 서열을 갖는 세포투과성을 갖는 폴리펩티드가 사용될 수 있다.

역분화 전사인자 cMyc 에 융합될 수 있는 다른 거대분자 전달 도메인으로는 서열번호: 229 의 염기서열 및 서열번호: 230 의 아미노산 서열을 갖는 세포투과성을 갖는 폴리펩티드가 사용될 수 있다.

역분화 전사인자 Lin28 에 융합될 수 있는 다른 거대분자 전달 도메인으로는 서열번호: 227 의 염기서열 및 서열번호: 228 의 아미노산 서열을 갖는 세포투과성을 갖는 폴리펩티드가 사용될 수 있다.

상기 서열번호: 16, 18, 224, 226, 228, 230, 232, 234 으로 기재되는 아미노산 서열 중의 어느 하나를 포함하는 거대분자 전달 도메인은 세포막을 관통하여 폴리펩티드, 단백질 도메인, 또는 전장 단백질을 포함하는 생물학적 활성 분자의 세포 내로의 유입을 매개할 수 있는 세포투과성 폴리펩티드이다. 본 발명에 따른 거대분자 전달 도메인은 N-말단 영역, 소수성 영역 및 C-말단의 분비 단백질 절단 부위 (secreted protein cleavage site)의 3 부분으로 이루어진 시그널 펩티드 (signal peptides)에서 헬릭스 (helix)를 형성해 세포막 표적 (targeting) 활성을 부여하는 소수성 영역을 갖도록 고안된 것이다. 이러한 거대분자 전달 도메인은 세포에 손상을 주지 않으면서 직접 세포막을 관통함으로써 표적 단백질을 세포 내로 이동시켜 목적하는 기능을 발휘하게 할 수 있다.

본 발명에 따라 역분화 전사인자와 융합될 수 있는 서열번호: 16, 18, 224, 226, 228, 230, 232, 234 로 기재되는 아미노산 서열을 갖는 거대분자 전달 도메인을 하기 표 1 에 나타내었다.

본 발명의 세포투과성을 갖는 역분화 전사인자 재조합 단백질은 거대분자 전달 도메인으로 상기 8 종의 MTDs 중의 어느 하나가 종양 역분화 전사인자의 한쪽 또는 양쪽 말단에 융합되고, 필요에 따라, 이 융합 컨스트럭트의 한쪽 말단에 SV40 거대 T 항원 (SV40 large T antigen) 유래 세포핵 배치 서열 (nuclear localization sequence: NLS)과 용이한 정제를 위한 히스티딘-표지 (histidine-tag, His-Tag) 친화성 도메인이 융합되어 있는 구조를 가질 수 있다.

역분화 전사인자에 융합될 수 있는 세포핵 배치서열로는 서열번호: 13 의 염기서열 및 서열번호: 14 의 아미노산 서열을 갖는 폴리펩티드가 사용될 수 있으나, 이에 제한 되지 않으며, 당업계에 잘 알려져 있는 다른 세포핵 배치서열도 사용될 수 있다.

본 발명의 일 실시양태에서는, 역분화 전사인자에 융합될 수 있는 거대분자 전달 도메인으로 스트렙토마이스 코엘리콜러(Streptomyces coelicolor) 로부터 유래된 서열번호: 16 의 아미노산 서열을 갖는 JO-84 MTD (이하, "MTD₈₄"로 약칭함), 마이코박테리움 보바이스(Mycobacterium bovis)로 부터 유래된 분비 단백질 (secreted protein)인 서열번호: 18 의 아미노산 서열을 갖는 JO-86 MTD (이하, "MTD₈₆"로 약칭함), 스트렙토마이스 코엘리콜러(Streptomyces coelicolor) 로부터 유래된 서열번호: 224 의 아미노산 서열을 갖는 JO-10 MTD (이하, "MTD₁₀"로 약칭함), 호모 사피엔스(Homo Sapiens)로부터 유래된 서열번호: 226 의 아미노산 서열을 갖는 JO-52 MTD (이하, "MTD₅₂"로 약칭함), 스트렙토마이스 코엘리콜러(Streptomyces coelicolor) 로부터 유래된 서열번호: 228 의 아미노산 서열을 갖는 JO-132 MTD (이하, "MTD₁₃₂"로 약칭함), 호모 사피엔스 엘라스틴 (Homo Sapiens Elastin)으로부터 유래된 서열번호: 230 의 아미노산 서열을 갖는 JO-145 MTD (이하, "MTD₁₄₅"로 약칭함), 스트렙토마이스 코엘리콜러(Streptomyces coelicolor) 로부터 유래된 서열번호: 232 의 아미노산 서열을 갖는 JO-173 MTD (이하, "MTD₁₇₃"로 약칭함), 니제리아 메닌티디스(Neisseria meningitidis) 로부터 유래된 서열번호: 234 의 아미노산 서열을 갖는 JO-181 MTD (이하, "MTD₁₈₁"로 약칭함),를 사용한다.

본 발명의 바람직한 실시양태에서는, 거대분자 전달 도메인으로 JO-84 MTD 또는 JO-86 MTD 를 이용한 세포투과성 역분화 전사인자 재조합 단백질로서 이들 각각에 대해 2 개의 전장 형태 (full-length forms)를 고안한다.

도 1, 10 을 참고로 하면, 본 발명에 따른 6 종의 역분화 전사인자 재조합단백질중 Nanog 는

1) Nanog 의 N-말단에 His-NLS 와 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Nanog(HNM₈₄Nanog),

2) Nanog 의 N-말단에 His-NLS 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-Nanog-MTD₈₄(HNNanogM₈₄),

3) Nanog 의 N-말단에 His-NLS-JO-84 MTD 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Nanog-MTD₈₄(HNM₈₄NanogM₈₄),

4) Nanog 의 N-말단에 His-NLS 와 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Nanog(HNM₈₆Nanog),

5) Nanog 의 N-말단에 His-NLS 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-Nanog-MTD₈₆(HNNanogM₈₆),

6) Nanog 의 N-말단에 His-NLS-JO-86 MTD 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Nanog-MTD₈₆(HNM₈₆NanogM₈₆)의 형태 일 수도 있고,

7) Nanog 의 N-말단에 His-NLS-JO-10 MTD 가 융합된 His-NLS-MTD₁₀-Nanog (HNM₁₀Nanog)의 형태 일 수도 있고,

또는

상기 1) 내지 7)의 재조합 단백질에서 히스티딘 표지 (His-tag)가 포함되지 않은 형태일 수도 있다.

도 1, 10 을 참고로 하면, 본 발명에 따른 6 종의 역분화 전사인자 재조합 단백질중 Oct4 는

1) Oct4 의 N-말단에 His-NLS 와 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Oct4(HNM₈₄Oct4),

2) Oct4 의 N-말단에 His-NLS 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-Oct4-MTD₈₄(HNOct4M₈₄),

3) Oct4 의 N-말단에 His-NLS-JO-84 MTD 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Oct4-MTD₈₄(HNM₈₄Oct4M₈₄),

4) Oct4 의 N-말단에 His-NLS 와 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Oct4(HNM₈₆Oct4),

5) Oct4 의 N-말단에 His-NLS 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-Oct4-MTD₈₆(HNOct4M₈₆),

6) Oct4 의 N-말단에 His-NLS-JO-86 MTD 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Oct4-MTD₈₆(HNM₈₆Oct4M₈₆)의 형태일 수도 있고,

7) Oct4 의 N-말단에 His-NLS 와 JO-52 MTD 가 융합된 His-NLS-MTD₅₂-Oct4(HNM₅₂Oct4),

또는

도 1, 10 을 참고로 하면, 본 발명에 따른 6 종의 역분화 전사인자 재조합단백질중 Sox2 는

1) Sox2 의 N-말단에 His-NLS 와 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Sox2(HNM₈₄Sox2),

2) Sox2 의 N-말단에 His-NLS 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-Sox2-MTD₈₄(HNSox2M₈₄),

3) Sox2 의 N-말단에 His-NLS-JO-84 MTD 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Sox2-MTD₈₄(HNM₈₄Sox2M₈₄),

4) Sox2 의 N-말단에 His-NLS 와 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Sox2(HNM₈₆Sox2),

5) Sox2 의 N-말단에 His-NLS 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-Sox2-MTD₈₆(HNSox2M₈₆),

6) Sox2 의 N-말단에 His-NLS-JO-86 MTD 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Sox2-MTD₈₆(HNM₈₆Sox2M₈₆)의 형태일 수도 있고,

7) Sox2 의 N-말단에 His-NLS 와 JO-181 MTD 가 융합된 His-NLS-MTD₁₈₁-Sox2(HNM₁₈₁Sox2),

또는

도 1, 10 을 참고로 하면, 본 발명에 따른 6 종의 역분화 전사인자 재조합단백질중 Klf4 는

1) Klf4 의 N-말단에 His-NLS 와 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Klf4(HNM₈₄Klf4),

2) Klf4 의 N-말단에 His-NLS 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-Klf4-MTD₈₄(HNKlf4M₈₄),

3) Klf4 의 N-말단에 His-NLS-JO-84 MTD 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Klf4-MTD₈₄(HNM₈₄Klf4M₈₄),

4) Klf4 의 N-말단에 His-NLS 와 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Klf4(HNM₈₆Klf4),

5) Klf4 의 N-말단에 His-NLS 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-Klf4-MTD₈₆(HNKlf4M₈₆),

6) Klf4 의 N-말단에 His-NLS-JO-86 MTD 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Klf4-MTD₈₆(HNM₈₆Klf4M₈₆)의 형태일 수도 있고,

7) Klf4 의 N-말단에 His-NLS 와 JO-173 MTD 가 융합된 His-NLS-MTD₁₇₃-Klf4(HNM₁₇₃Klf4),

또는

도 1, 10 을 참고로 하면, 본 발명에 따른 6 종의 역분화 전사인자 재조합단백질중 cMyc 는

1) cMyc 의 N-말단에 His-NLS 와 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-cMyC(HNM₈₄cMyc),

2) cMyc 의 N-말단에 His-NLS 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-cMyc-MTD₈₄(HNcMycM₈₄),

3) cMyc 의 N-말단에 His-NLS-JO-84 MTD 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-cMyc-MTD₈₄(HNM₈₄cMycM₈₄),

4) cMyc 의 N-말단에 His-NLS 와 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-cMyc(HNM₈₆cMyc),

5) cMyc 의 N-말단에 His-NLS 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-cMyc-MTD₈₆(HNcMycM₈₆),

6) cMyc 의 N-말단에 His-NLS-JO-86 MTD 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-cMyc-MTD₈₆(HNM₈₆cMycM₈₆)의 형태일 수도 있고,

7) cMyc 의 N-말단에 His-NLS 와 JO-145 MTD 가 융합된 His-NLS-MTD₁₄₅-cMyc(HNM₁₄₅cMyc),

또는

도 1, 10 을 참고로 하면, 본 발명에 따른 6 종의 역분화 전사인자 재조합단백질중 Lin28 는

1) Lin28 의 N-말단에 His-NLS 와 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Lin28(HNM₈₄Lin28),

2) Lin28 의 N-말단에 His-NLS 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-Lin28-MTD₈₄(HNLin28M₈₄),

3) Lin28 의 N-말단에 His-NLS-JO-84 MTD 와 C-말단에 JO-84 MTD 가 융합된 His-NLS-MTD₈₄-Lin28-MTD₈₄(HNM₈₄Lin28M₈₄),

4) Lin28 의 N-말단에 His-NLS 와 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Lin28(HNM₈₆Lin28),

5) Lin28 의 N-말단에 His-NLS 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-Lin28-MTD₈₆(HNLin28M₈₆),

6) Lin28 의 N-말단에 His-NLS-JO-86 MTD 와 C-말단에 JO-86 MTD 가 융합된 His-NLS-MTD₈₆-Lin28-MTD₈₆(HNM₈₆Lin28M₈₆)의 형태일 수도 있고,

7) Lin28 의 N-말단에 His-NLS 와 JO-132 MTD 가 융합된 His-NLS-MTD₁₃₂-Lin28(HNM₁₃₂Lin28),

또는

본 발명에서는 상기 세포투과성 6 종의 역분화 전사인자 재조합 단백질중 Nanog 의 대조군으로서 MTD 가 융합되지 않고 히스티딘 표지와 NLS 만이 융합된 재조합 단백질 His-NLS-Nanog(HNNanog)을 제조한다. 이 대조군 단백질은 서열번호: 34 의 아미노산 서열을 가지며, 이는 서열번호: 33 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

본 발명에서는 상기 세포투과성 6 종의 역분화 전사인자 재조합 단백질중 Oct4 의 대조군으로서 MTD 가 융합되지 않고 히스티딘 표지와 NLS 만이 융합된 재조합 단백질 His-NLS-Oct4(HNOct4)을 제조한다. 이 대조군 단백질은 서열번호: 62 의 아미노산 서열을 가지며, 이는 서열번호: 61 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

본 발명에서는 상기 세포투과성 6 종의 역분화 전사인자 재조합 단백질중 Sox2 의 대조군으로서 MTD 가 융합되지 않고 히스티딘 표지와 NLS 만이 융합된 재조합 단백질 His-NLS-Sox2(HNSox2)을 제조한다. 이 대조군 단백질은 서열번호: 90 의 아미노산 서열을 가지며, 이는 서열번호: 89 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

본 발명에서는 상기 세포투과성 6 종의 역분화 전사인자 재조합 단백질중 Klf4 의 대조군으로서 MTD 가 융합되지 않고 히스티딘 표지와 NLS 만이 융합된 재조합 단백질 His-NLS-Klf4(HNKlf4)을 제조한다. 이 대조군 단백질은 서열번호: 118 의 아미노산 서열을 가지며, 이는 서열번호: 117 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

본 발명에서는 상기 세포투과성 6 종의 역분화 전사인자 재조합 단백질중 cMyc 의 대조군으로서 MTD 가 융합되지 않고 히스티딘 표지와 NLS 만이 융합된 재조합 단백질 His-NLS-cMyc(HNcMyc)을 제조한다. 이 대조군 단백질은 서열번호: 146 의 아미노산 서열을 가지며, 이는 서열번호: 145 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

본 발명에서는 상기 세포투과성 6 종의 역분화 전사인자 재조합 단백질중 Lin28 의 대조군으로서 MTD 가 융합되지 않고 히스티딘 표지와 NLS 만이 융합된 재조합 단백질 His-NLS-Lin28(HNLin28)을 제조한다. 이 대조군 단백질은 서열번호: 174 의 아미노산 서열을 가지며, 이는 서열번호: 173 의 염기서열을 갖는 폴리뉴클레오티드에 의해 코딩된다.

나아가, 본 발명은 상기 세포투과성 역분화 전사인자 재조합 단백질을 코딩하는 폴리뉴클레오티드를 포함하는 재조합 발현벡터 및 이 재조합 발현벡터로 형질전환된 형질전환 세균을 제공한다.

본 발명에서 "재조합 발현벡터"란 적당한 숙주세포에서 목적 단백질 또는 목적 RNA 를 발현할 수 있는 벡터로서, 유전자 삽입물이 발현되도록 작동가능하게 연결된 필수적인 조절요소를 포함하는 유전자 제작물을 말한다.

본 발명에서 용어, "작동가능하게 연결된 (operably linked)"은 일반적 기능을 수행하도록 핵산 발현조절 서열과 목적하는 단백질 또는 RNA 를 코딩하는 핵산 서열이 기능적으로 연결 (functional linkage)되어 있는 것을 말한다. 예를 들어 프로모터와 단백질 또는 RNA 를 코딩하는 핵산 서열이 작동가능하게 연결되어 코딩하는 핵산 서열의 발현에 영향을 미칠 수 있다. 재조합 발현벡터와의 작동적 연결은 당해 기술 분야에 잘 알려진 유전자 재조합 기술을 이용하여 달성할 수 있으며, 부위-특이적 DNA 절단 및 연결은 당해 기술 분야에 일반적으로 알려진 효소 등을 사용한다.

본 발명에 사용가능한 발현벡터는 플라스미드 벡터, 코스미드 (cosmid) 벡터, 박테리오파아지 벡터, 바이러스 벡터 등을 포함하나 이에 제한되지 않는다. 적합한 발현벡터는 프로모터 (promoter), 오퍼레이터 (operator), 개시코돈 (initiation codon), 종결코돈 (termination codon), 폴리아데닐화 신호 (polyadenylation signal), 인핸서 (enhancer)와 같은 발현 조절서열 외에도, 막 표적화 또는 분비를 위한 신호서열 (signal sequence) 또는 리더서열 (leader sequence)을 포함하여 목적에 따라 다양하게 제조될 수 있다. 발현벡터의 프로모터는 구성적 (constitutive) 또는 유도성 (inducible)일 수 있다. 또한 발현벡터는 벡터를 함유하는 숙주세포를 선택하기 위한 선택마커를 포함하고, 복제가 가능한 발현벡터인 경우 복제 기원을 포함한다.

이와 같이 제조된 본 발명의 재조합 발현벡터는 예를 들면, pET28a(+)-HNM₈₆Nanog, pET28a(+)-HNM₈₆Oct4, pET28a(+)-HNM₈₆Sox2M₈₆, pET28a(+)-HNM₈₆Klf4M₈₆, pET28a(+)-HNM₈₆cMyc 일 수 있다.상기 재조합 발현벡터 pET28a(+)-HNM₈₆Nanog, pET28a(+)-HNM₈₆Oct4, pET28a(+)-HNM₈₆Sox2M₈₆, pET28a(+)-HNM₈₆Klf4M₈₆, pET28a(+)-HNM₈₆cMyc 은 pET-28a(+) 벡터 (Novagen, Germany)의 다중 클로닝 부위 (multi cloning site, MCS) 중 NdeI 제한효소 부위에 본 발명에 따른 세포투과성 6 종의 역분화 전사인자 재조합 단백질로서 Nanog N-말단에 JO-86 MTD 가 융합된 재조합 단백질 HM₈₆Nanog, Oct4 N-말단에 JO-86 MTD 가 융합된 재조합 단백질 HM₈₆Oct4, Sox2 N-말단, C-말단에 JO-86 MTD 가 융합된 재조합 단백질 HM₈₆Sox2M₈₆ _, Klf4 N-말단, C-말단에 JO-86 MTD 가 융합된 재조합 단백질 HM₈₆Klf4M₈₆ _, cMyc N-말단에 JO-86 MTD 가 융합된 재조합 단백질 HM₈₆cMyc 을 코딩하는 뉴클레오티드가 삽입된 벡터를 의미한다.

본 발명의 일 실시예에서는, 단백질의 정제를 용이하게 할 목적으로 6 종의 세포투과성 역분화 전사인자 재조합 단백질의 N-말단 부위에 인위적으로 6 개의 히스티딘표지를 포함시켜 발현시키기 위하여, His-Tag 서열을 가지고 있는 pET-28a(+) 벡터 (Novagen, USA)에 본 발명의 뉴클레오티드를 클로닝한다.

상기 재조합 발현벡터로부터 발현되는 6 종의 세포투과성 역분화 전사인자 재조합 단백질의 한쪽 또는 양쪽 말단에 JO-10 MTD, JO-52 MTD, JO-84 MTD, JO-86 MTD, JO-132 MTD, JO-145 MTD, JO-173 MTD 및 JO-181 MTD 중의 하나가 융합되고, 그의 N-말단 부위에 히스티딘-표지와 NLS 가 연결되어 있다.

본 발명은 또한, 상기 재조합 발현벡터로 형질전환된 형질전환 세균을 제공한다. 본 발명의 형질전환 세균은 바람직하게는 대장균일 수 있으며, 대장균을 본 발명의 재조합 발현벡터, 예를 들면 본 발명에 따른 Nanog N-말단에 JO-86 MTD 가 융합된 세포투과성 재조합 단백질 HM₈₆Nanog 를 코딩하는 뉴클레오티드를 포함하는 재조합 발현벡터 pET28a(+)-HM₈₆Nanog 를 이용함으로써 많은 양의 세포투과성 역분화 전사인자 재조합 단백질을 발현시킬 수 있다. 형질전환은 핵산을 숙주세포에 도입하는 어떠한 방법도 포함되며, 당업자에게 공지된 형질전환 기술에 의해 수행될 수 있다. 바람직하게는 미세사출법 (microprojectile bombardment), 전기충격 유전자 전달법 (electroporation), 인산칼슘 (CaHPO₄) 침전, 염화칼슘 (CaCl₂) 침전, PEG-매개 융합법 (PEG-mediated fusion), 미세주입법 (microinjection) 및 리포좀 매개법 (liposome-mediated method) 등이 포함되나 이에 제한되지 않는다.

본 발명의 바람직한 실시예에서는, 상기의 방법으로 제조된 역분화 전사인자 6종 세포핵 배치 서열과 JO-86 MTD가 융합된 pET28a(+)-HNM₈₆Oct4를 포함하는 재조합 발현벡터를 대장균 DH5α에 형질전환시켜 수득된 형질전환 세균을 2010년 02월 17일자로 기탁번호 KCTC11640BP로 한국생명공학연구원 (Korea Research Institute of Bioscience and Biotechnology) 내 생물자원센터 (Biological Resource Center)에 기탁하고, JO-10, 181, 86, 173, 132 MTD가 융합된 pET28a(+)-HNM₁₀Nanog, pET28a(+)-HNM₁₈₁Sox2, pET28a(+)-HNM₈₆cMyc, pET28a(+)-HNM₁₇₃Klf4, pET28a(+)-HNM₁₃₂Lin28 각각을 포함하는 재조합 발현벡터를 대장균 DH5α에 형질전환시켜 수득된 형질전환 세균을 2010년 03월 10일자로 기탁번호 KCTC11660BP, KCTC11659BP, KCTC11661BP, KCTC11662BP, KCTC11663BP로 한국생명공학연구원 (Korea Research Institute of Bioscience and Biotechnology) 내 생물자원센터 (Biological Resource Center)에 기탁하였다.

본 발명은 또한, 상기 형질전환 세균을 배양하는 것을 포함하는 6 종의 세포투과성 역분화 전사인자 재조합 단백질의 제조방법을 제공한다.

상기 제조방법은 본 발명의 형질전환 세균에 도입된 재조합 발현벡터에서 본 발명의 세포투과성 역분화 전사인자 재조합 단백질을 암호화하는 폴리뉴클레오티드가 발현되도록 형질전환 세균을 적절한 배지 및 조건 하에서 배양하는 것에 의해 수행된다. 상기 형질전환 세균을 배양하여 재조합 단백질을 발현시키는 방법은 당업계에 공지되어 있으며, 예를 들면 형질전환 세균이 성장할 수 있는 적합한 배지에 접종하여 종배양한 후, 이를 본 배양용 배지에 접종하고 적합한 조건, 예컨대 유전자 발현 유도제인 아이소프로필-β-D-티오갈락토사이드 (isopropyl-β-D-thiogalactoside, IPTG)의 존재 하에서 배양함으로써 단백질의 발현을 유도할 수 있다. 배양이 완료되면, 상기 배양물로부터 실질적으로 순수한 재조합 단백질을 회수할 수 있다. 본 발명에서 용어 "실질적으로 순수한"은 본 발명의 재조합 단백질 및 이를 코딩하는 폴리뉴클레오티드의 서열이 숙주세포로부터 유래된 다른 단백질을 실질적으로 포함하지 않는 것을 의미한다.

상기 형질전환 세균에서 발현된 재조합 단백질의 회수는 당업계에 공지된 다양한 분리 및 정제 방법을 통해 수행할 수 있으며, 통상적으로 세포 조각 (cell debris), 배양 불순물 등을 제거하기 위하여 세포 용해물을 원심분리한 후, 침전, 예를 들어, 염석 (황산암모늄 침전 및 인산나트륨 침전), 용매 침전 (아세톤, 에탄올 등을 이용한 단백질 분획 침전) 등을 수행할 수 있고, 투석, 전기영동 및 각종 컬럼 크로마토그래피 등을 수행할 수 있다. 상기 크로마토그래피로는 이온교환 크로마토그래피, 겔-침투 크로마토그래피, HPLC, 역상-HPLC, 친화성 컬럼 크로마토그래피 및 한외여과 등의 기법을 단독 또는 병용하여 이용할 수 있다 (Maniatis 등, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., 1982; Sambrook 등, Molecular Cloning: A Laboratory Manual, 2d Ed., Cold Spring Harbor Laboratory Press, 1989; Deutscher, M., Guide to Protein Purification Methods Enzymology vol. 182. Academic Press. Inc., San Diego, CA,1990).

한편, 재조합 발현벡터로 형질전환된 세균에서 발현된 재조합 단백질은 단백질 분리 시 단백질의 특성에 따라 용해성 분획 (soluble fraction)과 불용성 분획 (insoluble fraction)으로 구분될 수 있다. 발현된 단백질의 대부분이 용해성 분획에 있을 경우에는 상기에 기술된 방법에 따라 별다른 어려움 없이 단백질을 분리 및 정제할 수 있다. 그러나 발현된 단백질의 대부분이 불용성 분획, 즉, 봉입체 (inclusion body) 형태로 존재하는 경우에는, 우레아, 계면활성제 등의 단백질 변성제가 포함된 용액으로 최대한 단백질을 용해시킨 후 원심분리하여 투석, 전기영동 및 각종 컬럼 크로마토그래피 등을 수행함으로써 정제할 수 있다. 이때, 단백질 변성제가 포함된 용액에 의해 단백질의 구조가 변형되어 그 활성을 잃을 수 있으므로 불용성 분획으로부터 단백질을 정제하는 과정 중에는 탈염 및 원상화 (refolding) 단계가 필요하다. 즉, 상기 탈염 및 원상화 단계는 단백질 변성제가 포함되지 않은 용액을 이용하여 투석 및 희석을 수행하거나 또는 필터를 이용하여 원심분리를 할 수 있다. 또한, 상기 용해성 분획으로부터 단백질을 정제하는 과정 중에도 정제 시 사용하는 용액 내의 염 농도가 높을 경우에는 이러한 탈염 및 원상화 단계를 수행할 수 있다.

본 발명의 일 실시양태에서는, 본 발명에 따른 세포투과성 역분화 전사인자 재조합 단백질이 봉입체 형태로서 불용성 분획에 존재함을 확인하고, 불용성 분획으로부터 이를 정제하기 위하여 용해성 분획을 트리톤 X-100 과 같은 비이온성 계면활성제를 함유하는 완충액에 용해시킨 다음 초음파로 처리하고 원심분리하여 침전물을 수득한다. 상기 수득한 침전물을 변성제인 우레아가 포함되어 있는 용액에 용해시킨 후 원심분리하여 상층액을 수득한다. 우레아를 이용하여 불용성 분획으로부터 최대한 용해된 본 발명의 재조합 단백질은 히스티딘-결합 정제 키트를 사용하여 정제하고, 이후 정제된 단백질은 투석막 등을 이용한 투석에 의해 염분의 제거 및 단백질 구조의 원상화 과정을 수행함으로써 본 발명의 재조합 단백질을 수득할 수 있다.

아울러 본 발명은 역분화 전사인자가 상실된 체세포에서의 상기 6 종의 세포투과성 역분화 전사인자 재조합 단백질을 처리함으로써 유도만능줄기세포를 확립 할 수 있는 조성물을 제공한다.

세포투과성을 갖는 역분화 전사인자를 세포의 핵내로 고효율로 도입하여 줄기세포의 특징인 만능성, 자가생산성, 미분화성을 유도하고, 줄기세포에서만 발현하는 특정 유전자들의 활성화를 통해 체세포의 역분화를 유도하여, 배아줄기세포와 유사한 유도만능줄기세포 확립시킬 수 있는 단백질로서 유용하게 사용 되어질 수 있다.

이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 자명할 것이다.

실시예 1: 재조합 단백질의 발현

상기 재조합 단백질의 유전자 컨스트럭트를 제조하기 위하여, 각각에 대해 특이적으로 고안된 프라이머 쌍과 인간 Nanog, Oct4, Sox2, Klf4,c-Myc, Lin28 cDNA 를 주형으로 사용하는 중합효소 연쇄반응 (polymerase chain reaction: PCR)을 수행하였다.

이때 HNNanog 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 187 및 188 의 염기서열을 가지고;

HNM ₈₄ Nanog 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 189 및 188 의 염기서열을 가지고;

HNNanogM ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 187 및 190 의 염기서열을 가지고;

HNM ₈₄ NanogM ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 189 및 190 의 염기서열을 가지고;

HNM ₈₆ Nanog 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 191 및 188 의 염기서열을 가지고;

HNNanogM ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 187 및 192 의 염기서열을 가지고;

HNM ₈₆ NanogM ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 191 및 192 의 염기서열을 가지고;

HNOct4 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 193 및 194 의 염기서열을 가지고;

HNM ₈₄ Oct4 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 195 및 194 의 염기서열을 가지고;

HNOct4M ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 193 및 190 의 염기서열을 가지고;

HNM ₈₄ Oct4M ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 195 및 196 의 염기서열을 가지고;

HNM ₈₆ Oct4 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 197 및 194 의 염기서열을 가지고;

HNOct4M ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 193 및 198 의 염기서열을 가지고;

HNM ₈₆ Oct4M ₈₆의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 197 및 198 의 염기서열을 가지고;

HNSox2 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 199 및 200 의 염기서열을 가지고;

HNM ₈₄ Sox2 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 201 및 200 의 염기서열을 가지고;

HNSox2M ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 199 및 202 의 염기서열을 가지고;

HNM ₈₄ Sox2M ₈₄의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 201 및 202 의 염기서열을 가지고;

HNM ₈₆ Sox2 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 203 및 200 의 염기서열을 가지고;

HNSox2M ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 199 및 204 의 염기서열을 가지고;

HNM ₈₆ Sox2M ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 203 및 204 의 염기서열을 가지고;

HNKlf4 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 205 및 206 의 염기서열을 가지고;

HNM ₈₄ Klf4 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 207 및 206 의 염기서열을 가지고;

HNKlf4M ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 205 및 208 의 염기서열을 가지고;

HNM ₈₄ Klf4M ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 207 및 208 의 염기서열을 가지고;

HNM ₈₆ Klf4 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 209 및 206 의 염기서열을 가지고;

HNKlf4M ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 205 및 210 의 염기서열을 가지고;

HNM ₈₆ Klf4M ₈₆의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 209 및 210 의 염기서열을 가지고;

HNcMyc 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 211 및 212 의 염기서열을 가지고;

HNM ₈₄ cMyc 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 213 및 212 의 염기서열을 가지고;

HNcMycM ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 211 및 214 의 염기서열을 가지고;

HNM ₈₄ cMycM ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 213 및 214 의 염기서열을 가지고;

HNM ₈₆ cMyc 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 215 및 212 의 염기서열을 가지고;

HNcMycM ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 211 및 216 의 염기서열을 가지고;

HNM ₈₆ cMycM ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 215 및 216 의 염기서열을 가지고;

HNLin28 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 217 및 218 의 염기서열을 가지고;

HNM ₈₄ Lin28 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 219 및 218 의 염기서열을 가지고;

HNLin28M ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 217 및 220 의 염기서열을 가지고;

HNM ₈₄ Lin28M ₈₄ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 219 및 220 의 염기서열을 가지고;

HNM ₈₆ Lin28 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 221 및 218 의 염기서열을 가지고;

HNLin28M ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 217 및 222 의 염기서열을 가지고;

HNM ₈₆ Lin28M ₈₆ 의 증폭을 위한 정방향 및 역방향 프라이머는 각각 서열번호: 221 및 222 의 염기서열을 갖는다.

PCR 반응은 주형 유전자로 인간 Nanog, Oct4, Sox2, Klf4,c-Myc, Lin28 cDNA 각 100 ng, 각 0.2 mM의 최종 농도 dNTP 혼합물 (dGTP, dATP, dTTP 및 dCTP, 각 2 mM), 0.5 μM 의 각 프라이머, 10× Taq 완충용액 10 ㎕, Taq 중합효소 (Takara, Japan) 0.5 ㎕를 포함하는 용액을 최종 부피 100 ㎕ 반응액으로 하여 수행하였다. PCR 반응 조건은 먼저 95℃에서 2 분간 열 변성 (denaturing)시킨 후 95℃에서 45 초, 67℃에서 45 초 및 72℃에서 45 초의 반응을 30 회 반복하였고, 최종적으로 72℃에서 5 분간 증폭하였다. 반응이 끝난 후 0.8％ 아가로즈 겔에 전기영동 (electrophoresis)을 수행하여 증폭된 생성물을 확인하였다.

아가로즈 겔에서 증폭된 재조합 단편을 회수한 후 이들 각각을 상용의 키트 (QIAquick Gel extraction kit; Qiagen, USA)를 이용하여 추출, 정제하였다. 추출한 단편을 pGEM-T Easy 벡터 (Promega, USA)에 삽입한 후 MTD 가 융합된 Nanog, Oct4, Sox2, Klf4,c-Myc, Lin28 재조합 단백질 유전자 단편이 삽입된 pGEM-T Easy 벡터를 대장균 DH5α 감응세포 (competent cell)에 형질전환시켰다. 이를 50 ㎍/㎖ 암피실린 (ampicillin)이 포함된 평판 LB 배지에 접종하고 37℃에서 밤새 배양하여 형질전환된 대장균을 선별하고, 이를 다시 액체 LB 배지에서 배양한 후, 이로부터 각각의 Nanog, Oct4, Sox2, Klf4,c-Myc, Lin28 재조합 단백질 유전자가 삽입된 pGEM-T Easy 벡터를 다량으로 수득하였다.

pGEM-T Easy 벡터에 삽입된 재조합 단편을 NdeI 제한효소 (Enzynomics, Korea)로 분리하여 0.8％ 아가로스 겔에 전기 영동한 것으로부터 각각의 재조합 단편이 상기 벡터에 올바르게 삽입되었음을 확인하였다.

상기에서 각각의 Nanog, Oct4, Sox2, Klf4,c-Myc, Lin28 재조합 단백질 유전자가 삽입된 pGEM-T Easy 벡터를 제한효소 NdeI 을 사용해 37℃에서 2 시간 동안 절단 (digestion)하여 각각의 재조합 단편을 얻었다. 아가로즈 겔에서 재조합 단편을 회수한 후 이들 각각을 상용의 키트 (QIAquick Gel extraction kit; Qiagen, USA)를 이용하여 추출, 정제하였다. 한편, 히스티딘-표지 (histidine-tag)와 T7 프로모터를 가진 발현벡터 pET-28a(+) 벡터 (Novagen, USA)를 제한효소 NdeI 을 사용해 상기와 동일한 조건으로 절단하였다. 상기에서 추출된 각각의 재조합 단편과 절단된 pET-28a(+) 벡터를 혼합하고, 여기에 T4 DNA 연결효소 (ligase; Takara, 일본)를 첨가한 후 16℃에서 12 시간 동안 라이게이션 (ligation)시킨 다음, 대장균 DH5α 감응세포에 형질전환시켜 재조합 단백질 발현벡터를 수득하였다,

pET-28a(+) 벡터에 삽입된 재조합 단편을 NdeI 제한효소로 분리하여 0.8％ 아가로스 겔에 전기영동한 것으로부터 각각의 재조합 단편이 상기 벡터에 올바르게 삽입되었음을 확인하였다.

이렇게 수득된 재조합 단백질 발현벡터를 각각 pET28a(+)-HNNanog, pET28a(+)-HNM₈₄Nanog, pET28a(+)-HNNanogM₈₄, pET28a(+)-HNM₈₄NanogM₈₄, pET28a(+)-HNM₈₆Nanog, pET28a(+)-HNNanogM₈₆, pET28a(+)-HNM₈₆NanogM₈₆, pET28a(+)-HNOct4, pET28a(+)-HNM₈₄Oct4, pET28a(+)-HNOct4M₈₄, pET28a(+)-HNM₈₄Oct4M₈₄, pET28a(+)-HNM₈₆Oct4, pET28a(+)-HNOct4M₈₆, pET28a(+)-HNM₈₆Oct4M₈₆, pET28a(+)-HNSox2, pET28a(+)-HNM₈₄Sox2, pET28a(+)-HNSox2M₈₄, pET28a(+)-HNM₈₄Sox2M₈₄, pET28a(+)-HNM₈₆Sox2, pET28a(+)-HNSox2M₈₆, pET28a(+)-HNM₈₆Sox2M₈₆, pET28a(+)-HNKlf4, pET28a(+)-HNM₈₄Klf4, pET28a(+)-HNKlf4M₈₄, pET28a(+)-HNM₈₄Klf4M₈₄, pET28a(+)-HNM₈₆Klf4, pET28a(+)-HNKlf4M₈₆, pET28a(+)-HNM₈₆Klf4M₈₆, pET28a(+)-HNcMyc, pET28a(+)-HNM₈₄cMyc, pET28a(+)-HNcMycM₈₄, pET28a(+)-HNM₈₄cMycM₈₄, pET28a(+)-HNM₈₆cMyc, pET28a(+)-HNcMycM₈₆, pET28a(+)-HNM₈₆cMycM₈₆ , pET28a(+)-HNLin28, pET28a(+)-HNM₈₄Lin28, pET28a(+)-HNLin28M₈₄, pET28a(+)-HNM₈₄Lin28M₈₄, pET28a(+)-HNM₈₆Lin28, pET28a(+)-HNLin28M₈₆, pET28a(+)-HNM₈₆Lin28M₈₆ 으로 명명하였다.

대장균 BL21 CodonPlus (DE3)에 재조합 발현벡터 pET28a(+)-HNNanog, pET28a(+)-HNM₈₄Nanog, pET28a(+)-HNNanogM₈₄, pET28a(+)-HNM₈₄NanogM₈₄, pET28a(+)-HNM₈₆Nanog, pET28a(+)-HNNanogM₈₆, pET28a(+)-HNM₈₆NanogM₈₆, pET28a(+)-HNM₁₀Nanog, pET28a(+)-HNOct4, pET28a(+)-HNM₈₄Oct4, pET28a(+)-HNOct4M₈₄, pET28a(+)-HNM₈₄Oct4M₈₄, pET28a(+)-HNM₈₆Oct4, pET28a(+)-HNOct4M₈₆, pET28a(+)-HNM₈₆Oct4M₈₆, pET28a(+)-HNM₅₂Oct4, pET28a(+)-HNSox2, pET28a(+)-HNM₈₄Sox2, pET28a(+)-HNSox2M₈₄, pET28a(+)-HNM₈₄Sox2M₈₄, pET28a(+)-HNM₈₆Sox2, pET28a(+)-HNSox2M₈₆, pET28a(+)-HNM₈₆Sox2M₈₆, pET28a(+)-HNM₁₈₁Sox2, pET28a(+)-HNKlf4, pET28a(+)-HNM₈₄Klf4, pET28a(+)-HNKlf4M₈₄, pET28a(+)-HNM₈₄Klf4M₈₄, pET28a(+)-HNM₈₆Klf4, pET28a(+)-HNKlf4M₈₆, pET28a(+)-HNM₈₆Klf4M₈₆, pET28a(+)-HNM₁₇₃Klf4, pET28a(+)-HNcMyc, pET28a(+)-HNM₈₄cMyc, pET28a(+)-HNcMycM₈₄, pET28a(+)-HNM₈₄cMycM₈₄, pET28a(+)-HNM₈₆cMyc, pET28a(+)-HNcMycM₈₆, pET28a(+)-HNM₈₆cMycM₈₆ , pET28a(+)-HNM₁₄₅cMyc, pET28a(+)-HNLin28, pET28a(+)-HNM₈₄Lin28, pET28a(+)-HNLin28M₈₄, pET28a(+)-HNM₈₄Lin28M₈₄, pET28a(+)-HNM₈₆Lin28, pET28a(+)-HN Lin28M₈₆, pET28a(+)-HNM₈₆Lin28M₈₆, pET28a(+)-HNM₁₃₂Lin28 각각을 열 충격 방법으로 형질전환 시킨 후 50 ㎍/㎖의 카나마이신이 함유된 LB 배지에서 배양하였다. 이후 상기 재조합 단백질 유전자가 도입된 대장균을 25 ㎖ LB 배지에 접종하고 37℃에서 밤새도록 배양한 후, 이를 다시 1 ℓ LB 배지에 접종하고 37℃에서 OD₆₀₀이 0.6 내지 0.7에 도달할 때까지 배양하였다. 이 배양액에 단백질 발현의 유도제로서 0.65 mM의 IPTG를 첨가하고 37℃에서 3시간 동안 추가로 배양하였다. 이 배양액을 4℃에서 4,000 ×g, 20분간 원심분리하여 상등액을 제거하고 균체를 회수하였다. 회수한 균체를 용해 완충액 (100 mM NaH₂PO₄, 10 mM Tris-HCl, 8 M 우레아, pH 8.0)에 현탁한 후 초음파 처리를 수행하여 세포를 파쇄하였으며, 이를 14,000 ×g로 15분간 원심분리하여 용해성 분획과 불용성 분획을 얻었다. 이 용해성 분획과 불용성 분획을 각각 SDS-PAGE에 걸어 단백질 발현 특성과 발현량 정도를 분석하였다.

도 2 에 나타난 바와 같이, 약 34 kDa 의 Nanog, 약 40 kDa 의 Oct4, 약 35 kDa 의 Sox2, 약 52 kDa 의 Klf4 와 약 50 kDa 의 cMyc 은 본 발명의 세포투과성 전사인자 재조합 단백질은 봉입체 형태로 대부분 불용성 분획에 포함되어 있으며, IPTG 무처리 배양액 (-)에 비해 IPTG 처리 배양액(+)에서 목적 단백질의 발현이 현저히 증가되었음을 확인하였다.

실시예 2: 재조합 단백질의 정제

본 발명에 따른 세포투과성 전사인자 재조합 단백질은 봉입체 형태로 불용성 분획에 존재하므로 이를 정제하기 위하여 강력한 변성제로서 8 M 우레아 (urea)를 이용하였다.

먼저, 본 발명의 재조합 발현벡터 각각으로 형질전환된 BL21 CodonPlus (DE3) 균주를 상기 실시예 1 과 같이 1 ℓ의 LB 배지에 배양하였다. 각각의 배양액을 원심분리하여 수득한 균체를 20 ㎖의 용해 완충액 (100 mM NaH₂PO₄, 10 mM Tris-HCl, 8 M 우레아, pH 8.0)에 기포가 생기지 않도록 주의하면서 현탁하고, 이를 마이크로팁 (microtip)이 장착된 초음파 균질기를 이용하여 저온에서 균체를 파쇄하였다. 이때, 처리시간은 장치의 출력을 최대출력의 25％로 설정하고, 7 분간 45 초 처리 후 10 초 방치를 반복하였다. 충분히 용균된 봉입체를 4℃에서 4,000 ×g, 20 분간 원심분리하여 침전물을 제거하고 상층액을 회수하였다. 회수된 상층액을 나이트릴로트라이아세트산 아가로스 (nitrilotriacetic acid agarose)에 Ni 를 부여한 Ni-NTA 아가로스 레진에 로우딩하였다. 이때 Ni-NTA 아가로스는 미리 용해 완충액으로 수차례 세척하여 평형화시킨 후 사용하였다. 상층액을 4℃에서 8 시간 이상 진탕기로 천천히 교반하면서 레진에 흡착시켰다. 재조합 단백질이 포함된 봉입체가 흡착된 레진을 4℃의 분당 회전수 1,000 rpm 에서 5 분간 원심분리하여 반응액을 제거하였고, 비특이적 흡착 물질을 제거하기 위해 레진을 세척 완충액 (washing buffer: 100 mM NaH₂PO₄, 10 mM Tris-HCl, 8 M 우레아, pH 6.3)을 이용하여 5 회 세척하였다. 세척된 레진에 pH 4.0 의 산성 조건에서 레진 용적 2 배의 용출 완충액 (elution buffer: 100 mM NaH₂PO₄, 10 mM Tris-HCl, 8 M 우레아, pH 4.5)을 로우딩하고 진탕기에서 2 시간, 또는 8 시간 이상을 교반하여 단백질을 용출하였다. 용출된 단백질의 순도를 검정하기 위하여 12％ SDS-PAGE 겔에서 전기영동을 수행한 후 겔을 쿠마시 브릴리언트 블루 R (coomassie brilliant blue R)로 가볍게 진탕하면서 염색하고, 목적 단백질의 밴드가 명확해질 때까지 탈색액을 이용하여 탈색하였다.

그 결과, 도2에 나타난 바와 같이, 마커 단백질의 전기영동 위치와 비교하여 JO-84 MTD 및 JO-86 MTD가 각각 융합된 모든 세포투과성 전사인자 재조합 단백질이 각각의 부위에서 검출되어 불용성 분획으로부터 정제되었음을 확인하였다.

단백질의 활성 재생을 위해서 불용성 분획으로부터 정제된 본 발명의 재조합 단백질은 강력한 변성제인 8 M 우레아에 의해 변성되었기 때문에 이를 활성 형태로 전환시키기 위하여 하기와 같이 원상화 과정을 수행하였다.

먼저, 정제된 재조합 단백질을 원상화 완충액 (refolding buffer: 0.55 M 구아니딘 [Guanidine] HCl, 0.88 M L-아르기닌, 50 mM 트리스-HCl, 150 mM NaCl, 1 mM EDTA, 100 mM NDSB, 1 mM 산화형 글루타치온 [glutathione oxidized] 및 1 mM 환원형 글루타치온 [glutathione reduced])을 이용하여 4℃에서 72 시간 이상 투석하여 변성제를 제거함으로써 재조합 단백질이 재활성, 즉 원상화되도록 하였다. 이때 24 시간 마다 용기 내 원상화 완충액을 교환해주었다. 이후 활성화된 재조합 단백질을 세포 배양용 배지인 1％ 페니실린/스트렙토마이신이 첨가된 DMEM (Dulbecco's Modified Eagle Medium)에 대해 투석막 (Snakeskin pleated, PIERCE)을 사용하여 4℃에서 9 시간 동안 교반하면서 투석하였다. 이때 3 시간 마다 용기 내 DMEM 을 교환해주었다. 상기 원상화 과정에 의해 활성 형태로 전환된 세포투과성 전사인자 재조합 단백질을 이후의 실험에 사용하였다.

실시예 3: 역분화 줄기세포 유도 및 알칼라인 포스파타아제 염색 ( AP Staining )

인간 피부 섬유아세포 (human dermal fibroblast, HDF)를 배양하여 역분화 줄기세포를 유도하는 실험을 실시하였다. 처음 7 일 동안은 HDF 전용 배양액 (M106 (Cascade Biologic™) + LSGS (Cascade Biologic™)) 에서 37℃, 5％ CO₂의 습윤 조건 하에서 배양하였다. 그 이후 14 일 간은 인간 줄기세포 전용 배양액 (DMEM/F12 (HyClone？), 20％ KSR (GIBCO？), 2mM L-glutamin (GIBCO？), 2mM MEM Non Essential Amino Acid (GIBCO？), 0.1mM β-mercaptoethanol (GIBCO？), 0.1％ penicillin/streptomycin (GIBCO？))에서 배양하였다. 5 종의 세포 투과성 역분화 전사인자 재조합 단백질과 이에 대한 대조군으로 MTD 를 융합하지 않은 역분화 전사인자 재조합 단백질을 각 1μM 씩 처리하여 주고 이는 16 일간 매일매일 배양액과 단백질을 바꿔준다. 단백질 처리 16 일 후에 형성된 콜로니는 총 23 개이며 이렇게 형성된 콜로니는 역분화 줄기세포임을 증명하는 실험에 사용되었다.

역분화 줄기세포 콜로니임을 확인 하기 위해 알칼라인 포스파타아제 염색 (Sigma)을 시도하였다. 배아줄기세포는 알칼라인 포스파타아제의 발현 유무에 따라 미분화/분화의 특징으로 구분된다. 알칼라인 포스파타아제는 인체의 여러 장기에도 분포하지만 배아줄기세포의 세포질에 존재하며 이는 염색 후 붉은색으로 나타나는 것으로써 확인된다. 알칼라인 포스파타아제 염색 키트에 포함되어 있는 Fast Blue RR Salt capsule을 물에 용해한다.

Naphthol AS-MX 알칼리성 인산 용액을 희석된 디아조늄 염용액과 섞어준다. 고정용액으로 6-well plate안의 세포들을 약 30초간 고정 해준 후 물로 부드럽게 45초 동안 세척하여 준다. 그 후 알칼라인 염색용액을 추가한 후 실온에서 30분간 배양한다. 이때 빛에 노출되지 않게 주의하고 배양시간이 끝난 후 물로 세척하여 준다. Mayer' s Hematoxylin용액으로 10분간 대조염색을 한 후 2분간 물로 세척을 하여 준다. 그 후 mounting 용액으로 세포를 덮어 준 후 현미경으로 색깔의 변화를 관찰 한다.

그 결과 도 3 과 같이 마우스 배아줄기세포의 알칼라인 포스파타아제 염색이 되어 배아줄기세포의 특징인 미분화능이 확인된다. 세포 투과성 역분화 전사인자 재조합 단백질을 16 일 동안 처리하여 생성된 콜로니에서도 미분화 마커인 알칼라인 포스파타아제로 붉은색 염색이 되는 것을 확인하였다.

실시예 4: 면역염색법 ( immunocyto chemistry )

상기 실시예 3 에 명시한 바와 같이 세포투과성 전사인자 재조합 단백질을 인간 피부 섬유아세포 (human dermal fibroblast, HDF)에 처리하여 얻은 유도만능줄기세포 (induced pluripotent stem cell) 콜로니 (colony)를 얻었다. 모세유리관을 알코올램프에 가열 후 구부려 얻은 고리모양 유리관을 이용하여 콜로니를 떼내었다. 콜로니가 제대로 떨어진 것을 실체현미경 (stereomicroscope)을 통해 확인하였으며 이를 인간 피부 섬유아세포가 배양되어 있는 8-well 챔버 슬라이드 (chamber slide, NUNC) 이 생착할 수 있게 얹어놓았다. 하루 동안의 안정화를 거친 뒤, 챔버 슬라이드의 배지를 제거하고 PBS (phosphate buffered saline, WelGENE)로 2 회 세척하였다. 2％ 파라포름알데하이드 (paraformaldehyde, JUNSEI)를 20 분간 처리하여 콜로니를 고정하였다. 다시 PBS 로 2 회 세척 후, 0.1％ 트라이톤-X100 을 5 분간 처리하여 세포막의 침투성을 부여하였다. PBS 로 2 회 세척 후, 2％ BSA (bovine serum albumin, sigma)/PBS 를 상온에서 1 시간 동안 처리하여 비특이적인 단백질의 방해를 막았다. Nanog 에 대한 1 차 항체로 abcam 사의 Nanog 항체 (anti-Nanog antibody, abcam)를, Oct4 에 대한 1 차 항체로서 abcam 사의 Oct4 항체 (anti-Oct4 antibody, abcam)를 2％ BSA/PBS 에 1:1000 희석하여 콜로니에 처리하였다. 4℃ 냉장고에서 밤새 반응 시킨 뒤, PBS 로 2 회 세척 후 2 차 항체를 상온에서 1 시간 처리하였다. Nanog 항체에 대하여 항 rabbit IgG 항체 Alexa Fluor^？ 488 (Goat anti-rabbit IgG, Alexa)를, Oct4 항체에 대하여 항 goat IgG 항체 Alexa Fluor^？ 546 (Rabbit anti-goat IgG, Alexa)를 2％ BSA/PBS 에 1:500 희석하여 사용하였다. PBS 로 3 번 세척 후 DAPI(4,6-Diamidino-2-phenylindole, sigma) 300nM 로 빛을 차단하여 상온에서 5 분간 핵염색 하였다. PBS 로 3 회 세척한 뒤, 중첩 배지 (mounting medium. VECTOR)을 슬라이드 위에 점적한 후 15 분 후에 동초점 레이저 주사 현미경으로 관찰하였다. 이때 동초점 레이저 주사 현미경은 노마르스키 필터 (Nomarski filter)를 이용하여 세포의 원형, Nanog 는 488nm 에서 Oct4 는 546nm 에서 여기되는 형광을 관찰하였다.

그 결과, 도 4 에 나타난 바와 같이, 본 발명에 따른 세포투과성 전사인자 재조합 단백질로 인한 유도만능줄기세포 콜로니가 줄기세포의 특징으로 간주되는 Nanog 와 Oct4 를 발현하는 것을 확인하였다.

실시예 5: 역분화 전사인자로 확립된 유도만능줄기세포의 세포표면의 줄기세포 특이적마커 발현

상기 실시예 에서 AP staining 으로 줄기세포의 미분화를 확인된 colony 를 트립신/EDTA (T/E, Invitrogen)으로 37℃에서 3 분 처리하여 하나의 세포로 분리 한다. 0.1％ 소혈청알부민 (Bovine Serum Albumin)이 첨가된 PBS (phosphate buffered saline)로 2 회 세척하였다. Fc 부분이 cell surface 의 FcR 에 non-specific binding 하는 것을 block 하기 위해서 Fc Blocker CD16/32 (Fc Blocker, BD Pharmingen)로 10min 처리하고 0.1％ 소혈청알부민(Bovine Serum Albumin)이 첨가된 PBS (phosphate buffered saline)로 2 회 세척하였다. FITC (fluorescein-5-isothiocyanate)가 표지된 SSEA-4 (Stage Specific Embryonic Antigen-4) antibody (BD bioscience), PE (Phycoerythrin)가 표지된 SSEA-3 (Stage Specific Embryonic Antigen-3) antibody (BD bioscience)를 TRA-1-60 (Tumor Rejection Antigen, BD bioscience) 1μg/1x10⁶ 농도로 얼음에서 20 분 염색하였다. 0.1％ 소혈청알부민 (Bovine Serum Albumin)이 첨가된 PBS (phosphate buffered saline, Welgene)로 2 회 세척한 후, 준비된 세포에 대해 셀퀘스트 프로 (CellQuest Pro, BD)세포측정 분석 소프트웨어 (CellQuest Pro cytometric analysis software)를 이용한 Calibur 유세포 분석기 (flow cytometry, Becton Dickinson, CA)로 분석하였다.

그 결과, 도 5 에 나타난 바와 같이, 본 발명에 따른 세포투과성 역분화 전사인자 재조합 단백질로 유도 되어진 colony 의 줄기세포 특이적 세포표면 마커 (stem cell specific surface marker) 발현정도는 대조군에 비하여 높은 형광 intensity 를 나타냄을 확인하였다.

도 5 에서 no staining 은 역분화 전사인자를 처리하지 않은 그룹이며 SSEA-3 를 발현하는 세포는 0.5％이다. MTD 융합되지 않은 대조군은 4.5％인 반면 세포투과성 역분화 전사인자를 처리하여 형성된 colony 에서는 9.9％의 세포가 SSEA-3 를 발현하였다.

그 결과, 도 5 에 나타난 바와 같이, 본 발명에 따른 세포투과성 역분화 전사인자 재조합 단백질로 유도 되어진 colony 의 줄기세포 특이적 세포표면 마커 (stem cell specific surface marker), TRA-1-60 의 발현정도가 5％이상 대조군에 비해 높은것을 확인하였다.

실시예 6: 역전사 중합효소 연쇄반응( RT - PCR )

상기 실시예에서와 같은 방법으로 역분화 줄기세포를 유도하는 과정에서 줄기세포 마커 유전자의 발현이 증가함을 확인하기 위하여 역전사 중합효소 연쇄 반응 실험을 실험 하였다. 트라이졸 용액 (iNTRON)으로 세포의 RNA를 추출 하였고 이 중 5μg에 해당하는 양을 정량하여 역전사 (Roche) 시킨다. 단백질을 처리하지 않은 대조군과 세포투과성 역분화 전사인자 재조합 단백질을 처리 하여 역분화 줄기세포로 유도 한 군의 cDNA 에서 줄기세포 마커 유전자의 발현차이를 확인하기 위하여 중합효소 연쇄 반응 (PCR)을 수행 하였다. 이때 확인한 줄기세포 마커 유전자는 Oct4, Nanog, LEFTB. Klf4, cMyc, EBAF, UTF1, CD9, ESG1, IFITM1, GAL, BRIX, REX1이며 PCR 대조군으로써 모든 세포에서 고루 발현되는 유전자인 GAPDH를 선택하여 총 14개의 유전자에 대한 PCR을 수행하였다. 이는 95℃에서 1분, 55℃에서 1분, 72℃에서 1분씩 반응 하며 이것을 30~35회 반복한다. 반응이 끝난 PCR 산물을 아가로즈 젤에 로딩 하여 전기영동을 수행하였다.

그 결과 도 6 에 나타난 바와 같이 세포 투과성 역분화 전사인자 재조합단백질을 처리 할 경우 아무런 단백질을 처리하지 않은 대조군에 줄기세포에서 특이적으로 발현하는 유전자가 증가하는 것을 확인하였다.

실시예 7: 역분화 줄기세포 유도

인간 피부 섬유아세포 (human dermal fibroblast, HDF)를 배양하여 역분화 줄기세포를 유도하는 실험을 실시하였다. 도 7 에 나타난 바와 같이 총 23 일 동안 인간 줄기세포 전용 배양액 (DMEM/F12 (HyClone？), 20％ KSR (GIBCO？), 2mM L-glutamin (GIBCO？), 2mM MEM Non Essential Amino Acid (GIBCO？), 0.1mM β-mercaptoethanol (GIBCO？), 0.1％ penicillin/streptomycin (GIBCO？))에서 배양하였다. 5 종의 세포 투과성 역분화 전사인자 재조합 단백질과 이에 대한 대조군으로 MTD 를 융합하지 않은 역분화 전사인자 재조합 단백질을 각 8 μg/ml 혹은 20 μg/ml 씩 처리하여 주고 이는 10 일간 매일매일 배양액과 단백질을 바꿔준다. 단백질 처리 8 일 후에 형성된 콜로니는 총 13 개이며 이렇게 형성된 콜로니는 단백질 처리 10 일 후에 모세유리관을 알코올램프에 가열 후 구부려 얻은 고리모양 유리관을 이용하여 떼내었다. 콜로니가 제대로 떨어진 것을 실체현미경 (stereomicroscope)을 통해 확인하였으며 이후 단백질을 더 이상 처리하지 않는 군과 콜로니를 떼어낸 8 일 이후 다시 5 일 동안 단백질을 16 μg/ml 씩 처리하는 군으로 나누었다. 이 콜로니들의 모습은 도 7 에 나타난 바와 같으며 이 후 역분화 줄기세포임을 증명하는 실험에 사용되었다.

실시예 8: 면역염색법 ( immunocyto chemistry )

상기 실시예 7 에 명시한 바와 같이 세포투과성 전사인자 재조합 단백질을 인간 피부 섬유아세포 (human dermal fibroblast, HDF)에 처리하여 얻은 유도만능줄기세포 (induced pluripotent stem cell) 콜로니 (colony)를 쥐의 피부 섬유아세포가 배양되어 있는 8-well 챔버 슬라이드 (chamber slide, NUNC) 이 생착할 수 있게 얹어놓았다. 하루 동안의 안정화를 거친 뒤, 챔버 슬라이드의 배지를 제거하고 PBS (phosphate buffered saline, WelGENE)로 2 회 세척하였다. 2％ 파라포름알데하이드 (paraformaldehyde, JUNSEI)를 20 분간 처리하여 콜로니를 고정하였다. 다시 PBS 로 2 회 세척 후, 0.1％ 트라이톤-X100 을 5 분간 처리하여 세포막의 침투성을 부여하였다. PBS 로 2 회 세척 후, 2％ BSA (bovine serum albumin, sigma)/PBS 를 상온에서 1 시간 동안 처리하여 비특이적인 단백질의 방해를 막았다. Nanog 에 대한 1 차 항체로 abcam 사의 Nanog 항체 (anti-Nanog antibody, abcam)를, Oct4 에 대한 1 차 항체로서 abcam 사의 Oct4 항체 (anti-Oct4 antibody, abcam)를 2％ BSA/PBS 에 1:1000 희석하여 콜로니에 처리하였다. 4℃ 냉장고에서 밤새 반응 시킨 뒤, PBS 로 2 회 세척 후 2 차 항체를 상온에서 1 시간 처리하였다. Nanog 항체에 대하여 항 rabbit IgG 항체 Alexa Fluor^？ 488 (Goat anti-rabbit IgG, Alexa)를, Oct4 항체에 대하여 항 goat IgG 항체 Alexa Fluor^？ 546 (Rabbit anti-goat IgG, Alexa)를 2％ BSA/PBS 에 1:500 희석하여 사용하였다. PBS 로 3 번 세척 후 PI 300nM 로 빛을 차단하여 상온에서 5 분간 핵염색 하였다. PBS 로 3 회 세척한 뒤, 중첩 배지 (mounting medium. VECTOR)을 슬라이드 위에 점적한 후 15 분 후에 동초점 레이저 주사 현미경으로 관찰하였다. 이때 동초점 레이저 주사 현미경은 노마르스키 필터 (Nomarski filter)를 이용하여 세포의 원형, Nanog 는 488nm 에서 Oct4 는 546nm 에서 여기되는 형광을 관찰하였다.

그 결과, 도 8 에 나타난 바와 같이, 본 발명에 따른 세포투과성 전사인자 재조합 단백질로 인한 유도만능줄기세포 콜로니가 줄기세포의 특징으로 간주되는 Nanog 와 Oct4 를 발현하는 것을 확인하였다.

실시예 9: 역분화 전사인자로 확립된 유도만능줄기세포의 세포표면의 줄기세포 특이적마커 발현

상기 실시예 에서 AP staining 으로 줄기세포의 미분화를 확인된 colony 를 트립신/EDTA (T/E, Invitrogen) 으로 37℃에서 3 분 처리하여 하나의 세포로 분리 한다. 0.1％ 소혈청알부민(Bovine Serum Albumin)이 첨가된 PBS (phosphate buffered saline)로 2 회 세척하였다. Fc 부분이 cell surface 의 FcR 에 non-specific binding 하는것을 block 하기 위해서 Fc Blocker CD16/32 (Fc Blocker, BD Pharmingen)로 10min 처리하고 0.1％ 소혈청알부민(Bovine Serum Albumin)이 첨가된 PBS (phosphate buffered saline)로 2 회 세척하였다. FITC (fluorescein-5-isothiocyanate)가 표지된 SSEA-4 (Stage Specific Embryonic Antigen-4) antibody (BD bioscience), PE (Phycoerythrin)가 표지된 SSEA-3 (Stage Specific Embryonic Antigen-3) antibody (BD bioscience)를 TRA-1-60 (Tumor Rejection Antigen, BD bioscience) 1μg/1x10⁶ 농도로 얼음에서 20 분 염색하였다. 0.1％ 소혈청알부민(Bovine Serum Albumin)이 첨가된 PBS (phosphate buffered saline, Welgene)로 2 회 세척한 후, 준비된 세포에 대해 셀퀘스트 프로 (CellQuest Pro, BD)세포측정 분석 소프트웨어 (CellQuest Pro cytometric analysis software)를 이용한 Calibur 유세포 분석기 (flow cytometry, Becton Dickinson, CA)로 분석하였다.

그 결과, 도 9 에 나타난 바와 같이, 본 발명에 따른 세포투과성 역분화 전사인자 재조합 단백질로 유도 되어진 colony 의 줄기세포 특이적 세포표면 마커 (stem cell specific surface marker) 발현정도는 대조군에 비하여 높은 형광 intensity 를 나타냄을 확인하였다.

도 9 에서 HDF 는 역분화 전사인자를 처리하지 않은 대조군이며 SSEA-3 를 발현하는 세포는 약 1％ 미만이다. 세포투과성 역분화 전사인자를 처리하여 형성된 colony 에서는 각각 약 5％와 약 30％의 세포가 SSEA-3 를 발현하였다.

그 결과, 도 9 에 나타난 바와 같이, 본 발명에 따른 세포투과성 역분화 전사인자 재조합 단백질로 유도 되어진 colony 의 줄기세포 특이적 세포표면 마커 (stem cell specific surface marker), TRA-160 의 발현정도가 약 10~12％이상 대조군에 비해 높은것을 확인하였다.

실시예 10: Codon optimization 된 재조합 단백질의 발현

Genscript사에서 합성한 codon optimized 된 재조합 단백질에 대한 발현 벡터 pET28a(+)-HNM₁₀Nanog, pET28a(+)-HNM₅₂Oct4, pET28a(+)-HNM₁₈₁Sox2, pET28a(+)-HNM₁₇₃Klf4, pET28a(+)-HNM₁₄₅cMyc, pET28a(+)-HNM₁₃₂Lin28를 대장균 BL21 CodonPlus (DE3)에 각각을 열 충격 방법으로 형질전환 시킨 후 50 ㎍/㎖의 카나마이신이 함유된 LB 배지에서 배양하였다. 이후 상기 재조합 단백질 유전자가 도입된 대장균을 25 ㎖ LB 배지에 접종하고 37℃에서 밤새도록 배양한 후, 이를 다시 1 ℓ LB 배지에 접종하고 37℃에서 OD₆₀₀이 0.6 내지 0.7에 도달할 때까지 배양하였다. 이 배양액에 단백질 발현의 유도제로서 0.65 mM의 IPTG를 첨가하고 37℃에서 3시간 동안 추가로 배양하였다. 이 배양액을 4℃에서 4,000 ×g, 20분간 원심분리하여 상등액을 제거하고 균체를 회수하였다. 회수한 균체를 용해 완충액 (100 mM NaH₂PO₄, 10 mM Tris-HCl, 8 M 우레아, pH 8.0)에 현탁한 후 초음파 처리를 수행하여 세포를 파쇄하였으며, 이를 14,000 ×g로 15분간 원심분리하여 용해성 분획과 불용성 분획을 얻었다. 이 용해성 분획과 불용성 분획을 각각 SDS-PAGE에 걸어 단백질 발현 특성과 발현량 정도를 분석하였다.

그 결과, 도 10 과 같은 구조를 갖는 재조합 단백질 유전자의 발현 벡터는 도 11 에 나타난 바와 같이, 약 34 kDa 의 Nanog, 약 40 kDa 의 Oct4, 약 35 kDa 의 Sox2, 약 52 kDa 의 Klf4 와 약 50 kDa 의 cMyc 은 본 발명의 세포투과성 전사인자 재조합 단백질은 봉입체 형태로 대부분 불용성 분획에 포함되어 있으며, IPTG 무처리 배양액 (-)에 비해 IPTG 처리 배양액(+)에서 목적 단백질의 발현이 codon optimization 되기 전보다 현저히 증가되었음을 확인하였다.

한국생명공학연구원

KCTC11640BP

20100217

<110> Procell Therapeutics Inc. JO, Daewoong <120> Establishment of Induced Pluripotent Stem (iPS) Cell Line With Reprogramming CP-TFs for Patient-Specific Stem Cell Therapy <130> FC103121 <150> US 61/159,461 <151> 2009-03-12 <160> 252 <170> KopatentIn 1.71 <210> 1 <211> 918 <212> DNA <213> Human <400> 1 atgagtgtgg atccagcttg tccccaaagc ttgccttgct ttgaagcatc cgactgtaaa 60 gaatcttcac ctatgcctgt gatttgtggg cctgaagaaa actatccatc cttgcaaatg 120 tcttctgctg agatgcctca cacggagact gtctctcctc ttccttcctc catggatctg 180 cttattcagg acagccctga ttcttccacc agtcccaaag gcaaacaacc cacttctgca 240 gagaagagtg tcgcaaaaaa ggaagacaag gtcccggtca agaaacagaa gaccagaact 300 gtgttctctt ccacccagct gtgtgtactc aatgatagat ttcagagaca gaaatacctc 360 agcctccagc agatgcaaga actctccaac atcctgaacc tcagctacaa acaggtgaag 420 acctggttcc agaaccagag aatgaaatct aagaggtggc agaaaaacaa ctggccgaag 480 aatagcaatg gtgtgacgca gaaggcctca gcacctacct accccagcct ttactcttcc 540 taccaccagg gatgcctggt gaacccgact gggaaccttc caatgtggag caaccagacc 600 tggaacaatt caacctggag caaccagacc cagaacatcc agtcctggag caaccactcc 660 tggaacactc agacctggtg cacccaatcc tggaacaatc aggcctggaa cagtcccttc 720 tataactgtg gagaggaatc tctgcagtcc tgcatgcagt tccagccaaa ttctcctgcc 780 agtgacttgg aggctgcctt ggaagctgct ggggaaggcc ttaatgtaat acagcagacc 840 actaggtatt ttagtactcc acaaaccatg gatttattcc taaactactc catgaacatg 900 caacctgaag acgtgtga 918 <210> 2 <211> 305 <212> PRT <213> Human <400> 2 Met Ser Val Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe Glu Ala 1 5 10 15 Ser Asp Cys Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly Pro Glu 20 25 30 Glu Asn Tyr Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro His Thr 35 40 45 Glu Thr Val Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile Gln Asp 50 55 60 Ser Pro Asp Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr Ser Ala 65 70 75 80 Glu Asn Ser Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys Lys Gln 85 90 95 Lys Thr Arg Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu Asn Asp 100 105 110 Arg Phe Gln Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln Glu Leu 115 120 125 Ser Asn Ile Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp Phe Gln 130 135 140 Asn Gln Arg Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp Pro Lys 145 150 155 160 Asn Ser Asn Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr Pro Ser 165 170 175 Leu Tyr Ser Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr Gly Asn 180 185 190 Leu Pro Met Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp Ser Asn 195 200 205 Gln Thr Gln Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn Thr Gln 210 215 220 Thr Trp Cys Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser Pro Phe 225 230 235 240 Tyr Asn Cys Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe Gln Pro 245 250 255 Asn Ser Pro Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala Gly Glu 260 265 270 Gly Leu Asn Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr Pro Gln 275 280 285 Thr Met Asp Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro Glu Asp 290 295 300 Val 305 <210> 3 <211> 1083 <212> DNA <213> Human <400> 3 atggcgggac acctggcttc ggatttcgcc ttctcgcccc ctccaggtgg tggaggtgat 60 gggccagggg ggccggagcc gggctgggtt gatcctcgga cctggctaag cttccaaggc 120 cctcctggag ggccaggaat cgggccgggg gttgggccag gctctgaggt gtgggggatt 180 cccccatgcc ccccgccgta tgagttctgt ggggggatgg cgtactgtgg gccccaggtt 240 ggagtggggc tagtgcccca aggcggcttg gagacctctc agcctgaggg cgaagcagga 300 gtcggggtgg agagcaactc cgatggggcc tccccggagc cctgcaccgt cacccctggt 360 gccgtgaagc tggagaagga gaagctggag caaaacccgg aggagtccca ggacatcaaa 420 gctctgcaga aagaactcga gcaatttgcc aagctcctga agcagaagag gatcaccctg 480 ggatatacac aggccgatgt ggggctcacc ctgggggttc tatttgggaa ggtattcagc 540 caaacgacca tctgccgctt tgaggctctg cagcttagct tcaagaacat gtgtaagctg 600 cggcccttgc tgcagaagtg ggtggaggaa gctgacaaca atgaaaatct tcaggagata 660 tgcaaagcag aaaccctcgt gcaggcccga aagagaaagc gaaccagtat cgagaaccga 720 gtgagaggca acctggagaa tttgttcctg cagtgcccga aacccacact gcagcagatc 780 agccacatcg cccagcagct tgggctcgag aaggatgtgg tccgagtgtg gttctgtaac 840 cggcgccaga agggcaagcg atcaagcagc gactatgcac aacgagagga ttttgaggct 900 gctgggtctc ctttctcagg gggaccagtg tcctttcctc tggccccagg gccccatttt 960 ggtaccccag gctatgggag ccctcacttc actgcactgt actcctcggt ccctttccct 1020 gagggggaag cctttccccc tgtctccgtc accactctgg gctctcccat gcattcaaac 1080 tga 1083 <210> 4 <211> 360 <212> PRT <213> Human <400> 4 Met Ala Gly His Leu Ala Ser Asp Phe Ala Phe Ser Pro Pro Pro Gly 1 5 10 15 Gly Gly Gly Asp Gly Pro Gly Gly Pro Glu Pro Gly Trp Val Asp Pro 20 25 30 Arg Thr Trp Leu Ser Phe Gln Gly Pro Pro Gly Gly Pro Gly Ile Gly 35 40 45 Pro Gly Val Gly Pro Gly Ser Glu Val Trp Gly Ile Pro Pro Cys Pro 50 55 60 Pro Pro Tyr Glu Phe Cys Gly Gly Met Ala Tyr Cys Gly Pro Gln Val 65 70 75 80 Gly Val Gly Leu Val Pro Gln Gly Gly Leu Glu Thr Ser Gln Pro Glu 85 90 95 Gly Glu Ala Gly Val Gly Val Glu Ser Asn Ser Asp Gly Ala Ser Pro 100 105 110 Glu Pro Cys Thr Val Thr Pro Gly Ala Val Lys Leu Glu Lys Glu Lys 115 120 125 Leu Glu Gln Asn Pro Glu Glu Ser Gln Asp Ile Lys Ala Leu Gln Lys 130 135 140 Glu Leu Glu Gln Phe Ala Lys Leu Leu Lys Gln Lys Arg Ile Thr Leu 145 150 155 160 Gly Tyr Thr Gln Ala Asp Val Gly Leu Thr Leu Gly Val Leu Phe Gly 165 170 175 Lys Val Phe Ser Gln Thr Thr Ile Cys Arg Phe Glu Ala Leu Gln Leu 180 185 190 Ser Phe Lys Asn Met Cys Lys Leu Arg Pro Leu Leu Gln Lys Trp Val 195 200 205 Glu Glu Ala Asp Asn Asn Glu Asn Leu Gln Glu Ile Cys Lys Ala Glu 210 215 220 Thr Leu Val Gln Ala Arg Lys Arg Lys Arg Thr Ser Ile Glu Asn Arg 225 230 235 240 Val Arg Gly Asn Leu Glu Asn Leu Phe Leu Gln Cys Pro Lys Pro Thr 245 250 255 Leu Gln Gln Ile Ser His Ile Ala Gln Gln Leu Gly Leu Glu Lys Asp 260 265 270 Val Val Arg Val Trp Phe Cys Asn Arg Arg Gln Lys Gly Lys Arg Ser 275 280 285 Ser Ser Asp Tyr Ala Gln Arg Glu Asp Phe Glu Ala Ala Gly Ser Pro 290 295 300 Phe Ser Gly Gly Pro Val Ser Phe Pro Leu Ala Pro Gly Pro His Phe 305 310 315 320 Gly Thr Pro Gly Tyr Gly Ser Pro His Phe Thr Ala Leu Tyr Ser Ser 325 330 335 Val Pro Phe Pro Glu Gly Glu Ala Phe Pro Pro Val Ser Val Thr Thr 340 345 350 Leu Gly Ser Pro Met His Ser Asn 355 360 <210> 5 <211> 954 <212> DNA <213> Human <400> 5 atgtacaaca tgatggagac ggagctgaag ccgccgggcc cgcagcaaac ttcggggggc 60 ggcggcggca actccaccgc ggcggcggcc ggcggcaacc agaaaaacag cccggaccgc 120 gtcaagcggc ccatgaatgc cttcatggtg tggtcccgcg ggcagcggcg caagatggcc 180 caggagaacc ccaagatgca caactcggag atcagcaagc gcctgggcgc cgagtggaaa 240 cttttgtcgg agacggagaa gcggccgttc atcgacgagg ctaagcggct gcgagcgctg 300 cacatgaagg agcacccgga ttataaatac cggccccggc ggaaaaccaa gacgctcatg 360 aagaaggata agtacacgct gcccggcggg ctgctggccc ccggcggcaa tagcatggcg 420 agcggggtcg gggtgggcgc cggcctgggc gcgggcgtga accagcgcat ggacagttac 480 gcgcacatga acggctggag caacggcagc tacagcatga tgcaggacca gctgggctac 540 ccgcagcacc cgggcctcaa tgcgcacggc gcagcgcaga tgcagcccat gcaccgctac 600 gacgtgagcg ccctgcagta caactccatg accagctcgc agacctacat gaacggctcg 660 cccacctaca gcatgtccta ctcgcagcag ggcacccctg gcatggctct tggctccatg 720 ggttcggtgg tcaagtccga ggccagctcc agcccccctg tggttacctc ttcctcccac 780 tccagggcgc cctgccaggc cggggacctc cgggacatga tcagcatgta tctccccggc 840 gccgaggtgc cggaacccgc cgcccccagc agacttcaca tgtcccagca ctaccagagc 900 ggcccggtgc ccggcacggc cattaacggc acactgcccc tctcacacat gtga 954 <210> 6 <211> 317 <212> PRT <213> Human <400> 6 Met Tyr Asn Met Met Glu Thr Glu Leu Lys Pro Pro Gly Pro Gln Gln 1 5 10 15 Thr Ser Gly Gly Gly Gly Gly Asn Ser Thr Ala Ala Ala Ala Gly Gly 20 25 30 Asn Gln Lys Asn Ser Pro Asp Arg Val Lys Arg Pro Met Asn Ala Phe 35 40 45 Met Val Trp Ser Arg Gly Gln Arg Arg Lys Met Ala Gln Glu Asn Pro 50 55 60 Lys Met His Asn Ser Glu Ile Ser Lys Arg Leu Gly Ala Glu Trp Lys 65 70 75 80 Leu Leu Ser Glu Thr Glu Lys Arg Pro Phe Ile Asp Glu Ala Lys Arg 85 90 95 Leu Arg Ala Leu His Met Lys Glu His Pro Asp Tyr Lys Tyr Arg Pro 100 105 110 Arg Arg Lys Thr Lys Thr Leu Met Lys Lys Asp Lys Tyr Thr Leu Pro 115 120 125 Gly Gly Leu Leu Ala Pro Gly Gly Asn Ser Met Ala Ser Gly Val Gly 130 135 140 Val Gly Ala Gly Leu Gly Ala Gly Val Asn Gln Arg Met Asp Ser Tyr 145 150 155 160 Ala His Met Asn Gly Trp Ser Asn Gly Ser Tyr Ser Met Met Gln Asp 165 170 175 Gln Leu Gly Tyr Pro Gln His Pro Gly Leu Asn Ala His Gly Ala Ala 180 185 190 Gln Met Gln Pro Met His Arg Tyr Asp Val Ser Ala Leu Gln Tyr Asn 195 200 205 Ser Met Thr Ser Ser Gln Thr Tyr Met Asn Gly Ser Pro Thr Tyr Ser 210 215 220 Met Ser Tyr Ser Gln Gln Gly Thr Pro Gly Met Ala Leu Gly Ser Met 225 230 235 240 Gly Ser Val Val Lys Ser Glu Ala Ser Ser Ser Pro Pro Val Val Thr 245 250 255 Ser Ser Ser His Ser Arg Ala Pro Cys Gln Ala Gly Asp Leu Arg Asp 260 265 270 Met Ile Ser Met Tyr Leu Pro Gly Ala Glu Val Pro Glu Pro Ala Ala 275 280 285 Pro Ser Arg Leu His Met Ser Gln His Tyr Gln Ser Gly Pro Val Pro 290 295 300 Gly Thr Ala Ile Asn Gly Thr Leu Pro Leu Ser His Met 305 310 315 <210> 7 <211> 1413 <212> DNA <213> Human <400> 7 atggctgtca gcgacgcgct gctcccatct ttctccacgt tcgcgtctgg cccggcggga 60 agggagaaga cactgcgtca agcaggtgcc ccgaataacc gctggcggga ggagctctcc 120 cacatgaagc gacttccccc agtgcttccc ggccgcccct atgacctggc ggcggcgacc 180 gtggccacag acctggagag cggcggagcc ggtgcggctt gcggcggtag caacctggcg 240 cccctacctc ggagagagac cgaggagttc aacgatctcc tggacctgga ctttattctc 300 tccaattcgc tgacccatcc tccggagtca gtggccgcca ccgtgtcctc gtcagcgtca 360 gcctcctctt cgtcgtcgcc gtcgagcagc ggccctgcca gcgcgccctc cacctgcagc 420 ttcacctatc cgatccgggc cgggaacgac ccgggcgtgg cgccgggcgg cacgggcgga 480 ggcctcctct atggcaggga gtccgctccc cctccgacgg ctcccttcaa cctggcggac 540 atcaacgacg tgagcccctc gggcggcttc gtggccgagc tcctgcggcc agaattggac 600 ccggtgtaca ttccgccgca gcagccgcag ccgccaggtg gcgggctgat gggcaagttc 660 gtgctgaagg cgtcgctgag cgcccctggc agcgagtacg gcagcccgtc ggtcatcagc 720 gtcagcaaag gcagccctga cggcagccac ccggtggtgg tggcgcccta caacggcggg 780 ccgccgcgca cgtgccccaa gatcaagcag gaggcggtct cttcgtgcac ccacttgggc 840 gctggacccc ctctcagcaa tggccaccgg ccggctgcac acgacttccc cctggggcgg 900 cagctcccca gcaggactac cccgaccctg ggtcttgagg aagtgctgag cagcagggac 960 tgtcaccctg ccctgccgct tcctcccggc ttccatcccc acccggggcc caattaccca 1020 tccttcctgc ccgatcagat gcagccgcaa gtcccgccgc tccattacca agagctcatg 1080 ccacccggtt cctgcatgcc agaggagccc aagccaaaga ggggaagacg atcgtggccc 1140 cggaaaagga ccgccaccca cacttgtgat tacgcgggct gcggcaaaac ctacacaaag 1200 agttcccatc tcaaggcaca cctgcgaacc cacacaggtg agaaacctta ccactgtgac 1260 tgggacggct gtggatggaa attcgcccgc tcagatgaac tgaccaggca ctaccgtaaa 1320 cacacggggc accgcccgtt ccagtgccaa aaatgcgacc gagcattttc caggtcggac 1380 cacctcgcct tacacatgaa gaggcatttt taa 1413 <210> 8 <211> 470 <212> PRT <213> Human <400> 8 Met Ala Val Ser Asp Ala Leu Leu Pro Ser Phe Ser Thr Phe Ala Ser 1 5 10 15 Gly Pro Ala Gly Arg Glu Lys Thr Leu Arg Gln Ala Gly Ala Pro Asn 20 25 30 Asn Arg Trp Arg Glu Glu Leu Ser His Met Lys Arg Leu Pro Pro Val 35 40 45 Leu Pro Gly Arg Pro Tyr Asp Leu Ala Ala Ala Thr Val Ala Thr Asp 50 55 60 Leu Glu Ser Gly Gly Ala Gly Ala Ala Cys Gly Gly Ser Asn Leu Ala 65 70 75 80 Pro Leu Pro Arg Arg Glu Thr Glu Glu Phe Asn Asp Leu Leu Asp Leu 85 90 95 Asp Phe Ile Leu Ser Asn Ser Leu Thr His Pro Pro Glu Ser Val Ala 100 105 110 Ala Thr Val Ser Ser Ser Ala Ser Ala Ser Ser Ser Ser Ser Pro Ser 115 120 125 Ser Ser Gly Pro Ala Ser Ala Pro Ser Thr Cys Ser Phe Thr Tyr Pro 130 135 140 Ile Arg Ala Gly Asn Asp Pro Gly Val Ala Pro Gly Gly Thr Gly Gly 145 150 155 160 Gly Leu Leu Tyr Gly Arg Glu Ser Ala Pro Pro Pro Thr Ala Pro Phe 165 170 175 Asn Leu Ala Asp Ile Asn Asp Val Ser Pro Ser Gly Gly Phe Val Ala 180 185 190 Glu Leu Leu Arg Pro Glu Leu Asp Pro Val Tyr Ile Pro Pro Gln Gln 195 200 205 Pro Gln Pro Pro Gly Gly Gly Leu Met Gly Lys Phe Val Leu Lys Ala 210 215 220 Ser Leu Ser Ala Pro Gly Ser Glu Tyr Gly Ser Pro Ser Val Ile Ser 225 230 235 240 Val Ser Lys Gly Ser Pro Asp Gly Ser His Pro Val Val Val Ala Pro 245 250 255 Tyr Asn Gly Gly Pro Pro Arg Thr Cys Pro Lys Ile Lys Gln Glu Ala 260 265 270 Val Ser Ser Cys Thr His Leu Gly Ala Gly Pro Pro Leu Ser Asn Gly 275 280 285 His Arg Pro Ala Ala His Asp Phe Pro Leu Gly Arg Gln Leu Pro Ser 290 295 300 Arg Thr Thr Pro Thr Leu Gly Leu Glu Glu Val Leu Ser Ser Arg Asp 305 310 315 320 Cys His Pro Ala Leu Pro Leu Pro Pro Gly Phe His Pro His Pro Gly 325 330 335 Pro Asn Tyr Pro Ser Phe Leu Pro Asp Gln Met Gln Pro Gln Val Pro 340 345 350 Pro Leu His Tyr Gln Glu Leu Met Pro Pro Gly Ser Cys Met Pro Glu 355 360 365 Glu Pro Lys Pro Lys Arg Gly Arg Arg Ser Trp Pro Arg Lys Arg Thr 370 375 380 Ala Thr His Thr Cys Asp Tyr Ala Gly Cys Gly Lys Thr Tyr Thr Lys 385 390 395 400 Ser Ser His Leu Lys Ala His Leu Arg Thr His Thr Gly Glu Lys Pro 405 410 415 Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp 420 425 430 Glu Leu Thr Arg His Tyr Arg Lys His Thr Gly His Arg Pro Phe Gln 435 440 445 Cys Gln Lys Cys Asp Arg Ala Phe Ser Arg Ser Asp His Leu Ala Leu 450 455 460 His Met Lys Arg His Phe 465 470 <210> 9 <211> 1368 <212> DNA <213> Human <400> 9 atgctggatt tttttcgggt agtggaaaac cagcagcctc ccgcgacgat gcccctcaac 60 gttagcttca ccaacaggaa ctatgacctc gactacgact cggtgcagcc gtatttctac 120 tgcgacgagg aggagaactt ctaccagcag cagcagcaga gcgagctgca gcccccggcg 180 cccagcgagg atatctggaa gaaattcgag ctgctgccca ccccgcccct gtcccctagc 240 cgccgctccg ggctctgctc gccctcctac gttgcggtca cacccttctc ccttcgggga 300 gacaacgacg gcggtggcgg gagcttctcc acggccgacc agctggagat ggtgaccgag 360 ctgctgggag gagacatggt gaaccagagt ttcatctgcg acccggacga cgagaccttc 420 atcaaaaaca tcatcatcca ggactgtatg tggagcggct tctcggccgc cgccaagctc 480 gtctcagaga agctggcctc ctaccaggct gcgcgcaaag acagcggcag cccgaacccc 540 gcccgcggcc acagcgtctg ctccacctcc agcttgtacc tgcaggatct gagcgccgcc 600 gcctcagagt gcatcgaccc ctcggtggtc ttcccctacc ctctcaacga cagcagctcg 660 cccaagtcct gcgcctcgca agactccagc gccttctctc cgtcctcgga ttctctgctc 720 tcctcgacgg agtcctcccc gcagggcagc cccgagcccc tggtgctcca tgaggagaca 780 ccgcccacca ccagcagcga ctctgaggag gaacaagaag atgaggaaga aatcgatgtt 840 gtttctgtgg aaaagaggca ggctcctggc aaaaggtcag agtctggatc accttctgct 900 ggaggccaca gcaaacctcc tcacagccca ctggtcctca agaggtgcca cgtctccaca 960 catcagcaca actacgcagc gcctccctcc actcggaagg actatcctgc tgccaagagg 1020 gtcaagttgg acagtgtcag agtcctgaga cagatcagca acaaccgaaa atgcaccagc 1080 cccaggtcct cggacaccga ggagaatgtc aagaggcgaa cacacaacgt cttggagcgc 1140 cagaggagga acgagctaaa acggagcttt tttgccctgc gtgaccagat cccggagttg 1200 gaaaacaatg aaaaggcccc caaggtagtt atccttaaaa aagccacagc atacatcctg 1260 tccgtccaag cagaggagca aaagctcatt tctgaagagg acttgttgcg gaaacgacga 1320 gaacagttga aacacaaact tgaacagcta cggaactctt gtgcgtaa 1368 <210> 10 <211> 455 <212> PRT <213> Human <400> 10 Met Leu Asp Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro Ala Thr 1 5 10 15 Met Pro Leu Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu Asp Tyr 20 25 30 Asp Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn Phe Tyr 35 40 45 Gln Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser Glu Asp 50 55 60 Ile Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser Pro Ser 65 70 75 80 Arg Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr Pro Phe 85 90 95 Ser Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser Thr Ala 100 105 110 Asp Gln Leu Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met Val Asn 115 120 125 Gln Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys Asn Ile 130 135 140 Ile Ile Gln Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala Lys Leu 145 150 155 160 Val Ser Glu Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp Ser Gly 165 170 175 Ser Pro Asn Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser Ser Leu 180 185 190 Tyr Leu Gln Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp Pro Ser 195 200 205 Val Val Phe Pro Tyr Pro Leu Asn Asp Ser Ser Ser Pro Lys Ser Cys 210 215 220 Ala Ser Gln Asp Ser Ser Ala Phe Ser Pro Ser Ser Asp Ser Leu Leu 225 230 235 240 Ser Ser Thr Glu Ser Ser Pro Gln Gly Ser Pro Glu Pro Leu Val Leu 245 250 255 His Glu Glu Thr Pro Pro Thr Thr Ser Ser Asp Ser Glu Glu Glu Gln 260 265 270 Glu Asp Glu Glu Glu Ile Asp Val Val Ser Val Glu Lys Arg Gln Ala 275 280 285 Pro Gly Lys Arg Ser Glu Ser Gly Ser Pro Ser Ala Gly Gly His Ser 290 295 300 Lys Pro Pro His Ser Pro Leu Val Leu Lys Arg Cys His Val Ser Thr 305 310 315 320 His Gln His Asn Tyr Ala Ala Pro Pro Ser Thr Arg Lys Asp Tyr Pro 325 330 335 Ala Ala Lys Arg Val Lys Leu Asp Ser Val Arg Val Leu Arg Gln Ile 340 345 350 Ser Asn Asn Arg Lys Cys Thr Ser Pro Arg Ser Ser Asp Thr Glu Glu 355 360 365 Asn Val Lys Arg Arg Thr His Asn Val Leu Glu Arg Gln Arg Arg Asn 370 375 380 Glu Leu Lys Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro Glu Leu 385 390 395 400 Glu Asn Asn Glu Lys Ala Pro Lys Val Val Ile Leu Lys Lys Ala Thr 405 410 415 Ala Tyr Ile Leu Ser Val Gln Ala Glu Glu Gln Lys Leu Ile Ser Glu 420 425 430 Glu Asp Leu Leu Arg Lys Arg Arg Glu Gln Leu Lys His Lys Leu Glu 435 440 445 Gln Leu Arg Asn Ser Cys Ala 450 455 <210> 11 <211> 630 <212> DNA <213> Human <400> 11 atgggctccg tgtccaacca gcagtttgca ggtggctgcg ccaaggcggc agaagaggcg 60 cccgaggagg cgccggagga cgcggcccgg gcggcggacg agcctcagct gctgcacggt 120 gcgggcatct gtaagtggtt caacgtgcgc atggggttcg gcttcctgtc catgaccgcc 180 cgcgccgggg tcgcgctcga ccccccagtg gatgtctttg tgcaccagag taagctgcac 240 atggaagggt tccggagctt gaaggagggt gaggcagtgg agttcacctt taagaagtca 300 gccaagggtc tggaatccat ccgtgtcacc ggacctggtg gagtattctg tattgggagt 360 gagaggcggc caaaaggaaa gagcatgcag aagcgcagat caaaaggaga caggtgctac 420 aactgtggag gtctagatca tcatgccaag gaatgcaagc tgccacccca gcccaagaag 480 tgccacttct gccagagcat cagccatatg gtagcctcat gtccgctgaa ggcccagcag 540 ggccctagtg cacagggaaa gccaacctac tttcgagagg aagaagaaga aatccacagc 600 cctaccctgc tcccggaggc acagaattga 630 <210> 12 <211> 209 <212> PRT <213> Human <400> 12 Met Gly Ser Val Ser Asn Gln Gln Phe Ala Gly Gly Cys Ala Lys Ala 1 5 10 15 Ala Glu Glu Ala Pro Glu Glu Ala Pro Glu Asp Ala Ala Arg Ala Ala 20 25 30 Asp Glu Pro Gln Leu Leu His Gly Ala Gly Ile Cys Lys Trp Phe Asn 35 40 45 Val Arg Met Gly Phe Gly Phe Leu Ser Met Thr Ala Arg Ala Gly Val 50 55 60 Ala Leu Asp Pro Pro Val Asp Val Phe Val His Gln Ser Lys Leu His 65 70 75 80 Met Glu Gly Phe Arg Ser Leu Lys Glu Gly Glu Ala Val Glu Phe Thr 85 90 95 Phe Lys Lys Ser Ala Lys Gly Leu Glu Ser Ile Arg Val Thr Gly Pro 100 105 110 Gly Gly Val Phe Cys Ile Gly Ser Glu Arg Arg Pro Lys Gly Lys Ser 115 120 125 Met Gln Lys Arg Arg Ser Lys Gly Asp Arg Cys Tyr Asn Cys Gly Gly 130 135 140 Leu Asp His His Ala Lys Glu Cys Lys Leu Pro Pro Gln Pro Lys Lys 145 150 155 160 Cys His Phe Cys Gln Ser Ile Ser His Met Val Ala Ser Cys Pro Leu 165 170 175 Lys Ala Gln Gln Gly Pro Ser Ala Gln Gly Lys Pro Thr Tyr Phe Arg 180 185 190 Glu Glu Glu Glu Glu Ile His Ser Pro Thr Leu Leu Pro Glu Ala Gln 195 200 205 Asn <210> 13 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> SV40 large T antigen-derived NLS cDNA Sequence <400> 13 aagaagaaga ggaag 15 <210> 14 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> SV40 large T antigen-derived NLS Amino Acid Sequence <400> 14 Lys Lys Lys Arg Lys 1 5 <210> 15 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> JO-84 MTD cDNA Sequence <400> 15 ctggtggcgg cgctgctggc ggtgctg 27 <210> 16 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> JO-84 MTD Amino Acid Sequence <400> 16 Leu Val Ala Ala Leu Leu Ala Val Leu 1 5 <210> 17 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> JO-86 MTD cDNA Sequence <400> 17 ctggcggtgc tggcggcggc gccg 24 <210> 18 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> JO-86 MTD Amino Acid Sequence <400> 18 Leu Ala Val Leu Ala Ala Ala Pro 1 5 <210> 19 <211> 933 <212> DNA <213> Artificial Sequence <220> <223> NLS-Nanog cDNA Sequence <400> 19 atgaagaaga agaggaagag tgtggatcca gcttgtcccc aaagcttgcc ttgctttgaa 60 gcatccgact gtaaagaatc ttcacctatg cctgtgattt gtgggcctga agaaaactat 120 ccatccttgc aaatgtcttc tgctgagatg cctcacacgg agactgtctc tcctcttccc 180 tcctccatgg atctgcttat tcaggacagc cctgattctt ccaccagtcc caaaggcaaa 240 caacccactt ctgcagagaa tagtgtcgca aaaaaggaag acaaggtccc agtcaagaaa 300 cagaagacca gaactgtgtt ctcttccacc cagctgtgtg tactcaatga tagatttcag 360 agacagaaat acctcagcct ccagcagatg caagaactct ccaacatcct gaacctcagc 420 tacaaacagg tgaagacctg gttccagaac cagagaatga aatctaagag gtggcagaaa 480 aacaactggc cgaagaatag caatggtgtg acgcagaagg cctcagcacc tacctacccc 540 agcctctact cttcctacca ccagggatgc ctggtgaacc cgactgggaa ccttccaatg 600 tggagcaacc agacctggaa caattcaacc tggagcaacc agacccagaa catccagtcc 660 tggagcaacc actcctggaa cactcagacc tggtgcaccc aatcctggaa caatcaggcc 720 tggaacagtc ccttctataa ctgtggagag gaatctctgc agtcctgcat gcagttccag 780 ccaaattctc ctgccagtga cttggaggct gctttggaag ctgctgggga aggccttaat 840 gtaatacagc agaccactag gtattttagt actccacaaa ccatggattt attcctaaac 900 tactccatga acatgcaacc tgaagacgtg tga 933 <210> 20 <211> 310 <212> PRT <213> Artificial Sequence <220> <223> NLS-Nanog Amino Acid Sequence <400> 20 Met Lys Lys Lys Arg Lys Ser Val Asp Pro Ala Cys Pro Gln Ser Leu 1 5 10 15 Pro Cys Phe Glu Ala Ser Asp Cys Lys Glu Ser Ser Pro Met Pro Val 20 25 30 Ile Cys Gly Pro Glu Glu Asn Tyr Pro Ser Leu Gln Met Ser Ser Ala 35 40 45 Glu Met Pro His Thr Glu Thr Val Ser Pro Leu Pro Ser Ser Met Asp 50 55 60 Leu Leu Ile Gln Asp Ser Pro Asp Ser Ser Thr Ser Pro Lys Gly Lys 65 70 75 80 Gln Pro Thr Ser Ala Glu Asn Ser Val Ala Lys Lys Glu Asp Lys Val 85 90 95 Pro Val Lys Lys Gln Lys Thr Arg Thr Val Phe Ser Ser Thr Gln Leu 100 105 110 Cys Val Leu Asn Asp Arg Phe Gln Arg Gln Lys Tyr Leu Ser Leu Gln 115 120 125 Gln Met Gln Glu Leu Ser Asn Ile Leu Asn Leu Ser Tyr Lys Gln Val 130 135 140 Lys Thr Trp Phe Gln Asn Gln Arg Met Lys Ser Lys Arg Trp Gln Lys 145 150 155 160 Asn Asn Trp Pro Lys Asn Ser Asn Gly Val Thr Gln Lys Ala Ser Ala 165 170 175 Pro Thr Tyr Pro Ser Leu Tyr Ser Ser Tyr His Gln Gly Cys Leu Val 180 185 190 Asn Pro Thr Gly Asn Leu Pro Met Trp Ser Asn Gln Thr Trp Asn Asn 195 200 205 Ser Thr Trp Ser Asn Gln Thr Gln Asn Ile Gln Ser Trp Ser Asn His 210 215 220 Ser Trp Asn Thr Gln Thr Trp Cys Thr Gln Ser Trp Asn Asn Gln Ala 225 230 235 240 Trp Asn Ser Pro Phe Tyr Asn Cys Gly Glu Glu Ser Leu Gln Ser Cys 245 250 255 Met Gln Phe Gln Pro Asn Ser Pro Ala Ser Asp Leu Glu Ala Ala Leu 260 265 270 Glu Ala Ala Gly Glu Gly Leu Asn Val Ile Gln Gln Thr Thr Arg Tyr 275 280 285 Phe Ser Thr Pro Gln Thr Met Asp Leu Phe Leu Asn Tyr Ser Met Asn 290 295 300 Met Gln Pro Glu Asp Val 305 310 <210> 21 <211> 960 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Nanog cDNA Sequence <400> 21 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgagtgt ggatccagct 60 tgtccccaaa gcttgccttg ctttgaagca tccgactgta aagaatcttc acctatgcct 120 gtgatttgtg ggcctgaaga aaactatcca tccttgcaaa tgtcttctgc tgagatgcct 180 cacacggaga ctgtctctcc tcttccctcc tccatggatc tgcttattca ggacagccct 240 gattcttcca ccagtcccaa aggcaaacaa cccacttctg cagagaatag tgtcgcaaaa 300 aaggaagaca aggtcccagt caagaaacag aagaccagaa ctgtgttctc ttccacccag 360 ctgtgtgtac tcaatgatag atttcagaga cagaaatacc tcagcctcca gcagatgcaa 420 gaactctcca acatcctgaa cctcagctac aaacaggtga agacctggtt ccagaaccag 480 agaatgaaat ctaagaggtg gcagaaaaac aactggccga agaatagcaa tggtgtgacg 540 cagaaggcct cagcacctac ctaccccagc ctctactctt cctaccacca gggatgcctg 600 gtgaacccga ctgggaacct tccaatgtgg agcaaccaga cctggaacaa ttcaacctgg 660 agcaaccaga cccagaacat ccagtcctgg agcaaccact cctggaacac tcagacctgg 720 tgcacccaat cctggaacaa tcaggcctgg aacagtccct tctataactg tggagaggaa 780 tctctgcagt cctgcatgca gttccagcca aattctcctg ccagtgactt ggaggctgct 840 ttggaagctg ctggggaagg ccttaatgta atacagcaga ccactaggta ttttagtact 900 ccacaaacca tggatttatt cctaaactac tccatgaaca tgcaacctga agacgtgtga 960 960 <210> 22 <211> 319 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Nanog Amino Acid Sequence <400> 22 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Ser 1 5 10 15 Val Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe Glu Ala Ser Asp 20 25 30 Cys Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly Pro Glu Glu Asn 35 40 45 Tyr Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro His Thr Glu Thr 50 55 60 Val Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile Gln Asp Ser Pro 65 70 75 80 Asp Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr Ser Ala Glu Asn 85 90 95 Ser Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys Lys Gln Lys Thr 100 105 110 Arg Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu Asn Asp Arg Phe 115 120 125 Gln Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln Glu Leu Ser Asn 130 135 140 Ile Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp Phe Gln Asn Gln 145 150 155 160 Arg Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp Pro Lys Asn Ser 165 170 175 Asn Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr Pro Ser Leu Tyr 180 185 190 Ser Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr Gly Asn Leu Pro 195 200 205 Met Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp Ser Asn Gln Thr 210 215 220 Gln Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn Thr Gln Thr Trp 225 230 235 240 Cys Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser Pro Phe Tyr Asn 245 250 255 Cys Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe Gln Pro Asn Ser 260 265 270 Pro Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala Gly Glu Gly Leu 275 280 285 Asn Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr Pro Gln Thr Met 290 295 300 Asp Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro Glu Asp Val 305 310 315 <210> 23 <211> 960 <212> DNA <213> Artificial Sequence <220> <223> NLS-Nanog-JO-84 MTD cDNA Sequence <400> 23 atgaagaaga agaggaagag tgtggatcca gcttgtcccc aaagcttgcc ttgctttgaa 60 gcatccgact gtaaagaatc ttcacctatg cctgtgattt gtgggcctga agaaaactat 120 ccatccttgc aaatgtcttc tgctgagatg cctcacacgg agactgtctc tcctcttccc 180 tcctccatgg atctgcttat tcaggacagc cctgattctt ccaccagtcc caaaggcaaa 240 caacccactt ctgcagagaa tagtgtcgca aaaaaggaag acaaggtccc agtcaagaaa 300 cagaagacca gaactgtgtt ctcttccacc cagctgtgtg tactcaatga tagatttcag 360 agacagaaat acctcagcct ccagcagatg caagaactct ccaacatcct gaacctcagc 420 tacaaacagg tgaagacctg gttccagaac cagagaatga aatctaagag gtggcagaaa 480 aacaactggc cgaagaatag caatggtgtg acgcagaagg cctcagcacc tacctacccc 540 agcctctact cttcctacca ccagggatgc ctggtgaacc cgactgggaa ccttccaatg 600 tggagcaacc agacctggaa caattcaacc tggagcaacc agacccagaa catccagtcc 660 tggagcaacc actcctggaa cactcagacc tggtgcaccc aatcctggaa caatcaggcc 720 tggaacagtc ccttctataa ctgtggagag gaatctctgc agtcctgcat gcagttccag 780 ccaaattctc ctgccagtga cttggaggct gctttggaag ctgctgggga aggccttaat 840 gtaatacagc agaccactag gtattttagt actccacaaa ccatggattt attcctaaac 900 tactccatga acatgcaacc tgaagacgtg ctggtggcgg cgctgctggc ggtgctgtga 960 960 <210> 24 <211> 319 <212> PRT <213> Artificial Sequence <220> <223> NLS-Nanog-JO-84 MTD Amino Acid Sequence <400> 24 Met Lys Lys Lys Arg Lys Ser Val Asp Pro Ala Cys Pro Gln Ser Leu 1 5 10 15 Pro Cys Phe Glu Ala Ser Asp Cys Lys Glu Ser Ser Pro Met Pro Val 20 25 30 Ile Cys Gly Pro Glu Glu Asn Tyr Pro Ser Leu Gln Met Ser Ser Ala 35 40 45 Glu Met Pro His Thr Glu Thr Val Ser Pro Leu Pro Ser Ser Met Asp 50 55 60 Leu Leu Ile Gln Asp Ser Pro Asp Ser Ser Thr Ser Pro Lys Gly Lys 65 70 75 80 Gln Pro Thr Ser Ala Glu Asn Ser Val Ala Lys Lys Glu Asp Lys Val 85 90 95 Pro Val Lys Lys Gln Lys Thr Arg Thr Val Phe Ser Ser Thr Gln Leu 100 105 110 Cys Val Leu Asn Asp Arg Phe Gln Arg Gln Lys Tyr Leu Ser Leu Gln 115 120 125 Gln Met Gln Glu Leu Ser Asn Ile Leu Asn Leu Ser Tyr Lys Gln Val 130 135 140 Lys Thr Trp Phe Gln Asn Gln Arg Met Lys Ser Lys Arg Trp Gln Lys 145 150 155 160 Asn Asn Trp Pro Lys Asn Ser Asn Gly Val Thr Gln Lys Ala Ser Ala 165 170 175 Pro Thr Tyr Pro Ser Leu Tyr Ser Ser Tyr His Gln Gly Cys Leu Val 180 185 190 Asn Pro Thr Gly Asn Leu Pro Met Trp Ser Asn Gln Thr Trp Asn Asn 195 200 205 Ser Thr Trp Ser Asn Gln Thr Gln Asn Ile Gln Ser Trp Ser Asn His 210 215 220 Ser Trp Asn Thr Gln Thr Trp Cys Thr Gln Ser Trp Asn Asn Gln Ala 225 230 235 240 Trp Asn Ser Pro Phe Tyr Asn Cys Gly Glu Glu Ser Leu Gln Ser Cys 245 250 255 Met Gln Phe Gln Pro Asn Ser Pro Ala Ser Asp Leu Glu Ala Ala Leu 260 265 270 Glu Ala Ala Gly Glu Gly Leu Asn Val Ile Gln Gln Thr Thr Arg Tyr 275 280 285 Phe Ser Thr Pro Gln Thr Met Asp Leu Phe Leu Asn Tyr Ser Met Asn 290 295 300 Met Gln Pro Glu Asp Val Leu Val Ala Ala Leu Leu Ala Val Leu 305 310 315 <210> 25 <211> 987 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Nanog-JO-84 MTD cDNA Sequence <400> 25 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgagtgt ggatccagct 60 tgtccccaaa gcttgccttg ctttgaagca tccgactgta aagaatcttc acctatgcct 120 gtgatttgtg ggcctgaaga aaactatcca tccttgcaaa tgtcttctgc tgagatgcct 180 cacacggaga ctgtctctcc tcttccctcc tccatggatc tgcttattca ggacagccct 240 gattcttcca ccagtcccaa aggcaaacaa cccacttctg cagagaatag tgtcgcaaaa 300 aaggaagaca aggtcccagt caagaaacag aagaccagaa ctgtgttctc ttccacccag 360 ctgtgtgtac tcaatgatag atttcagaga cagaaatacc tcagcctcca gcagatgcaa 420 gaactctcca acatcctgaa cctcagctac aaacaggtga agacctggtt ccagaaccag 480 agaatgaaat ctaagaggtg gcagaaaaac aactggccga agaatagcaa tggtgtgacg 540 cagaaggcct cagcacctac ctaccccagc ctctactctt cctaccacca gggatgcctg 600 gtgaacccga ctgggaacct tccaatgtgg agcaaccaga cctggaacaa ttcaacctgg 660 agcaaccaga cccagaacat ccagtcctgg agcaaccact cctggaacac tcagacctgg 720 tgcacccaat cctggaacaa tcaggcctgg aacagtccct tctataactg tggagaggaa 780 tctctgcagt cctgcatgca gttccagcca aattctcctg ccagtgactt ggaggctgct 840 ttggaagctg ctggggaagg ccttaatgta atacagcaga ccactaggta ttttagtact 900 ccacaaacca tggatttatt cctaaactac tccatgaaca tgcaacctga agacgtgctg 960 gtggcggcgc tgctggcggt gctgtga 987 <210> 26 <211> 328 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Nanog-JO-84 MTD Amino Acid Sequence <400> 26 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Ser 1 5 10 15 Val Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe Glu Ala Ser Asp 20 25 30 Cys Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly Pro Glu Glu Asn 35 40 45 Tyr Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro His Thr Glu Thr 50 55 60 Val Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile Gln Asp Ser Pro 65 70 75 80 Asp Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr Ser Ala Glu Asn 85 90 95 Ser Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys Lys Gln Lys Thr 100 105 110 Arg Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu Asn Asp Arg Phe 115 120 125 Gln Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln Glu Leu Ser Asn 130 135 140 Ile Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp Phe Gln Asn Gln 145 150 155 160 Arg Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp Pro Lys Asn Ser 165 170 175 Asn Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr Pro Ser Leu Tyr 180 185 190 Ser Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr Gly Asn Leu Pro 195 200 205 Met Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp Ser Asn Gln Thr 210 215 220 Gln Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn Thr Gln Thr Trp 225 230 235 240 Cys Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser Pro Phe Tyr Asn 245 250 255 Cys Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe Gln Pro Asn Ser 260 265 270 Pro Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala Gly Glu Gly Leu 275 280 285 Asn Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr Pro Gln Thr Met 290 295 300 Asp Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro Glu Asp Val Leu 305 310 315 320 Val Ala Ala Leu Leu Ala Val Leu 325 <210> 27 <211> 957 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Nanog cDNA Sequence <400> 27 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgagtgtgga tccagcttgt 60 ccccaaagct tgccttgctt tgaagcatcc gactgtaaag aatcttcacc tatgcctgtg 120 atttgtgggc ctgaagaaaa ctatccatcc ttgcaaatgt cttctgctga gatgcctcac 180 acggagactg tctctcctct tccctcctcc atggatctgc ttattcagga cagccctgat 240 tcttccacca gtcccaaagg caaacaaccc acttctgcag agaatagtgt cgcaaaaaag 300 gaagacaagg tcccagtcaa gaaacagaag accagaactg tgttctcttc cacccagctg 360 tgtgtactca atgatagatt tcagagacag aaatacctca gcctccagca gatgcaagaa 420 ctctccaaca tcctgaacct cagctacaaa caggtgaaga cctggttcca gaaccagaga 480 atgaaatcta agaggtggca gaaaaacaac tggccgaaga atagcaatgg tgtgacgcag 540 aaggcctcag cacctaccta ccccagcctc tactcttcct accaccaggg atgcctggtg 600 aacccgactg ggaaccttcc aatgtggagc aaccagacct ggaacaattc aacctggagc 660 aaccagaccc agaacatcca gtcctggagc aaccactcct ggaacactca gacctggtgc 720 acccaatcct ggaacaatca ggcctggaac agtcccttct ataactgtgg agaggaatct 780 ctgcagtcct gcatgcagtt ccagccaaat tctcctgcca gtgacttgga ggctgctttg 840 gaagctgctg gggaaggcct taatgtaata cagcagacca ctaggtattt tagtactcca 900 caaaccatgg atttattcct aaactactcc atgaacatgc aacctgaaga cgtgtga 957 <210> 28 <211> 318 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Nanog Amino Acid Sequence <400> 28 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Ser Val 1 5 10 15 Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe Glu Ala Ser Asp Cys 20 25 30 Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly Pro Glu Glu Asn Tyr 35 40 45 Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro His Thr Glu Thr Val 50 55 60 Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile Gln Asp Ser Pro Asp 65 70 75 80 Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr Ser Ala Glu Asn Ser 85 90 95 Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys Lys Gln Lys Thr Arg 100 105 110 Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu Asn Asp Arg Phe Gln 115 120 125 Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln Glu Leu Ser Asn Ile 130 135 140 Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp Phe Gln Asn Gln Arg 145 150 155 160 Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp Pro Lys Asn Ser Asn 165 170 175 Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr Pro Ser Leu Tyr Ser 180 185 190 Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr Gly Asn Leu Pro Met 195 200 205 Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp Ser Asn Gln Thr Gln 210 215 220 Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn Thr Gln Thr Trp Cys 225 230 235 240 Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser Pro Phe Tyr Asn Cys 245 250 255 Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe Gln Pro Asn Ser Pro 260 265 270 Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala Gly Glu Gly Leu Asn 275 280 285 Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr Pro Gln Thr Met Asp 290 295 300 Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro Glu Asp Val 305 310 315 <210> 29 <211> 957 <212> DNA <213> Artificial Sequence <220> <223> NLS-Nanog-JO-86 MTD cDNA Sequence <400> 29 atgaagaaga agaggaagag tgtggatcca gcttgtcccc aaagcttgcc ttgctttgaa 60 gcatccgact gtaaagaatc ttcacctatg cctgtgattt gtgggcctga agaaaactat 120 ccatccttgc aaatgtcttc tgctgagatg cctcacacgg agactgtctc tcctcttccc 180 tcctccatgg atctgcttat tcaggacagc cctgattctt ccaccagtcc caaaggcaaa 240 caacccactt ctgcagagaa tagtgtcgca aaaaaggaag acaaggtccc agtcaagaaa 300 cagaagacca gaactgtgtt ctcttccacc cagctgtgtg tactcaatga tagatttcag 360 agacagaaat acctcagcct ccagcagatg caagaactct ccaacatcct gaacctcagc 420 tacaaacagg tgaagacctg gttccagaac cagagaatga aatctaagag gtggcagaaa 480 aacaactggc cgaagaatag caatggtgtg acgcagaagg cctcagcacc tacctacccc 540 agcctctact cttcctacca ccagggatgc ctggtgaacc cgactgggaa ccttccaatg 600 tggagcaacc agacctggaa caattcaacc tggagcaacc agacccagaa catccagtcc 660 tggagcaacc actcctggaa cactcagacc tggtgcaccc aatcctggaa caatcaggcc 720 tggaacagtc ccttctataa ctgtggagag gaatctctgc agtcctgcat gcagttccag 780 ccaaattctc ctgccagtga cttggaggct gctttggaag ctgctgggga aggccttaat 840 gtaatacagc agaccactag gtattttagt actccacaaa ccatggattt attcctaaac 900 tactccatga acatgcaacc tgaagacgtg ctggcggtgc tggcggcggc gccgtga 957 <210> 30 <211> 318 <212> PRT <213> Artificial Sequence <220> <223> NLS-Nanog-JO-86 MTD Amino Acid Sequence <400> 30 Met Lys Lys Lys Arg Lys Ser Val Asp Pro Ala Cys Pro Gln Ser Leu 1 5 10 15 Pro Cys Phe Glu Ala Ser Asp Cys Lys Glu Ser Ser Pro Met Pro Val 20 25 30 Ile Cys Gly Pro Glu Glu Asn Tyr Pro Ser Leu Gln Met Ser Ser Ala 35 40 45 Glu Met Pro His Thr Glu Thr Val Ser Pro Leu Pro Ser Ser Met Asp 50 55 60 Leu Leu Ile Gln Asp Ser Pro Asp Ser Ser Thr Ser Pro Lys Gly Lys 65 70 75 80 Gln Pro Thr Ser Ala Glu Asn Ser Val Ala Lys Lys Glu Asp Lys Val 85 90 95 Pro Val Lys Lys Gln Lys Thr Arg Thr Val Phe Ser Ser Thr Gln Leu 100 105 110 Cys Val Leu Asn Asp Arg Phe Gln Arg Gln Lys Tyr Leu Ser Leu Gln 115 120 125 Gln Met Gln Glu Leu Ser Asn Ile Leu Asn Leu Ser Tyr Lys Gln Val 130 135 140 Lys Thr Trp Phe Gln Asn Gln Arg Met Lys Ser Lys Arg Trp Gln Lys 145 150 155 160 Asn Asn Trp Pro Lys Asn Ser Asn Gly Val Thr Gln Lys Ala Ser Ala 165 170 175 Pro Thr Tyr Pro Ser Leu Tyr Ser Ser Tyr His Gln Gly Cys Leu Val 180 185 190 Asn Pro Thr Gly Asn Leu Pro Met Trp Ser Asn Gln Thr Trp Asn Asn 195 200 205 Ser Thr Trp Ser Asn Gln Thr Gln Asn Ile Gln Ser Trp Ser Asn His 210 215 220 Ser Trp Asn Thr Gln Thr Trp Cys Thr Gln Ser Trp Asn Asn Gln Ala 225 230 235 240 Trp Asn Ser Pro Phe Tyr Asn Cys Gly Glu Glu Ser Leu Gln Ser Cys 245 250 255 Met Gln Phe Gln Pro Asn Ser Pro Ala Ser Asp Leu Glu Ala Ala Leu 260 265 270 Glu Ala Ala Gly Glu Gly Leu Asn Val Ile Gln Gln Thr Thr Arg Tyr 275 280 285 Phe Ser Thr Pro Gln Thr Met Asp Leu Phe Leu Asn Tyr Ser Met Asn 290 295 300 Met Gln Pro Glu Asp Val Leu Ala Val Leu Ala Ala Ala Pro 305 310 315 <210> 31 <211> 981 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Nanog-JO-86 MTD cDNA Sequence <400> 31 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgagtgtgga tccagcttgt 60 ccccaaagct tgccttgctt tgaagcatcc gactgtaaag aatcttcacc tatgcctgtg 120 atttgtgggc ctgaagaaaa ctatccatcc ttgcaaatgt cttctgctga gatgcctcac 180 acggagactg tctctcctct tccctcctcc atggatctgc ttattcagga cagccctgat 240 tcttccacca gtcccaaagg caaacaaccc acttctgcag agaatagtgt cgcaaaaaag 300 gaagacaagg tcccagtcaa gaaacagaag accagaactg tgttctcttc cacccagctg 360 tgtgtactca atgatagatt tcagagacag aaatacctca gcctccagca gatgcaagaa 420 ctctccaaca tcctgaacct cagctacaaa caggtgaaga cctggttcca gaaccagaga 480 atgaaatcta agaggtggca gaaaaacaac tggccgaaga atagcaatgg tgtgacgcag 540 aaggcctcag cacctaccta ccccagcctc tactcttcct accaccaggg atgcctggtg 600 aacccgactg ggaaccttcc aatgtggagc aaccagacct ggaacaattc aacctggagc 660 aaccagaccc agaacatcca gtcctggagc aaccactcct ggaacactca gacctggtgc 720 acccaatcct ggaacaatca ggcctggaac agtcccttct ataactgtgg agaggaatct 780 ctgcagtcct gcatgcagtt ccagccaaat tctcctgcca gtgacttgga ggctgctttg 840 gaagctgctg gggaaggcct taatgtaata cagcagacca ctaggtattt tagtactcca 900 caaaccatgg atttattcct aaactactcc atgaacatgc aacctgaaga cgtgctggcg 960 gtgctggcgg cggcgccgtg a 981 <210> 32 <211> 326 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Nanog-JO-86 MTD Amino Acid Sequence <400> 32 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Ser Val 1 5 10 15 Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe Glu Ala Ser Asp Cys 20 25 30 Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly Pro Glu Glu Asn Tyr 35 40 45 Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro His Thr Glu Thr Val 50 55 60 Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile Gln Asp Ser Pro Asp 65 70 75 80 Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr Ser Ala Glu Asn Ser 85 90 95 Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys Lys Gln Lys Thr Arg 100 105 110 Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu Asn Asp Arg Phe Gln 115 120 125 Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln Glu Leu Ser Asn Ile 130 135 140 Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp Phe Gln Asn Gln Arg 145 150 155 160 Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp Pro Lys Asn Ser Asn 165 170 175 Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr Pro Ser Leu Tyr Ser 180 185 190 Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr Gly Asn Leu Pro Met 195 200 205 Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp Ser Asn Gln Thr Gln 210 215 220 Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn Thr Gln Thr Trp Cys 225 230 235 240 Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser Pro Phe Tyr Asn Cys 245 250 255 Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe Gln Pro Asn Ser Pro 260 265 270 Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala Gly Glu Gly Leu Asn 275 280 285 Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr Pro Gln Thr Met Asp 290 295 300 Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro Glu Asp Val Leu Ala 305 310 315 320 Val Leu Ala Ala Ala Pro 325 <210> 33 <211> 993 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Nanog cDNA Sequence <400> 33 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagag tgtggatcca gcttgtcccc aaagcttgcc ttgctttgaa 120 gcatccgact gtaaagaatc ttcacctatg cctgtgattt gtgggcctga agaaaactat 180 ccatccttgc aaatgtcttc tgctgagatg cctcacacgg agactgtctc tcctcttccc 240 tcctccatgg atctgcttat tcaggacagc cctgattctt ccaccagtcc caaaggcaaa 300 caacccactt ctgcagagaa tagtgtcgca aaaaaggaag acaaggtccc agtcaagaaa 360 cagaagacca gaactgtgtt ctcttccacc cagctgtgtg tactcaatga tagatttcag 420 agacagaaat acctcagcct ccagcagatg caagaactct ccaacatcct gaacctcagc 480 tacaaacagg tgaagacctg gttccagaac cagagaatga aatctaagag gtggcagaaa 540 aacaactggc cgaagaatag caatggtgtg acgcagaagg cctcagcacc tacctacccc 600 agcctctact cttcctacca ccagggatgc ctggtgaacc cgactgggaa ccttccaatg 660 tggagcaacc agacctggaa caattcaacc tggagcaacc agacccagaa catccagtcc 720 tggagcaacc actcctggaa cactcagacc tggtgcaccc aatcctggaa caatcaggcc 780 tggaacagtc ccttctataa ctgtggagag gaatctctgc agtcctgcat gcagttccag 840 ccaaattctc ctgccagtga cttggaggct gctttggaag ctgctgggga aggccttaat 900 gtaatacagc agaccactag gtattttagt actccacaaa ccatggattt attcctaaac 960 tactccatga acatgcaacc tgaagacgtg tga 993 <210> 34 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Nanog Amino Acid Sequence <400> 34 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Ser Val Asp Pro Ala Cys 20 25 30 Pro Gln Ser Leu Pro Cys Phe Glu Ala Ser Asp Cys Lys Glu Ser Ser 35 40 45 Pro Met Pro Val Ile Cys Gly Pro Glu Glu Asn Tyr Pro Ser Leu Gln 50 55 60 Met Ser Ser Ala Glu Met Pro His Thr Glu Thr Val Ser Pro Leu Pro 65 70 75 80 Ser Ser Met Asp Leu Leu Ile Gln Asp Ser Pro Asp Ser Ser Thr Ser 85 90 95 Pro Lys Gly Lys Gln Pro Thr Ser Ala Glu Asn Ser Val Ala Lys Lys 100 105 110 Glu Asp Lys Val Pro Val Lys Lys Gln Lys Thr Arg Thr Val Phe Ser 115 120 125 Ser Thr Gln Leu Cys Val Leu Asn Asp Arg Phe Gln Arg Gln Lys Tyr 130 135 140 Leu Ser Leu Gln Gln Met Gln Glu Leu Ser Asn Ile Leu Asn Leu Ser 145 150 155 160 Tyr Lys Gln Val Lys Thr Trp Phe Gln Asn Gln Arg Met Lys Ser Lys 165 170 175 Arg Trp Gln Lys Asn Asn Trp Pro Lys Asn Ser Asn Gly Val Thr Gln 180 185 190 Lys Ala Ser Ala Pro Thr Tyr Pro Ser Leu Tyr Ser Ser Tyr His Gln 195 200 205 Gly Cys Leu Val Asn Pro Thr Gly Asn Leu Pro Met Trp Ser Asn Gln 210 215 220 Thr Trp Asn Asn Ser Thr Trp Ser Asn Gln Thr Gln Asn Ile Gln Ser 225 230 235 240 Trp Ser Asn His Ser Trp Asn Thr Gln Thr Trp Cys Thr Gln Ser Trp 245 250 255 Asn Asn Gln Ala Trp Asn Ser Pro Phe Tyr Asn Cys Gly Glu Glu Ser 260 265 270 Leu Gln Ser Cys Met Gln Phe Gln Pro Asn Ser Pro Ala Ser Asp Leu 275 280 285 Glu Ala Ala Leu Glu Ala Ala Gly Glu Gly Leu Asn Val Ile Gln Gln 290 295 300 Thr Thr Arg Tyr Phe Ser Thr Pro Gln Thr Met Asp Leu Phe Leu Asn 305 310 315 320 Tyr Ser Met Asn Met Gln Pro Glu Asp Val 325 330 <210> 35 <211> 1020 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Nanog cDNA Sequence <400> 35 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgagtgt ggatccagct 120 tgtccccaaa gcttgccttg ctttgaagca tccgactgta aagaatcttc acctatgcct 180 gtgatttgtg ggcctgaaga aaactatcca tccttgcaaa tgtcttctgc tgagatgcct 240 cacacggaga ctgtctctcc tcttccctcc tccatggatc tgcttattca ggacagccct 300 gattcttcca ccagtcccaa aggcaaacaa cccacttctg cagagaatag tgtcgcaaaa 360 aaggaagaca aggtcccagt caagaaacag aagaccagaa ctgtgttctc ttccacccag 420 ctgtgtgtac tcaatgatag atttcagaga cagaaatacc tcagcctcca gcagatgcaa 480 gaactctcca acatcctgaa cctcagctac aaacaggtga agacctggtt ccagaaccag 540 agaatgaaat ctaagaggtg gcagaaaaac aactggccga agaatagcaa tggtgtgacg 600 cagaaggcct cagcacctac ctaccccagc ctctactctt cctaccacca gggatgcctg 660 gtgaacccga ctgggaacct tccaatgtgg agcaaccaga cctggaacaa ttcaacctgg 720 agcaaccaga cccagaacat ccagtcctgg agcaaccact cctggaacac tcagacctgg 780 tgcacccaat cctggaacaa tcaggcctgg aacagtccct tctataactg tggagaggaa 840 tctctgcagt cctgcatgca gttccagcca aattctcctg ccagtgactt ggaggctgct 900 ttggaagctg ctggggaagg ccttaatgta atacagcaga ccactaggta ttttagtact 960 ccacaaacca tggatttatt cctaaactac tccatgaaca tgcaacctga agacgtgtga 1020 1020 <210> 36 <211> 339 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Nanog Amino Acid Sequence <400> 36 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Ser Val Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe 35 40 45 Glu Ala Ser Asp Cys Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly 50 55 60 Pro Glu Glu Asn Tyr Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro 65 70 75 80 His Thr Glu Thr Val Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile 85 90 95 Gln Asp Ser Pro Asp Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr 100 105 110 Ser Ala Glu Asn Ser Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys 115 120 125 Lys Gln Lys Thr Arg Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu 130 135 140 Asn Asp Arg Phe Gln Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln 145 150 155 160 Glu Leu Ser Asn Ile Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp 165 170 175 Phe Gln Asn Gln Arg Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp 180 185 190 Pro Lys Asn Ser Asn Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr 195 200 205 Pro Ser Leu Tyr Ser Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr 210 215 220 Gly Asn Leu Pro Met Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp 225 230 235 240 Ser Asn Gln Thr Gln Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn 245 250 255 Thr Gln Thr Trp Cys Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser 260 265 270 Pro Phe Tyr Asn Cys Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe 275 280 285 Gln Pro Asn Ser Pro Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala 290 295 300 Gly Glu Gly Leu Asn Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr 305 310 315 320 Pro Gln Thr Met Asp Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro 325 330 335 Glu Asp Val <210> 37 <211> 1020 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Nanog-JO-84 MTD cDNA Sequence <400> 37 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagag tgtggatcca gcttgtcccc aaagcttgcc ttgctttgaa 120 gcatccgact gtaaagaatc ttcacctatg cctgtgattt gtgggcctga agaaaactat 180 ccatccttgc aaatgtcttc tgctgagatg cctcacacgg agactgtctc tcctcttccc 240 tcctccatgg atctgcttat tcaggacagc cctgattctt ccaccagtcc caaaggcaaa 300 caacccactt ctgcagagaa tagtgtcgca aaaaaggaag acaaggtccc agtcaagaaa 360 cagaagacca gaactgtgtt ctcttccacc cagctgtgtg tactcaatga tagatttcag 420 agacagaaat acctcagcct ccagcagatg caagaactct ccaacatcct gaacctcagc 480 tacaaacagg tgaagacctg gttccagaac cagagaatga aatctaagag gtggcagaaa 540 aacaactggc cgaagaatag caatggtgtg acgcagaagg cctcagcacc tacctacccc 600 agcctctact cttcctacca ccagggatgc ctggtgaacc cgactgggaa ccttccaatg 660 tggagcaacc agacctggaa caattcaacc tggagcaacc agacccagaa catccagtcc 720 tggagcaacc actcctggaa cactcagacc tggtgcaccc aatcctggaa caatcaggcc 780 tggaacagtc ccttctataa ctgtggagag gaatctctgc agtcctgcat gcagttccag 840 ccaaattctc ctgccagtga cttggaggct gctttggaag ctgctgggga aggccttaat 900 gtaatacagc agaccactag gtattttagt actccacaaa ccatggattt attcctaaac 960 tactccatga acatgcaacc tgaagacgtg ctggtggcgg cgctgctggc ggtgctgtga 1020 1020 <210> 38 <211> 339 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Nanog-JO-84 MTD Amino Acid Sequence <400> 38 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Ser Val Asp Pro Ala Cys 20 25 30 Pro Gln Ser Leu Pro Cys Phe Glu Ala Ser Asp Cys Lys Glu Ser Ser 35 40 45 Pro Met Pro Val Ile Cys Gly Pro Glu Glu Asn Tyr Pro Ser Leu Gln 50 55 60 Met Ser Ser Ala Glu Met Pro His Thr Glu Thr Val Ser Pro Leu Pro 65 70 75 80 Ser Ser Met Asp Leu Leu Ile Gln Asp Ser Pro Asp Ser Ser Thr Ser 85 90 95 Pro Lys Gly Lys Gln Pro Thr Ser Ala Glu Asn Ser Val Ala Lys Lys 100 105 110 Glu Asp Lys Val Pro Val Lys Lys Gln Lys Thr Arg Thr Val Phe Ser 115 120 125 Ser Thr Gln Leu Cys Val Leu Asn Asp Arg Phe Gln Arg Gln Lys Tyr 130 135 140 Leu Ser Leu Gln Gln Met Gln Glu Leu Ser Asn Ile Leu Asn Leu Ser 145 150 155 160 Tyr Lys Gln Val Lys Thr Trp Phe Gln Asn Gln Arg Met Lys Ser Lys 165 170 175 Arg Trp Gln Lys Asn Asn Trp Pro Lys Asn Ser Asn Gly Val Thr Gln 180 185 190 Lys Ala Ser Ala Pro Thr Tyr Pro Ser Leu Tyr Ser Ser Tyr His Gln 195 200 205 Gly Cys Leu Val Asn Pro Thr Gly Asn Leu Pro Met Trp Ser Asn Gln 210 215 220 Thr Trp Asn Asn Ser Thr Trp Ser Asn Gln Thr Gln Asn Ile Gln Ser 225 230 235 240 Trp Ser Asn His Ser Trp Asn Thr Gln Thr Trp Cys Thr Gln Ser Trp 245 250 255 Asn Asn Gln Ala Trp Asn Ser Pro Phe Tyr Asn Cys Gly Glu Glu Ser 260 265 270 Leu Gln Ser Cys Met Gln Phe Gln Pro Asn Ser Pro Ala Ser Asp Leu 275 280 285 Glu Ala Ala Leu Glu Ala Ala Gly Glu Gly Leu Asn Val Ile Gln Gln 290 295 300 Thr Thr Arg Tyr Phe Ser Thr Pro Gln Thr Met Asp Leu Phe Leu Asn 305 310 315 320 Tyr Ser Met Asn Met Gln Pro Glu Asp Val Leu Val Ala Ala Leu Leu 325 330 335 Ala Val Leu <210> 39 <211> 1047 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Nanog-JO-84 MTD cDNA Sequence <400> 39 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgagtgt ggatccagct 120 tgtccccaaa gcttgccttg ctttgaagca tccgactgta aagaatcttc acctatgcct 180 gtgatttgtg ggcctgaaga aaactatcca tccttgcaaa tgtcttctgc tgagatgcct 240 cacacggaga ctgtctctcc tcttccctcc tccatggatc tgcttattca ggacagccct 300 gattcttcca ccagtcccaa aggcaaacaa cccacttctg cagagaatag tgtcgcaaaa 360 aaggaagaca aggtcccagt caagaaacag aagaccagaa ctgtgttctc ttccacccag 420 ctgtgtgtac tcaatgatag atttcagaga cagaaatacc tcagcctcca gcagatgcaa 480 gaactctcca acatcctgaa cctcagctac aaacaggtga agacctggtt ccagaaccag 540 agaatgaaat ctaagaggtg gcagaaaaac aactggccga agaatagcaa tggtgtgacg 600 cagaaggcct cagcacctac ctaccccagc ctctactctt cctaccacca gggatgcctg 660 gtgaacccga ctgggaacct tccaatgtgg agcaaccaga cctggaacaa ttcaacctgg 720 agcaaccaga cccagaacat ccagtcctgg agcaaccact cctggaacac tcagacctgg 780 tgcacccaat cctggaacaa tcaggcctgg aacagtccct tctataactg tggagaggaa 840 tctctgcagt cctgcatgca gttccagcca aattctcctg ccagtgactt ggaggctgct 900 ttggaagctg ctggggaagg ccttaatgta atacagcaga ccactaggta ttttagtact 960 ccacaaacca tggatttatt cctaaactac tccatgaaca tgcaacctga agacgtgctg 1020 gtggcggcgc tgctggcggt gctgtga 1047 <210> 40 <211> 348 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Nanog-JO-84 MTD Amino Acid Sequence <400> 40 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Ser Val Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe 35 40 45 Glu Ala Ser Asp Cys Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly 50 55 60 Pro Glu Glu Asn Tyr Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro 65 70 75 80 His Thr Glu Thr Val Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile 85 90 95 Gln Asp Ser Pro Asp Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr 100 105 110 Ser Ala Glu Asn Ser Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys 115 120 125 Lys Gln Lys Thr Arg Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu 130 135 140 Asn Asp Arg Phe Gln Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln 145 150 155 160 Glu Leu Ser Asn Ile Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp 165 170 175 Phe Gln Asn Gln Arg Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp 180 185 190 Pro Lys Asn Ser Asn Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr 195 200 205 Pro Ser Leu Tyr Ser Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr 210 215 220 Gly Asn Leu Pro Met Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp 225 230 235 240 Ser Asn Gln Thr Gln Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn 245 250 255 Thr Gln Thr Trp Cys Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser 260 265 270 Pro Phe Tyr Asn Cys Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe 275 280 285 Gln Pro Asn Ser Pro Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala 290 295 300 Gly Glu Gly Leu Asn Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr 305 310 315 320 Pro Gln Thr Met Asp Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro 325 330 335 Glu Asp Val Leu Val Ala Ala Leu Leu Ala Val Leu 340 345 <210> 41 <211> 1017 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Nanog cDNA Sequence <400> 41 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgagtgtgga tccagcttgt 120 ccccaaagct tgccttgctt tgaagcatcc gactgtaaag aatcttcacc tatgcctgtg 180 atttgtgggc ctgaagaaaa ctatccatcc ttgcaaatgt cttctgctga gatgcctcac 240 acggagactg tctctcctct tccctcctcc atggatctgc ttattcagga cagccctgat 300 tcttccacca gtcccaaagg caaacaaccc acttctgcag agaatagtgt cgcaaaaaag 360 gaagacaagg tcccagtcaa gaaacagaag accagaactg tgttctcttc cacccagctg 420 tgtgtactca atgatagatt tcagagacag aaatacctca gcctccagca gatgcaagaa 480 ctctccaaca tcctgaacct cagctacaaa caggtgaaga cctggttcca gaaccagaga 540 atgaaatcta agaggtggca gaaaaacaac tggccgaaga atagcaatgg tgtgacgcag 600 aaggcctcag cacctaccta ccccagcctc tactcttcct accaccaggg atgcctggtg 660 aacccgactg ggaaccttcc aatgtggagc aaccagacct ggaacaattc aacctggagc 720 aaccagaccc agaacatcca gtcctggagc aaccactcct ggaacactca gacctggtgc 780 acccaatcct ggaacaatca ggcctggaac agtcccttct ataactgtgg agaggaatct 840 ctgcagtcct gcatgcagtt ccagccaaat tctcctgcca gtgacttgga ggctgctttg 900 gaagctgctg gggaaggcct taatgtaata cagcagacca ctaggtattt tagtactcca 960 caaaccatgg atttattcct aaactactcc atgaacatgc aacctgaaga cgtgtga 1017 <210> 42 <211> 338 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Nanog Amino Acid Sequence <400> 42 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Ser Val Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe Glu 35 40 45 Ala Ser Asp Cys Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly Pro 50 55 60 Glu Glu Asn Tyr Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro His 65 70 75 80 Thr Glu Thr Val Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile Gln 85 90 95 Asp Ser Pro Asp Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr Ser 100 105 110 Ala Glu Asn Ser Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys Lys 115 120 125 Gln Lys Thr Arg Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu Asn 130 135 140 Asp Arg Phe Gln Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln Glu 145 150 155 160 Leu Ser Asn Ile Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp Phe 165 170 175 Gln Asn Gln Arg Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp Pro 180 185 190 Lys Asn Ser Asn Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr Pro 195 200 205 Ser Leu Tyr Ser Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr Gly 210 215 220 Asn Leu Pro Met Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp Ser 225 230 235 240 Asn Gln Thr Gln Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn Thr 245 250 255 Gln Thr Trp Cys Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser Pro 260 265 270 Phe Tyr Asn Cys Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe Gln 275 280 285 Pro Asn Ser Pro Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala Gly 290 295 300 Glu Gly Leu Asn Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr Pro 305 310 315 320 Gln Thr Met Asp Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro Glu 325 330 335 Asp Val <210> 43 <211> 1017 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Nanog-JO-86 MTD cDNA Sequence <400> 43 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagag tgtggatcca gcttgtcccc aaagcttgcc ttgctttgaa 120 gcatccgact gtaaagaatc ttcacctatg cctgtgattt gtgggcctga agaaaactat 180 ccatccttgc aaatgtcttc tgctgagatg cctcacacgg agactgtctc tcctcttccc 240 tcctccatgg atctgcttat tcaggacagc cctgattctt ccaccagtcc caaaggcaaa 300 caacccactt ctgcagagaa tagtgtcgca aaaaaggaag acaaggtccc agtcaagaaa 360 cagaagacca gaactgtgtt ctcttccacc cagctgtgtg tactcaatga tagatttcag 420 agacagaaat acctcagcct ccagcagatg caagaactct ccaacatcct gaacctcagc 480 tacaaacagg tgaagacctg gttccagaac cagagaatga aatctaagag gtggcagaaa 540 aacaactggc cgaagaatag caatggtgtg acgcagaagg cctcagcacc tacctacccc 600 agcctctact cttcctacca ccagggatgc ctggtgaacc cgactgggaa ccttccaatg 660 tggagcaacc agacctggaa caattcaacc tggagcaacc agacccagaa catccagtcc 720 tggagcaacc actcctggaa cactcagacc tggtgcaccc aatcctggaa caatcaggcc 780 tggaacagtc ccttctataa ctgtggagag gaatctctgc agtcctgcat gcagttccag 840 ccaaattctc ctgccagtga cttggaggct gctttggaag ctgctgggga aggccttaat 900 gtaatacagc agaccactag gtattttagt actccacaaa ccatggattt attcctaaac 960 tactccatga acatgcaacc tgaagacgtg ctggcggtgc tggcggcggc gccgtga 1017 <210> 44 <211> 338 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Nanog-JO-86 MTD Amino Acid Sequence <400> 44 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Ser Val Asp Pro Ala Cys 20 25 30 Pro Gln Ser Leu Pro Cys Phe Glu Ala Ser Asp Cys Lys Glu Ser Ser 35 40 45 Pro Met Pro Val Ile Cys Gly Pro Glu Glu Asn Tyr Pro Ser Leu Gln 50 55 60 Met Ser Ser Ala Glu Met Pro His Thr Glu Thr Val Ser Pro Leu Pro 65 70 75 80 Ser Ser Met Asp Leu Leu Ile Gln Asp Ser Pro Asp Ser Ser Thr Ser 85 90 95 Pro Lys Gly Lys Gln Pro Thr Ser Ala Glu Asn Ser Val Ala Lys Lys 100 105 110 Glu Asp Lys Val Pro Val Lys Lys Gln Lys Thr Arg Thr Val Phe Ser 115 120 125 Ser Thr Gln Leu Cys Val Leu Asn Asp Arg Phe Gln Arg Gln Lys Tyr 130 135 140 Leu Ser Leu Gln Gln Met Gln Glu Leu Ser Asn Ile Leu Asn Leu Ser 145 150 155 160 Tyr Lys Gln Val Lys Thr Trp Phe Gln Asn Gln Arg Met Lys Ser Lys 165 170 175 Arg Trp Gln Lys Asn Asn Trp Pro Lys Asn Ser Asn Gly Val Thr Gln 180 185 190 Lys Ala Ser Ala Pro Thr Tyr Pro Ser Leu Tyr Ser Ser Tyr His Gln 195 200 205 Gly Cys Leu Val Asn Pro Thr Gly Asn Leu Pro Met Trp Ser Asn Gln 210 215 220 Thr Trp Asn Asn Ser Thr Trp Ser Asn Gln Thr Gln Asn Ile Gln Ser 225 230 235 240 Trp Ser Asn His Ser Trp Asn Thr Gln Thr Trp Cys Thr Gln Ser Trp 245 250 255 Asn Asn Gln Ala Trp Asn Ser Pro Phe Tyr Asn Cys Gly Glu Glu Ser 260 265 270 Leu Gln Ser Cys Met Gln Phe Gln Pro Asn Ser Pro Ala Ser Asp Leu 275 280 285 Glu Ala Ala Leu Glu Ala Ala Gly Glu Gly Leu Asn Val Ile Gln Gln 290 295 300 Thr Thr Arg Tyr Phe Ser Thr Pro Gln Thr Met Asp Leu Phe Leu Asn 305 310 315 320 Tyr Ser Met Asn Met Gln Pro Glu Asp Val Leu Ala Val Leu Ala Ala 325 330 335 Ala Pro <210> 45 <211> 1041 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Nanog-JO-86 MTD cDNA Sequence <400> 45 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgagtgtgga tccagcttgt 120 ccccaaagct tgccttgctt tgaagcatcc gactgtaaag aatcttcacc tatgcctgtg 180 atttgtgggc ctgaagaaaa ctatccatcc ttgcaaatgt cttctgctga gatgcctcac 240 acggagactg tctctcctct tccctcctcc atggatctgc ttattcagga cagccctgat 300 tcttccacca gtcccaaagg caaacaaccc acttctgcag agaatagtgt cgcaaaaaag 360 gaagacaagg tcccagtcaa gaaacagaag accagaactg tgttctcttc cacccagctg 420 tgtgtactca atgatagatt tcagagacag aaatacctca gcctccagca gatgcaagaa 480 ctctccaaca tcctgaacct cagctacaaa caggtgaaga cctggttcca gaaccagaga 540 atgaaatcta agaggtggca gaaaaacaac tggccgaaga atagcaatgg tgtgacgcag 600 aaggcctcag cacctaccta ccccagcctc tactcttcct accaccaggg atgcctggtg 660 aacccgactg ggaaccttcc aatgtggagc aaccagacct ggaacaattc aacctggagc 720 aaccagaccc agaacatcca gtcctggagc aaccactcct ggaacactca gacctggtgc 780 acccaatcct ggaacaatca ggcctggaac agtcccttct ataactgtgg agaggaatct 840 ctgcagtcct gcatgcagtt ccagccaaat tctcctgcca gtgacttgga ggctgctttg 900 gaagctgctg gggaaggcct taatgtaata cagcagacca ctaggtattt tagtactcca 960 caaaccatgg atttattcct aaactactcc atgaacatgc aacctgaaga cgtgctggcg 1020 gtgctggcgg cggcgccgtg a 1041 <210> 46 <211> 346 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Nanog-JO-86 MTD Amino Acid Sequence <400> 46 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Ser Val Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe Glu 35 40 45 Ala Ser Asp Cys Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly Pro 50 55 60 Glu Glu Asn Tyr Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro His 65 70 75 80 Thr Glu Thr Val Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile Gln 85 90 95 Asp Ser Pro Asp Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr Ser 100 105 110 Ala Glu Asn Ser Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys Lys 115 120 125 Gln Lys Thr Arg Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu Asn 130 135 140 Asp Arg Phe Gln Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln Glu 145 150 155 160 Leu Ser Asn Ile Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp Phe 165 170 175 Gln Asn Gln Arg Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp Pro 180 185 190 Lys Asn Ser Asn Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr Pro 195 200 205 Ser Leu Tyr Ser Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr Gly 210 215 220 Asn Leu Pro Met Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp Ser 225 230 235 240 Asn Gln Thr Gln Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn Thr 245 250 255 Gln Thr Trp Cys Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser Pro 260 265 270 Phe Tyr Asn Cys Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe Gln 275 280 285 Pro Asn Ser Pro Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala Gly 290 295 300 Glu Gly Leu Asn Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr Pro 305 310 315 320 Gln Thr Met Asp Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro Glu 325 330 335 Asp Val Leu Ala Val Leu Ala Ala Ala Pro 340 345 <210> 47 <211> 1098 <212> DNA <213> Artificial Sequence <220> <223> NLS-Oct4 cDNA Sequence <400> 47 atgaagaaga agaggaaggc gggacacctg gcttcggatt tcgccttctc gccccctcca 60 ggtggtggag gtgatgggcc aggggggccg gagccgggct gggttgatcc tcggacctgg 120 ctaagcttcc aaggccctcc tggagggcca ggaatcgggc cgggggttgg gccaggctct 180 gaggtgtggg ggattccccc atgccccccg ccgtatgagt tctgtggggg gatggcgtac 240 tgtgggcccc aggttggagt ggggctagtg ccccaaggcg gcttggagac ctctcagcct 300 gagggcgaag caggagtcgg ggtggagagc aactccgatg gggcctcccc ggagccctgc 360 accgtcaccc ctggtgccgt gaagctggag aaggagaagc tggagcaaaa cccggaggag 420 tcccaggaca tcaaagctct gcagaaagaa ctcgagcaat ttgccaagct cctgaagcag 480 aagaggatca ccctgggata tacacaggcc gatgtggggc tcaccctggg ggttctattt 540 gggaaggtat tcagccaaac gaccatctgc cgctttgagg ctctgcagct tagcttcaag 600 aacatgtgta agctgcggcc cttgctgcag aagtgggtgg aggaagctga caacaatgaa 660 aatcttcagg agatatgcaa agcagaaacc ctcgtgcagg cccgaaagag aaagcgaacc 720 agtatcgaga accgagtgag aggcaacctg gagaatttgt tcctgcagtg cccgaaaccc 780 acactgcagc agatcagcca catcgcccag cagcttgggc tcgagaagga tgtggtccga 840 gtgtggttct gtaaccggcg ccagaagggc aagcgatcaa gcagcgacta tgcacaacga 900 gaggattttg aggctgctgg gtctcctttc tcagggggac cagtgtcctt tcctctggcc 960 ccagggcccc attttggtac cccaggctat gggagccctc acttcactgc actgtactcc 1020 tcggtccctt tccctgaggg ggaagccttt ccccctgtct ccgtcaccac tctgggctct 1080 cccatgcatt caaactga 1098 <210> 48 <211> 365 <212> PRT <213> Artificial Sequence <220> <223> NLS-Oct4 Amino Acid Sequence <400> 48 Met Lys Lys Lys Arg Lys Ala Gly His Leu Ala Ser Asp Phe Ala Phe 1 5 10 15 Ser Pro Pro Pro Gly Gly Gly Gly Asp Gly Pro Gly Gly Pro Glu Pro 20 25 30 Gly Trp Val Asp Pro Arg Thr Trp Leu Ser Phe Gln Gly Pro Pro Gly 35 40 45 Gly Pro Gly Ile Gly Pro Gly Val Gly Pro Gly Ser Glu Val Trp Gly 50 55 60 Ile Pro Pro Cys Pro Pro Pro Tyr Glu Phe Cys Gly Gly Met Ala Tyr 65 70 75 80 Cys Gly Pro Gln Val Gly Val Gly Leu Val Pro Gln Gly Gly Leu Glu 85 90 95 Thr Ser Gln Pro Glu Gly Glu Ala Gly Val Gly Val Glu Ser Asn Ser 100 105 110 Asp Gly Ala Ser Pro Glu Pro Cys Thr Val Thr Pro Gly Ala Val Lys 115 120 125 Leu Glu Lys Glu Lys Leu Glu Gln Asn Pro Glu Glu Ser Gln Asp Ile 130 135 140 Lys Ala Leu Gln Lys Glu Leu Glu Gln Phe Ala Lys Leu Leu Lys Gln 145 150 155 160 Lys Arg Ile Thr Leu Gly Tyr Thr Gln Ala Asp Val Gly Leu Thr Leu 165 170 175 Gly Val Leu Phe Gly Lys Val Phe Ser Gln Thr Thr Ile Cys Arg Phe 180 185 190 Glu Ala Leu Gln Leu Ser Phe Lys Asn Met Cys Lys Leu Arg Pro Leu 195 200 205 Leu Gln Lys Trp Val Glu Glu Ala Asp Asn Asn Glu Asn Leu Gln Glu 210 215 220 Ile Cys Lys Ala Glu Thr Leu Val Gln Ala Arg Lys Arg Lys Arg Thr 225 230 235 240 Ser Ile Glu Asn Arg Val Arg Gly Asn Leu Glu Asn Leu Phe Leu Gln 245 250 255 Cys Pro Lys Pro Thr Leu Gln Gln Ile Ser His Ile Ala Gln Gln Leu 260 265 270 Gly Leu Glu Lys Asp Val Val Arg Val Trp Phe Cys Asn Arg Arg Gln 275 280 285 Lys Gly Lys Arg Ser Ser Ser Asp Tyr Ala Gln Arg Glu Asp Phe Glu 290 295 300 Ala Ala Gly Ser Pro Phe Ser Gly Gly Pro Val Ser Phe Pro Leu Ala 305 310 315 320 Pro Gly Pro His Phe Gly Thr Pro Gly Tyr Gly Ser Pro His Phe Thr 325 330 335 Ala Leu Tyr Ser Ser Val Pro Phe Pro Glu Gly Glu Ala Phe Pro Pro 340 345 350 Val Ser Val Thr Thr Leu Gly Ser Pro Met His Ser Asn 355 360 365 <210> 49 <211> 1125 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Oct4 cDNA Sequence <400> 49 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctggcggg acacctggct 60 tcggatttcg ccttctcgcc ccctccaggt ggtggaggtg atgggccagg ggggccggag 120 ccgggctggg ttgatcctcg gacctggcta agcttccaag gccctcctgg agggccagga 180 atcgggccgg gggttgggcc aggctctgag gtgtggggga ttcccccatg ccccccgccg 240 tatgagttct gtggggggat ggcgtactgt gggccccagg ttggagtggg gctagtgccc 300 caaggcggct tggagacctc tcagcctgag ggcgaagcag gagtcggggt ggagagcaac 360 tccgatgggg cctccccgga gccctgcacc gtcacccctg gtgccgtgaa gctggagaag 420 gagaagctgg agcaaaaccc ggaggagtcc caggacatca aagctctgca gaaagaactc 480 gagcaatttg ccaagctcct gaagcagaag aggatcaccc tgggatatac acaggccgat 540 gtggggctca ccctgggggt tctatttggg aaggtattca gccaaacgac catctgccgc 600 tttgaggctc tgcagcttag cttcaagaac atgtgtaagc tgcggccctt gctgcagaag 660 tgggtggagg aagctgacaa caatgaaaat cttcaggaga tatgcaaagc agaaaccctc 720 gtgcaggccc gaaagagaaa gcgaaccagt atcgagaacc gagtgagagg caacctggag 780 aatttgttcc tgcagtgccc gaaacccaca ctgcagcaga tcagccacat cgcccagcag 840 cttgggctcg agaaggatgt ggtccgagtg tggttctgta accggcgcca gaagggcaag 900 cgatcaagca gcgactatgc acaacgagag gattttgagg ctgctgggtc tcctttctca 960 gggggaccag tgtcctttcc tctggcccca gggccccatt ttggtacccc aggctatggg 1020 agccctcact tcactgcact gtactcctcg gtccctttcc ctgaggggga agcctttccc 1080 cctgtctccg tcaccactct gggctctccc atgcattcaa actga 1125 <210> 50 <211> 374 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Oct4 Amino Acid Sequence <400> 50 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Ala 1 5 10 15 Gly His Leu Ala Ser Asp Phe Ala Phe Ser Pro Pro Pro Gly Gly Gly 20 25 30 Gly Asp Gly Pro Gly Gly Pro Glu Pro Gly Trp Val Asp Pro Arg Thr 35 40 45 Trp Leu Ser Phe Gln Gly Pro Pro Gly Gly Pro Gly Ile Gly Pro Gly 50 55 60 Val Gly Pro Gly Ser Glu Val Trp Gly Ile Pro Pro Cys Pro Pro Pro 65 70 75 80 Tyr Glu Phe Cys Gly Gly Met Ala Tyr Cys Gly Pro Gln Val Gly Val 85 90 95 Gly Leu Val Pro Gln Gly Gly Leu Glu Thr Ser Gln Pro Glu Gly Glu 100 105 110 Ala Gly Val Gly Val Glu Ser Asn Ser Asp Gly Ala Ser Pro Glu Pro 115 120 125 Cys Thr Val Thr Pro Gly Ala Val Lys Leu Glu Lys Glu Lys Leu Glu 130 135 140 Gln Asn Pro Glu Glu Ser Gln Asp Ile Lys Ala Leu Gln Lys Glu Leu 145 150 155 160 Glu Gln Phe Ala Lys Leu Leu Lys Gln Lys Arg Ile Thr Leu Gly Tyr 165 170 175 Thr Gln Ala Asp Val Gly Leu Thr Leu Gly Val Leu Phe Gly Lys Val 180 185 190 Phe Ser Gln Thr Thr Ile Cys Arg Phe Glu Ala Leu Gln Leu Ser Phe 195 200 205 Lys Asn Met Cys Lys Leu Arg Pro Leu Leu Gln Lys Trp Val Glu Glu 210 215 220 Ala Asp Asn Asn Glu Asn Leu Gln Glu Ile Cys Lys Ala Glu Thr Leu 225 230 235 240 Val Gln Ala Arg Lys Arg Lys Arg Thr Ser Ile Glu Asn Arg Val Arg 245 250 255 Gly Asn Leu Glu Asn Leu Phe Leu Gln Cys Pro Lys Pro Thr Leu Gln 260 265 270 Gln Ile Ser His Ile Ala Gln Gln Leu Gly Leu Glu Lys Asp Val Val 275 280 285 Arg Val Trp Phe Cys Asn Arg Arg Gln Lys Gly Lys Arg Ser Ser Ser 290 295 300 Asp Tyr Ala Gln Arg Glu Asp Phe Glu Ala Ala Gly Ser Pro Phe Ser 305 310 315 320 Gly Gly Pro Val Ser Phe Pro Leu Ala Pro Gly Pro His Phe Gly Thr 325 330 335 Pro Gly Tyr Gly Ser Pro His Phe Thr Ala Leu Tyr Ser Ser Val Pro 340 345 350 Phe Pro Glu Gly Glu Ala Phe Pro Pro Val Ser Val Thr Thr Leu Gly 355 360 365 Ser Pro Met His Ser Asn 370 <210> 51 <211> 1125 <212> DNA <213> Artificial Sequence <220> <223> NLS-Oct4-JO-84 MTD cDNA Sequence <400> 51 atgaagaaga agaggaaggc gggacacctg gcttcggatt tcgccttctc gccccctcca 60 ggtggtggag gtgatgggcc aggggggccg gagccgggct gggttgatcc tcggacctgg 120 ctaagcttcc aaggccctcc tggagggcca ggaatcgggc cgggggttgg gccaggctct 180 gaggtgtggg ggattccccc atgccccccg ccgtatgagt tctgtggggg gatggcgtac 240 tgtgggcccc aggttggagt ggggctagtg ccccaaggcg gcttggagac ctctcagcct 300 gagggcgaag caggagtcgg ggtggagagc aactccgatg gggcctcccc ggagccctgc 360 accgtcaccc ctggtgccgt gaagctggag aaggagaagc tggagcaaaa cccggaggag 420 tcccaggaca tcaaagctct gcagaaagaa ctcgagcaat ttgccaagct cctgaagcag 480 aagaggatca ccctgggata tacacaggcc gatgtggggc tcaccctggg ggttctattt 540 gggaaggtat tcagccaaac gaccatctgc cgctttgagg ctctgcagct tagcttcaag 600 aacatgtgta agctgcggcc cttgctgcag aagtgggtgg aggaagctga caacaatgaa 660 aatcttcagg agatatgcaa agcagaaacc ctcgtgcagg cccgaaagag aaagcgaacc 720 agtatcgaga accgagtgag aggcaacctg gagaatttgt tcctgcagtg cccgaaaccc 780 acactgcagc agatcagcca catcgcccag cagcttgggc tcgagaagga tgtggtccga 840 gtgtggttct gtaaccggcg ccagaagggc aagcgatcaa gcagcgacta tgcacaacga 900 gaggattttg aggctgctgg gtctcctttc tcagggggac cagtgtcctt tcctctggcc 960 ccagggcccc attttggtac cccaggctat gggagccctc acttcactgc actgtactcc 1020 tcggtccctt tccctgaggg ggaagccttt ccccctgtct ccgtcaccac tctgggctct 1080 cccatgcatt caaacctggt ggcggcgctg ctggcggtgc tgtga 1125 <210> 52 <211> 374 <212> PRT <213> Artificial Sequence <220> <223> NLS-Oct4-JO-84 MTD Amino Acid Sequence <400> 52 Met Lys Lys Lys Arg Lys Ala Gly His Leu Ala Ser Asp Phe Ala Phe 1 5 10 15 Ser Pro Pro Pro Gly Gly Gly Gly Asp Gly Pro Gly Gly Pro Glu Pro 20 25 30 Gly Trp Val Asp Pro Arg Thr Trp Leu Ser Phe Gln Gly Pro Pro Gly 35 40 45 Gly Pro Gly Ile Gly Pro Gly Val Gly Pro Gly Ser Glu Val Trp Gly 50 55 60 Ile Pro Pro Cys Pro Pro Pro Tyr Glu Phe Cys Gly Gly Met Ala Tyr 65 70 75 80 Cys Gly Pro Gln Val Gly Val Gly Leu Val Pro Gln Gly Gly Leu Glu 85 90 95 Thr Ser Gln Pro Glu Gly Glu Ala Gly Val Gly Val Glu Ser Asn Ser 100 105 110 Asp Gly Ala Ser Pro Glu Pro Cys Thr Val Thr Pro Gly Ala Val Lys 115 120 125 Leu Glu Lys Glu Lys Leu Glu Gln Asn Pro Glu Glu Ser Gln Asp Ile 130 135 140 Lys Ala Leu Gln Lys Glu Leu Glu Gln Phe Ala Lys Leu Leu Lys Gln 145 150 155 160 Lys Arg Ile Thr Leu Gly Tyr Thr Gln Ala Asp Val Gly Leu Thr Leu 165 170 175 Gly Val Leu Phe Gly Lys Val Phe Ser Gln Thr Thr Ile Cys Arg Phe 180 185 190 Glu Ala Leu Gln Leu Ser Phe Lys Asn Met Cys Lys Leu Arg Pro Leu 195 200 205 Leu Gln Lys Trp Val Glu Glu Ala Asp Asn Asn Glu Asn Leu Gln Glu 210 215 220 Ile Cys Lys Ala Glu Thr Leu Val Gln Ala Arg Lys Arg Lys Arg Thr 225 230 235 240 Ser Ile Glu Asn Arg Val Arg Gly Asn Leu Glu Asn Leu Phe Leu Gln 245 250 255 Cys Pro Lys Pro Thr Leu Gln Gln Ile Ser His Ile Ala Gln Gln Leu 260 265 270 Gly Leu Glu Lys Asp Val Val Arg Val Trp Phe Cys Asn Arg Arg Gln 275 280 285 Lys Gly Lys Arg Ser Ser Ser Asp Tyr Ala Gln Arg Glu Asp Phe Glu 290 295 300 Ala Ala Gly Ser Pro Phe Ser Gly Gly Pro Val Ser Phe Pro Leu Ala 305 310 315 320 Pro Gly Pro His Phe Gly Thr Pro Gly Tyr Gly Ser Pro His Phe Thr 325 330 335 Ala Leu Tyr Ser Ser Val Pro Phe Pro Glu Gly Glu Ala Phe Pro Pro 340 345 350 Val Ser Val Thr Thr Leu Gly Ser Pro Met His Ser Asn Leu Val Ala 355 360 365 Ala Leu Leu Ala Val Leu 370 <210> 53 <211> 1152 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Oct4-JO-84 MTD cDNA Sequence <400> 53 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctggcggg acacctggct 60 tcggatttcg ccttctcgcc ccctccaggt ggtggaggtg atgggccagg ggggccggag 120 ccgggctggg ttgatcctcg gacctggcta agcttccaag gccctcctgg agggccagga 180 atcgggccgg gggttgggcc aggctctgag gtgtggggga ttcccccatg ccccccgccg 240 tatgagttct gtggggggat ggcgtactgt gggccccagg ttggagtggg gctagtgccc 300 caaggcggct tggagacctc tcagcctgag ggcgaagcag gagtcggggt ggagagcaac 360 tccgatgggg cctccccgga gccctgcacc gtcacccctg gtgccgtgaa gctggagaag 420 gagaagctgg agcaaaaccc ggaggagtcc caggacatca aagctctgca gaaagaactc 480 gagcaatttg ccaagctcct gaagcagaag aggatcaccc tgggatatac acaggccgat 540 gtggggctca ccctgggggt tctatttggg aaggtattca gccaaacgac catctgccgc 600 tttgaggctc tgcagcttag cttcaagaac atgtgtaagc tgcggccctt gctgcagaag 660 tgggtggagg aagctgacaa caatgaaaat cttcaggaga tatgcaaagc agaaaccctc 720 gtgcaggccc gaaagagaaa gcgaaccagt atcgagaacc gagtgagagg caacctggag 780 aatttgttcc tgcagtgccc gaaacccaca ctgcagcaga tcagccacat cgcccagcag 840 cttgggctcg agaaggatgt ggtccgagtg tggttctgta accggcgcca gaagggcaag 900 cgatcaagca gcgactatgc acaacgagag gattttgagg ctgctgggtc tcctttctca 960 gggggaccag tgtcctttcc tctggcccca gggccccatt ttggtacccc aggctatggg 1020 agccctcact tcactgcact gtactcctcg gtccctttcc ctgaggggga agcctttccc 1080 cctgtctccg tcaccactct gggctctccc atgcattcaa acctggtggc ggcgctgctg 1140 gcggtgctgt ga 1152 <210> 54 <211> 383 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Oct4-JO-84 MTD Amino Acid Sequence <400> 54 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Ala 1 5 10 15 Gly His Leu Ala Ser Asp Phe Ala Phe Ser Pro Pro Pro Gly Gly Gly 20 25 30 Gly Asp Gly Pro Gly Gly Pro Glu Pro Gly Trp Val Asp Pro Arg Thr 35 40 45 Trp Leu Ser Phe Gln Gly Pro Pro Gly Gly Pro Gly Ile Gly Pro Gly 50 55 60 Val Gly Pro Gly Ser Glu Val Trp Gly Ile Pro Pro Cys Pro Pro Pro 65 70 75 80 Tyr Glu Phe Cys Gly Gly Met Ala Tyr Cys Gly Pro Gln Val Gly Val 85 90 95 Gly Leu Val Pro Gln Gly Gly Leu Glu Thr Ser Gln Pro Glu Gly Glu 100 105 110 Ala Gly Val Gly Val Glu Ser Asn Ser Asp Gly Ala Ser Pro Glu Pro 115 120 125 Cys Thr Val Thr Pro Gly Ala Val Lys Leu Glu Lys Glu Lys Leu Glu 130 135 140 Gln Asn Pro Glu Glu Ser Gln Asp Ile Lys Ala Leu Gln Lys Glu Leu 145 150 155 160 Glu Gln Phe Ala Lys Leu Leu Lys Gln Lys Arg Ile Thr Leu Gly Tyr 165 170 175 Thr Gln Ala Asp Val Gly Leu Thr Leu Gly Val Leu Phe Gly Lys Val 180 185 190 Phe Ser Gln Thr Thr Ile Cys Arg Phe Glu Ala Leu Gln Leu Ser Phe 195 200 205 Lys Asn Met Cys Lys Leu Arg Pro Leu Leu Gln Lys Trp Val Glu Glu 210 215 220 Ala Asp Asn Asn Glu Asn Leu Gln Glu Ile Cys Lys Ala Glu Thr Leu 225 230 235 240 Val Gln Ala Arg Lys Arg Lys Arg Thr Ser Ile Glu Asn Arg Val Arg 245 250 255 Gly Asn Leu Glu Asn Leu Phe Leu Gln Cys Pro Lys Pro Thr Leu Gln 260 265 270 Gln Ile Ser His Ile Ala Gln Gln Leu Gly Leu Glu Lys Asp Val Val 275 280 285 Arg Val Trp Phe Cys Asn Arg Arg Gln Lys Gly Lys Arg Ser Ser Ser 290 295 300 Asp Tyr Ala Gln Arg Glu Asp Phe Glu Ala Ala Gly Ser Pro Phe Ser 305 310 315 320 Gly Gly Pro Val Ser Phe Pro Leu Ala Pro Gly Pro His Phe Gly Thr 325 330 335 Pro Gly Tyr Gly Ser Pro His Phe Thr Ala Leu Tyr Ser Ser Val Pro 340 345 350 Phe Pro Glu Gly Glu Ala Phe Pro Pro Val Ser Val Thr Thr Leu Gly 355 360 365 Ser Pro Met His Ser Asn Leu Val Ala Ala Leu Leu Ala Val Leu 370 375 380 <210> 55 <211> 1122 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Oct4 cDNA Sequence <400> 55 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cggcgggaca cctggcttcg 60 gatttcgcct tctcgccccc tccaggtggt ggaggtgatg ggccaggggg gccggagccg 120 ggctgggttg atcctcggac ctggctaagc ttccaaggcc ctcctggagg gccaggaatc 180 gggccggggg ttgggccagg ctctgaggtg tgggggattc ccccatgccc cccgccgtat 240 gagttctgtg gggggatggc gtactgtggg ccccaggttg gagtggggct agtgccccaa 300 ggcggcttgg agacctctca gcctgagggc gaagcaggag tcggggtgga gagcaactcc 360 gatggggcct ccccggagcc ctgcaccgtc acccctggtg ccgtgaagct ggagaaggag 420 aagctggagc aaaacccgga ggagtcccag gacatcaaag ctctgcagaa agaactcgag 480 caatttgcca agctcctgaa gcagaagagg atcaccctgg gatatacaca ggccgatgtg 540 gggctcaccc tgggggttct atttgggaag gtattcagcc aaacgaccat ctgccgcttt 600 gaggctctgc agcttagctt caagaacatg tgtaagctgc ggcccttgct gcagaagtgg 660 gtggaggaag ctgacaacaa tgaaaatctt caggagatat gcaaagcaga aaccctcgtg 720 caggcccgaa agagaaagcg aaccagtatc gagaaccgag tgagaggcaa cctggagaat 780 ttgttcctgc agtgcccgaa acccacactg cagcagatca gccacatcgc ccagcagctt 840 gggctcgaga aggatgtggt ccgagtgtgg ttctgtaacc ggcgccagaa gggcaagcga 900 tcaagcagcg actatgcaca acgagaggat tttgaggctg ctgggtctcc tttctcaggg 960 ggaccagtgt cctttcctct ggccccaggg ccccattttg gtaccccagg ctatgggagc 1020 cctcacttca ctgcactgta ctcctcggtc cctttccctg agggggaagc ctttccccct 1080 gtctccgtca ccactctggg ctctcccatg cattcaaact ga 1122 <210> 56 <211> 373 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Oct4 Amico Acid Sequence <400> 56 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Ala Gly 1 5 10 15 His Leu Ala Ser Asp Phe Ala Phe Ser Pro Pro Pro Gly Gly Gly Gly 20 25 30 Asp Gly Pro Gly Gly Pro Glu Pro Gly Trp Val Asp Pro Arg Thr Trp 35 40 45 Leu Ser Phe Gln Gly Pro Pro Gly Gly Pro Gly Ile Gly Pro Gly Val 50 55 60 Gly Pro Gly Ser Glu Val Trp Gly Ile Pro Pro Cys Pro Pro Pro Tyr 65 70 75 80 Glu Phe Cys Gly Gly Met Ala Tyr Cys Gly Pro Gln Val Gly Val Gly 85 90 95 Leu Val Pro Gln Gly Gly Leu Glu Thr Ser Gln Pro Glu Gly Glu Ala 100 105 110 Gly Val Gly Val Glu Ser Asn Ser Asp Gly Ala Ser Pro Glu Pro Cys 115 120 125 Thr Val Thr Pro Gly Ala Val Lys Leu Glu Lys Glu Lys Leu Glu Gln 130 135 140 Asn Pro Glu Glu Ser Gln Asp Ile Lys Ala Leu Gln Lys Glu Leu Glu 145 150 155 160 Gln Phe Ala Lys Leu Leu Lys Gln Lys Arg Ile Thr Leu Gly Tyr Thr 165 170 175 Gln Ala Asp Val Gly Leu Thr Leu Gly Val Leu Phe Gly Lys Val Phe 180 185 190 Ser Gln Thr Thr Ile Cys Arg Phe Glu Ala Leu Gln Leu Ser Phe Lys 195 200 205 Asn Met Cys Lys Leu Arg Pro Leu Leu Gln Lys Trp Val Glu Glu Ala 210 215 220 Asp Asn Asn Glu Asn Leu Gln Glu Ile Cys Lys Ala Glu Thr Leu Val 225 230 235 240 Gln Ala Arg Lys Arg Lys Arg Thr Ser Ile Glu Asn Arg Val Arg Gly 245 250 255 Asn Leu Glu Asn Leu Phe Leu Gln Cys Pro Lys Pro Thr Leu Gln Gln 260 265 270 Ile Ser His Ile Ala Gln Gln Leu Gly Leu Glu Lys Asp Val Val Arg 275 280 285 Val Trp Phe Cys Asn Arg Arg Gln Lys Gly Lys Arg Ser Ser Ser Asp 290 295 300 Tyr Ala Gln Arg Glu Asp Phe Glu Ala Ala Gly Ser Pro Phe Ser Gly 305 310 315 320 Gly Pro Val Ser Phe Pro Leu Ala Pro Gly Pro His Phe Gly Thr Pro 325 330 335 Gly Tyr Gly Ser Pro His Phe Thr Ala Leu Tyr Ser Ser Val Pro Phe 340 345 350 Pro Glu Gly Glu Ala Phe Pro Pro Val Ser Val Thr Thr Leu Gly Ser 355 360 365 Pro Met His Ser Asn 370 <210> 57 <211> 1122 <212> DNA <213> Artificial Sequence <220> <223> NLS-Oct4-JO-86 MTD cDNA Sequence <400> 57 atgaagaaga agaggaaggc gggacacctg gcttcggatt tcgccttctc gccccctcca 60 ggtggtggag gtgatgggcc aggggggccg gagccgggct gggttgatcc tcggacctgg 120 ctaagcttcc aaggccctcc tggagggcca ggaatcgggc cgggggttgg gccaggctct 180 gaggtgtggg ggattccccc atgccccccg ccgtatgagt tctgtggggg gatggcgtac 240 tgtgggcccc aggttggagt ggggctagtg ccccaaggcg gcttggagac ctctcagcct 300 gagggcgaag caggagtcgg ggtggagagc aactccgatg gggcctcccc ggagccctgc 360 accgtcaccc ctggtgccgt gaagctggag aaggagaagc tggagcaaaa cccggaggag 420 tcccaggaca tcaaagctct gcagaaagaa ctcgagcaat ttgccaagct cctgaagcag 480 aagaggatca ccctgggata tacacaggcc gatgtggggc tcaccctggg ggttctattt 540 gggaaggtat tcagccaaac gaccatctgc cgctttgagg ctctgcagct tagcttcaag 600 aacatgtgta agctgcggcc cttgctgcag aagtgggtgg aggaagctga caacaatgaa 660 aatcttcagg agatatgcaa agcagaaacc ctcgtgcagg cccgaaagag aaagcgaacc 720 agtatcgaga accgagtgag aggcaacctg gagaatttgt tcctgcagtg cccgaaaccc 780 acactgcagc agatcagcca catcgcccag cagcttgggc tcgagaagga tgtggtccga 840 gtgtggttct gtaaccggcg ccagaagggc aagcgatcaa gcagcgacta tgcacaacga 900 gaggattttg aggctgctgg gtctcctttc tcagggggac cagtgtcctt tcctctggcc 960 ccagggcccc attttggtac cccaggctat gggagccctc acttcactgc actgtactcc 1020 tcggtccctt tccctgaggg ggaagccttt ccccctgtct ccgtcaccac tctgggctct 1080 cccatgcatt caaacctggc ggtgctggcg gcggcgccgt ga 1122 <210> 58 <211> 373 <212> PRT <213> Artificial Sequence <220> <223> NLS-Oct4-JO-86 MTD Amino Acid Sequence <400> 58 Met Lys Lys Lys Arg Lys Ala Gly His Leu Ala Ser Asp Phe Ala Phe 1 5 10 15 Ser Pro Pro Pro Gly Gly Gly Gly Asp Gly Pro Gly Gly Pro Glu Pro 20 25 30 Gly Trp Val Asp Pro Arg Thr Trp Leu Ser Phe Gln Gly Pro Pro Gly 35 40 45 Gly Pro Gly Ile Gly Pro Gly Val Gly Pro Gly Ser Glu Val Trp Gly 50 55 60 Ile Pro Pro Cys Pro Pro Pro Tyr Glu Phe Cys Gly Gly Met Ala Tyr 65 70 75 80 Cys Gly Pro Gln Val Gly Val Gly Leu Val Pro Gln Gly Gly Leu Glu 85 90 95 Thr Ser Gln Pro Glu Gly Glu Ala Gly Val Gly Val Glu Ser Asn Ser 100 105 110 Asp Gly Ala Ser Pro Glu Pro Cys Thr Val Thr Pro Gly Ala Val Lys 115 120 125 Leu Glu Lys Glu Lys Leu Glu Gln Asn Pro Glu Glu Ser Gln Asp Ile 130 135 140 Lys Ala Leu Gln Lys Glu Leu Glu Gln Phe Ala Lys Leu Leu Lys Gln 145 150 155 160 Lys Arg Ile Thr Leu Gly Tyr Thr Gln Ala Asp Val Gly Leu Thr Leu 165 170 175 Gly Val Leu Phe Gly Lys Val Phe Ser Gln Thr Thr Ile Cys Arg Phe 180 185 190 Glu Ala Leu Gln Leu Ser Phe Lys Asn Met Cys Lys Leu Arg Pro Leu 195 200 205 Leu Gln Lys Trp Val Glu Glu Ala Asp Asn Asn Glu Asn Leu Gln Glu 210 215 220 Ile Cys Lys Ala Glu Thr Leu Val Gln Ala Arg Lys Arg Lys Arg Thr 225 230 235 240 Ser Ile Glu Asn Arg Val Arg Gly Asn Leu Glu Asn Leu Phe Leu Gln 245 250 255 Cys Pro Lys Pro Thr Leu Gln Gln Ile Ser His Ile Ala Gln Gln Leu 260 265 270 Gly Leu Glu Lys Asp Val Val Arg Val Trp Phe Cys Asn Arg Arg Gln 275 280 285 Lys Gly Lys Arg Ser Ser Ser Asp Tyr Ala Gln Arg Glu Asp Phe Glu 290 295 300 Ala Ala Gly Ser Pro Phe Ser Gly Gly Pro Val Ser Phe Pro Leu Ala 305 310 315 320 Pro Gly Pro His Phe Gly Thr Pro Gly Tyr Gly Ser Pro His Phe Thr 325 330 335 Ala Leu Tyr Ser Ser Val Pro Phe Pro Glu Gly Glu Ala Phe Pro Pro 340 345 350 Val Ser Val Thr Thr Leu Gly Ser Pro Met His Ser Asn Leu Ala Val 355 360 365 Leu Ala Ala Ala Pro 370 <210> 59 <211> 1146 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Oct4-JO-86 MTD cDNA Sequence <400> 59 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cggcgggaca cctggcttcg 60 gatttcgcct tctcgccccc tccaggtggt ggaggtgatg ggccaggggg gccggagccg 120 ggctgggttg atcctcggac ctggctaagc ttccaaggcc ctcctggagg gccaggaatc 180 gggccggggg ttgggccagg ctctgaggtg tgggggattc ccccatgccc cccgccgtat 240 gagttctgtg gggggatggc gtactgtggg ccccaggttg gagtggggct agtgccccaa 300 ggcggcttgg agacctctca gcctgagggc gaagcaggag tcggggtgga gagcaactcc 360 gatggggcct ccccggagcc ctgcaccgtc acccctggtg ccgtgaagct ggagaaggag 420 aagctggagc aaaacccgga ggagtcccag gacatcaaag ctctgcagaa agaactcgag 480 caatttgcca agctcctgaa gcagaagagg atcaccctgg gatatacaca ggccgatgtg 540 gggctcaccc tgggggttct atttgggaag gtattcagcc aaacgaccat ctgccgcttt 600 gaggctctgc agcttagctt caagaacatg tgtaagctgc ggcccttgct gcagaagtgg 660 gtggaggaag ctgacaacaa tgaaaatctt caggagatat gcaaagcaga aaccctcgtg 720 caggcccgaa agagaaagcg aaccagtatc gagaaccgag tgagaggcaa cctggagaat 780 ttgttcctgc agtgcccgaa acccacactg cagcagatca gccacatcgc ccagcagctt 840 gggctcgaga aggatgtggt ccgagtgtgg ttctgtaacc ggcgccagaa gggcaagcga 900 tcaagcagcg actatgcaca acgagaggat tttgaggctg ctgggtctcc tttctcaggg 960 ggaccagtgt cctttcctct ggccccaggg ccccattttg gtaccccagg ctatgggagc 1020 cctcacttca ctgcactgta ctcctcggtc cctttccctg agggggaagc ctttccccct 1080 gtctccgtca ccactctggg ctctcccatg cattcaaacc tggcggtgct ggcggcggcg 1140 ccgtga 1146 <210> 60 <211> 381 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Oct4-JO-86 MTD Amino Acid Sequence <400> 60 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Ala Gly 1 5 10 15 His Leu Ala Ser Asp Phe Ala Phe Ser Pro Pro Pro Gly Gly Gly Gly 20 25 30 Asp Gly Pro Gly Gly Pro Glu Pro Gly Trp Val Asp Pro Arg Thr Trp 35 40 45 Leu Ser Phe Gln Gly Pro Pro Gly Gly Pro Gly Ile Gly Pro Gly Val 50 55 60 Gly Pro Gly Ser Glu Val Trp Gly Ile Pro Pro Cys Pro Pro Pro Tyr 65 70 75 80 Glu Phe Cys Gly Gly Met Ala Tyr Cys Gly Pro Gln Val Gly Val Gly 85 90 95 Leu Val Pro Gln Gly Gly Leu Glu Thr Ser Gln Pro Glu Gly Glu Ala 100 105 110 Gly Val Gly Val Glu Ser Asn Ser Asp Gly Ala Ser Pro Glu Pro Cys 115 120 125 Thr Val Thr Pro Gly Ala Val Lys Leu Glu Lys Glu Lys Leu Glu Gln 130 135 140 Asn Pro Glu Glu Ser Gln Asp Ile Lys Ala Leu Gln Lys Glu Leu Glu 145 150 155 160 Gln Phe Ala Lys Leu Leu Lys Gln Lys Arg Ile Thr Leu Gly Tyr Thr 165 170 175 Gln Ala Asp Val Gly Leu Thr Leu Gly Val Leu Phe Gly Lys Val Phe 180 185 190 Ser Gln Thr Thr Ile Cys Arg Phe Glu Ala Leu Gln Leu Ser Phe Lys 195 200 205 Asn Met Cys Lys Leu Arg Pro Leu Leu Gln Lys Trp Val Glu Glu Ala 210 215 220 Asp Asn Asn Glu Asn Leu Gln Glu Ile Cys Lys Ala Glu Thr Leu Val 225 230 235 240 Gln Ala Arg Lys Arg Lys Arg Thr Ser Ile Glu Asn Arg Val Arg Gly 245 250 255 Asn Leu Glu Asn Leu Phe Leu Gln Cys Pro Lys Pro Thr Leu Gln Gln 260 265 270 Ile Ser His Ile Ala Gln Gln Leu Gly Leu Glu Lys Asp Val Val Arg 275 280 285 Val Trp Phe Cys Asn Arg Arg Gln Lys Gly Lys Arg Ser Ser Ser Asp 290 295 300 Tyr Ala Gln Arg Glu Asp Phe Glu Ala Ala Gly Ser Pro Phe Ser Gly 305 310 315 320 Gly Pro Val Ser Phe Pro Leu Ala Pro Gly Pro His Phe Gly Thr Pro 325 330 335 Gly Tyr Gly Ser Pro His Phe Thr Ala Leu Tyr Ser Ser Val Pro Phe 340 345 350 Pro Glu Gly Glu Ala Phe Pro Pro Val Ser Val Thr Thr Leu Gly Ser 355 360 365 Pro Met His Ser Asn Leu Ala Val Leu Ala Ala Ala Pro 370 375 380 <210> 61 <211> 1158 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Oct4 cDNA Sequence <400> 61 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaaggc gggacacctg gcttcggatt tcgccttctc gccccctcca 120 ggtggtggag gtgatgggcc aggggggccg gagccgggct gggttgatcc tcggacctgg 180 ctaagcttcc aaggccctcc tggagggcca ggaatcgggc cgggggttgg gccaggctct 240 gaggtgtggg ggattccccc atgccccccg ccgtatgagt tctgtggggg gatggcgtac 300 tgtgggcccc aggttggagt ggggctagtg ccccaaggcg gcttggagac ctctcagcct 360 gagggcgaag caggagtcgg ggtggagagc aactccgatg gggcctcccc ggagccctgc 420 accgtcaccc ctggtgccgt gaagctggag aaggagaagc tggagcaaaa cccggaggag 480 tcccaggaca tcaaagctct gcagaaagaa ctcgagcaat ttgccaagct cctgaagcag 540 aagaggatca ccctgggata tacacaggcc gatgtggggc tcaccctggg ggttctattt 600 gggaaggtat tcagccaaac gaccatctgc cgctttgagg ctctgcagct tagcttcaag 660 aacatgtgta agctgcggcc cttgctgcag aagtgggtgg aggaagctga caacaatgaa 720 aatcttcagg agatatgcaa agcagaaacc ctcgtgcagg cccgaaagag aaagcgaacc 780 agtatcgaga accgagtgag aggcaacctg gagaatttgt tcctgcagtg cccgaaaccc 840 acactgcagc agatcagcca catcgcccag cagcttgggc tcgagaagga tgtggtccga 900 gtgtggttct gtaaccggcg ccagaagggc aagcgatcaa gcagcgacta tgcacaacga 960 gaggattttg aggctgctgg gtctcctttc tcagggggac cagtgtcctt tcctctggcc 1020 ccagggcccc attttggtac cccaggctat gggagccctc acttcactgc actgtactcc 1080 tcggtccctt tccctgaggg ggaagccttt ccccctgtct ccgtcaccac tctgggctct 1140 cccatgcatt caaactga 1158 <210> 62 <211> 385 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Oct4 Amino Acid Sequence <400> 62 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Ala Gly His Leu Ala Ser 20 25 30 Asp Phe Ala Phe Ser Pro Pro Pro Gly Gly Gly Gly Asp Gly Pro Gly 35 40 45 Gly Pro Glu Pro Gly Trp Val Asp Pro Arg Thr Trp Leu Ser Phe Gln 50 55 60 Gly Pro Pro Gly Gly Pro Gly Ile Gly Pro Gly Val Gly Pro Gly Ser 65 70 75 80 Glu Val Trp Gly Ile Pro Pro Cys Pro Pro Pro Tyr Glu Phe Cys Gly 85 90 95 Gly Met Ala Tyr Cys Gly Pro Gln Val Gly Val Gly Leu Val Pro Gln 100 105 110 Gly Gly Leu Glu Thr Ser Gln Pro Glu Gly Glu Ala Gly Val Gly Val 115 120 125 Glu Ser Asn Ser Asp Gly Ala Ser Pro Glu Pro Cys Thr Val Thr Pro 130 135 140 Gly Ala Val Lys Leu Glu Lys Glu Lys Leu Glu Gln Asn Pro Glu Glu 145 150 155 160 Ser Gln Asp Ile Lys Ala Leu Gln Lys Glu Leu Glu Gln Phe Ala Lys 165 170 175 Leu Leu Lys Gln Lys Arg Ile Thr Leu Gly Tyr Thr Gln Ala Asp Val 180 185 190 Gly Leu Thr Leu Gly Val Leu Phe Gly Lys Val Phe Ser Gln Thr Thr 195 200 205 Ile Cys Arg Phe Glu Ala Leu Gln Leu Ser Phe Lys Asn Met Cys Lys 210 215 220 Leu Arg Pro Leu Leu Gln Lys Trp Val Glu Glu Ala Asp Asn Asn Glu 225 230 235 240 Asn Leu Gln Glu Ile Cys Lys Ala Glu Thr Leu Val Gln Ala Arg Lys 245 250 255 Arg Lys Arg Thr Ser Ile Glu Asn Arg Val Arg Gly Asn Leu Glu Asn 260 265 270 Leu Phe Leu Gln Cys Pro Lys Pro Thr Leu Gln Gln Ile Ser His Ile 275 280 285 Ala Gln Gln Leu Gly Leu Glu Lys Asp Val Val Arg Val Trp Phe Cys 290 295 300 Asn Arg Arg Gln Lys Gly Lys Arg Ser Ser Ser Asp Tyr Ala Gln Arg 305 310 315 320 Glu Asp Phe Glu Ala Ala Gly Ser Pro Phe Ser Gly Gly Pro Val Ser 325 330 335 Phe Pro Leu Ala Pro Gly Pro His Phe Gly Thr Pro Gly Tyr Gly Ser 340 345 350 Pro His Phe Thr Ala Leu Tyr Ser Ser Val Pro Phe Pro Glu Gly Glu 355 360 365 Ala Phe Pro Pro Val Ser Val Thr Thr Leu Gly Ser Pro Met His Ser 370 375 380 Asn 385 <210> 63 <211> 1185 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Oct4 cDNA Sequence <400> 63 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctggcggg acacctggct 120 tcggatttcg ccttctcgcc ccctccaggt ggtggaggtg atgggccagg ggggccggag 180 ccgggctggg ttgatcctcg gacctggcta agcttccaag gccctcctgg agggccagga 240 atcgggccgg gggttgggcc aggctctgag gtgtggggga ttcccccatg ccccccgccg 300 tatgagttct gtggggggat ggcgtactgt gggccccagg ttggagtggg gctagtgccc 360 caaggcggct tggagacctc tcagcctgag ggcgaagcag gagtcggggt ggagagcaac 420 tccgatgggg cctccccgga gccctgcacc gtcacccctg gtgccgtgaa gctggagaag 480 gagaagctgg agcaaaaccc ggaggagtcc caggacatca aagctctgca gaaagaactc 540 gagcaatttg ccaagctcct gaagcagaag aggatcaccc tgggatatac acaggccgat 600 gtggggctca ccctgggggt tctatttggg aaggtattca gccaaacgac catctgccgc 660 tttgaggctc tgcagcttag cttcaagaac atgtgtaagc tgcggccctt gctgcagaag 720 tgggtggagg aagctgacaa caatgaaaat cttcaggaga tatgcaaagc agaaaccctc 780 gtgcaggccc gaaagagaaa gcgaaccagt atcgagaacc gagtgagagg caacctggag 840 aatttgttcc tgcagtgccc gaaacccaca ctgcagcaga tcagccacat cgcccagcag 900 cttgggctcg agaaggatgt ggtccgagtg tggttctgta accggcgcca gaagggcaag 960 cgatcaagca gcgactatgc acaacgagag gattttgagg ctgctgggtc tcctttctca 1020 gggggaccag tgtcctttcc tctggcccca gggccccatt ttggtacccc aggctatggg 1080 agccctcact tcactgcact gtactcctcg gtccctttcc ctgaggggga agcctttccc 1140 cctgtctccg tcaccactct gggctctccc atgcattcaa actga 1185 <210> 64 <211> 394 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Oct4 Amino Acid Sequence <400> 64 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Ala Gly His Leu Ala Ser Asp Phe Ala Phe Ser Pro Pro 35 40 45 Pro Gly Gly Gly Gly Asp Gly Pro Gly Gly Pro Glu Pro Gly Trp Val 50 55 60 Asp Pro Arg Thr Trp Leu Ser Phe Gln Gly Pro Pro Gly Gly Pro Gly 65 70 75 80 Ile Gly Pro Gly Val Gly Pro Gly Ser Glu Val Trp Gly Ile Pro Pro 85 90 95 Cys Pro Pro Pro Tyr Glu Phe Cys Gly Gly Met Ala Tyr Cys Gly Pro 100 105 110 Gln Val Gly Val Gly Leu Val Pro Gln Gly Gly Leu Glu Thr Ser Gln 115 120 125 Pro Glu Gly Glu Ala Gly Val Gly Val Glu Ser Asn Ser Asp Gly Ala 130 135 140 Ser Pro Glu Pro Cys Thr Val Thr Pro Gly Ala Val Lys Leu Glu Lys 145 150 155 160 Glu Lys Leu Glu Gln Asn Pro Glu Glu Ser Gln Asp Ile Lys Ala Leu 165 170 175 Gln Lys Glu Leu Glu Gln Phe Ala Lys Leu Leu Lys Gln Lys Arg Ile 180 185 190 Thr Leu Gly Tyr Thr Gln Ala Asp Val Gly Leu Thr Leu Gly Val Leu 195 200 205 Phe Gly Lys Val Phe Ser Gln Thr Thr Ile Cys Arg Phe Glu Ala Leu 210 215 220 Gln Leu Ser Phe Lys Asn Met Cys Lys Leu Arg Pro Leu Leu Gln Lys 225 230 235 240 Trp Val Glu Glu Ala Asp Asn Asn Glu Asn Leu Gln Glu Ile Cys Lys 245 250 255 Ala Glu Thr Leu Val Gln Ala Arg Lys Arg Lys Arg Thr Ser Ile Glu 260 265 270 Asn Arg Val Arg Gly Asn Leu Glu Asn Leu Phe Leu Gln Cys Pro Lys 275 280 285 Pro Thr Leu Gln Gln Ile Ser His Ile Ala Gln Gln Leu Gly Leu Glu 290 295 300 Lys Asp Val Val Arg Val Trp Phe Cys Asn Arg Arg Gln Lys Gly Lys 305 310 315 320 Arg Ser Ser Ser Asp Tyr Ala Gln Arg Glu Asp Phe Glu Ala Ala Gly 325 330 335 Ser Pro Phe Ser Gly Gly Pro Val Ser Phe Pro Leu Ala Pro Gly Pro 340 345 350 His Phe Gly Thr Pro Gly Tyr Gly Ser Pro His Phe Thr Ala Leu Tyr 355 360 365 Ser Ser Val Pro Phe Pro Glu Gly Glu Ala Phe Pro Pro Val Ser Val 370 375 380 Thr Thr Leu Gly Ser Pro Met His Ser Asn 385 390 <210> 65 <211> 1185 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Oct4-JO-84 MTD cDNA Sequence <400> 65 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaaggc gggacacctg gcttcggatt tcgccttctc gccccctcca 120 ggtggtggag gtgatgggcc aggggggccg gagccgggct gggttgatcc tcggacctgg 180 ctaagcttcc aaggccctcc tggagggcca ggaatcgggc cgggggttgg gccaggctct 240 gaggtgtggg ggattccccc atgccccccg ccgtatgagt tctgtggggg gatggcgtac 300 tgtgggcccc aggttggagt ggggctagtg ccccaaggcg gcttggagac ctctcagcct 360 gagggcgaag caggagtcgg ggtggagagc aactccgatg gggcctcccc ggagccctgc 420 accgtcaccc ctggtgccgt gaagctggag aaggagaagc tggagcaaaa cccggaggag 480 tcccaggaca tcaaagctct gcagaaagaa ctcgagcaat ttgccaagct cctgaagcag 540 aagaggatca ccctgggata tacacaggcc gatgtggggc tcaccctggg ggttctattt 600 gggaaggtat tcagccaaac gaccatctgc cgctttgagg ctctgcagct tagcttcaag 660 aacatgtgta agctgcggcc cttgctgcag aagtgggtgg aggaagctga caacaatgaa 720 aatcttcagg agatatgcaa agcagaaacc ctcgtgcagg cccgaaagag aaagcgaacc 780 agtatcgaga accgagtgag aggcaacctg gagaatttgt tcctgcagtg cccgaaaccc 840 acactgcagc agatcagcca catcgcccag cagcttgggc tcgagaagga tgtggtccga 900 gtgtggttct gtaaccggcg ccagaagggc aagcgatcaa gcagcgacta tgcacaacga 960 gaggattttg aggctgctgg gtctcctttc tcagggggac cagtgtcctt tcctctggcc 1020 ccagggcccc attttggtac cccaggctat gggagccctc acttcactgc actgtactcc 1080 tcggtccctt tccctgaggg ggaagccttt ccccctgtct ccgtcaccac tctgggctct 1140 cccatgcatt caaacctggt ggcggcgctg ctggcggtgc tgtga 1185 <210> 66 <211> 394 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Oct4-JO-84 MTD Amino Acid Sequence <400> 66 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Ala Gly His Leu Ala Ser 20 25 30 Asp Phe Ala Phe Ser Pro Pro Pro Gly Gly Gly Gly Asp Gly Pro Gly 35 40 45 Gly Pro Glu Pro Gly Trp Val Asp Pro Arg Thr Trp Leu Ser Phe Gln 50 55 60 Gly Pro Pro Gly Gly Pro Gly Ile Gly Pro Gly Val Gly Pro Gly Ser 65 70 75 80 Glu Val Trp Gly Ile Pro Pro Cys Pro Pro Pro Tyr Glu Phe Cys Gly 85 90 95 Gly Met Ala Tyr Cys Gly Pro Gln Val Gly Val Gly Leu Val Pro Gln 100 105 110 Gly Gly Leu Glu Thr Ser Gln Pro Glu Gly Glu Ala Gly Val Gly Val 115 120 125 Glu Ser Asn Ser Asp Gly Ala Ser Pro Glu Pro Cys Thr Val Thr Pro 130 135 140 Gly Ala Val Lys Leu Glu Lys Glu Lys Leu Glu Gln Asn Pro Glu Glu 145 150 155 160 Ser Gln Asp Ile Lys Ala Leu Gln Lys Glu Leu Glu Gln Phe Ala Lys 165 170 175 Leu Leu Lys Gln Lys Arg Ile Thr Leu Gly Tyr Thr Gln Ala Asp Val 180 185 190 Gly Leu Thr Leu Gly Val Leu Phe Gly Lys Val Phe Ser Gln Thr Thr 195 200 205 Ile Cys Arg Phe Glu Ala Leu Gln Leu Ser Phe Lys Asn Met Cys Lys 210 215 220 Leu Arg Pro Leu Leu Gln Lys Trp Val Glu Glu Ala Asp Asn Asn Glu 225 230 235 240 Asn Leu Gln Glu Ile Cys Lys Ala Glu Thr Leu Val Gln Ala Arg Lys 245 250 255 Arg Lys Arg Thr Ser Ile Glu Asn Arg Val Arg Gly Asn Leu Glu Asn 260 265 270 Leu Phe Leu Gln Cys Pro Lys Pro Thr Leu Gln Gln Ile Ser His Ile 275 280 285 Ala Gln Gln Leu Gly Leu Glu Lys Asp Val Val Arg Val Trp Phe Cys 290 295 300 Asn Arg Arg Gln Lys Gly Lys Arg Ser Ser Ser Asp Tyr Ala Gln Arg 305 310 315 320 Glu Asp Phe Glu Ala Ala Gly Ser Pro Phe Ser Gly Gly Pro Val Ser 325 330 335 Phe Pro Leu Ala Pro Gly Pro His Phe Gly Thr Pro Gly Tyr Gly Ser 340 345 350 Pro His Phe Thr Ala Leu Tyr Ser Ser Val Pro Phe Pro Glu Gly Glu 355 360 365 Ala Phe Pro Pro Val Ser Val Thr Thr Leu Gly Ser Pro Met His Ser 370 375 380 Asn Leu Val Ala Ala Leu Leu Ala Val Leu 385 390 <210> 67 <211> 1212 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Oct4-JO-84 MTD cDNA Sequence <400> 67 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctggcggg acacctggct 120 tcggatttcg ccttctcgcc ccctccaggt ggtggaggtg atgggccagg ggggccggag 180 ccgggctggg ttgatcctcg gacctggcta agcttccaag gccctcctgg agggccagga 240 atcgggccgg gggttgggcc aggctctgag gtgtggggga ttcccccatg ccccccgccg 300 tatgagttct gtggggggat ggcgtactgt gggccccagg ttggagtggg gctagtgccc 360 caaggcggct tggagacctc tcagcctgag ggcgaagcag gagtcggggt ggagagcaac 420 tccgatgggg cctccccgga gccctgcacc gtcacccctg gtgccgtgaa gctggagaag 480 gagaagctgg agcaaaaccc ggaggagtcc caggacatca aagctctgca gaaagaactc 540 gagcaatttg ccaagctcct gaagcagaag aggatcaccc tgggatatac acaggccgat 600 gtggggctca ccctgggggt tctatttggg aaggtattca gccaaacgac catctgccgc 660 tttgaggctc tgcagcttag cttcaagaac atgtgtaagc tgcggccctt gctgcagaag 720 tgggtggagg aagctgacaa caatgaaaat cttcaggaga tatgcaaagc agaaaccctc 780 gtgcaggccc gaaagagaaa gcgaaccagt atcgagaacc gagtgagagg caacctggag 840 aatttgttcc tgcagtgccc gaaacccaca ctgcagcaga tcagccacat cgcccagcag 900 cttgggctcg agaaggatgt ggtccgagtg tggttctgta accggcgcca gaagggcaag 960 cgatcaagca gcgactatgc acaacgagag gattttgagg ctgctgggtc tcctttctca 1020 gggggaccag tgtcctttcc tctggcccca gggccccatt ttggtacccc aggctatggg 1080 agccctcact tcactgcact gtactcctcg gtccctttcc ctgaggggga agcctttccc 1140 cctgtctccg tcaccactct gggctctccc atgcattcaa acctggtggc ggcgctgctg 1200 gcggtgctgt ga 1212 <210> 68 <211> 403 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Oct4-JO-84 MTD Amino Acid Sequence <400> 68 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Ala Gly His Leu Ala Ser Asp Phe Ala Phe Ser Pro Pro 35 40 45 Pro Gly Gly Gly Gly Asp Gly Pro Gly Gly Pro Glu Pro Gly Trp Val 50 55 60 Asp Pro Arg Thr Trp Leu Ser Phe Gln Gly Pro Pro Gly Gly Pro Gly 65 70 75 80 Ile Gly Pro Gly Val Gly Pro Gly Ser Glu Val Trp Gly Ile Pro Pro 85 90 95 Cys Pro Pro Pro Tyr Glu Phe Cys Gly Gly Met Ala Tyr Cys Gly Pro 100 105 110 Gln Val Gly Val Gly Leu Val Pro Gln Gly Gly Leu Glu Thr Ser Gln 115 120 125 Pro Glu Gly Glu Ala Gly Val Gly Val Glu Ser Asn Ser Asp Gly Ala 130 135 140 Ser Pro Glu Pro Cys Thr Val Thr Pro Gly Ala Val Lys Leu Glu Lys 145 150 155 160 Glu Lys Leu Glu Gln Asn Pro Glu Glu Ser Gln Asp Ile Lys Ala Leu 165 170 175 Gln Lys Glu Leu Glu Gln Phe Ala Lys Leu Leu Lys Gln Lys Arg Ile 180 185 190 Thr Leu Gly Tyr Thr Gln Ala Asp Val Gly Leu Thr Leu Gly Val Leu 195 200 205 Phe Gly Lys Val Phe Ser Gln Thr Thr Ile Cys Arg Phe Glu Ala Leu 210 215 220 Gln Leu Ser Phe Lys Asn Met Cys Lys Leu Arg Pro Leu Leu Gln Lys 225 230 235 240 Trp Val Glu Glu Ala Asp Asn Asn Glu Asn Leu Gln Glu Ile Cys Lys 245 250 255 Ala Glu Thr Leu Val Gln Ala Arg Lys Arg Lys Arg Thr Ser Ile Glu 260 265 270 Asn Arg Val Arg Gly Asn Leu Glu Asn Leu Phe Leu Gln Cys Pro Lys 275 280 285 Pro Thr Leu Gln Gln Ile Ser His Ile Ala Gln Gln Leu Gly Leu Glu 290 295 300 Lys Asp Val Val Arg Val Trp Phe Cys Asn Arg Arg Gln Lys Gly Lys 305 310 315 320 Arg Ser Ser Ser Asp Tyr Ala Gln Arg Glu Asp Phe Glu Ala Ala Gly 325 330 335 Ser Pro Phe Ser Gly Gly Pro Val Ser Phe Pro Leu Ala Pro Gly Pro 340 345 350 His Phe Gly Thr Pro Gly Tyr Gly Ser Pro His Phe Thr Ala Leu Tyr 355 360 365 Ser Ser Val Pro Phe Pro Glu Gly Glu Ala Phe Pro Pro Val Ser Val 370 375 380 Thr Thr Leu Gly Ser Pro Met His Ser Asn Leu Val Ala Ala Leu Leu 385 390 395 400 Ala Val Leu <210> 69 <211> 1182 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Oct4 cDNA Sequence <400> 69 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cggcgggaca cctggcttcg 120 gatttcgcct tctcgccccc tccaggtggt ggaggtgatg ggccaggggg gccggagccg 180 ggctgggttg atcctcggac ctggctaagc ttccaaggcc ctcctggagg gccaggaatc 240 gggccggggg ttgggccagg ctctgaggtg tgggggattc ccccatgccc cccgccgtat 300 gagttctgtg gggggatggc gtactgtggg ccccaggttg gagtggggct agtgccccaa 360 ggcggcttgg agacctctca gcctgagggc gaagcaggag tcggggtgga gagcaactcc 420 gatggggcct ccccggagcc ctgcaccgtc acccctggtg ccgtgaagct ggagaaggag 480 aagctggagc aaaacccgga ggagtcccag gacatcaaag ctctgcagaa agaactcgag 540 caatttgcca agctcctgaa gcagaagagg atcaccctgg gatatacaca ggccgatgtg 600 gggctcaccc tgggggttct atttgggaag gtattcagcc aaacgaccat ctgccgcttt 660 gaggctctgc agcttagctt caagaacatg tgtaagctgc ggcccttgct gcagaagtgg 720 gtggaggaag ctgacaacaa tgaaaatctt caggagatat gcaaagcaga aaccctcgtg 780 caggcccgaa agagaaagcg aaccagtatc gagaaccgag tgagaggcaa cctggagaat 840 ttgttcctgc agtgcccgaa acccacactg cagcagatca gccacatcgc ccagcagctt 900 gggctcgaga aggatgtggt ccgagtgtgg ttctgtaacc ggcgccagaa gggcaagcga 960 tcaagcagcg actatgcaca acgagaggat tttgaggctg ctgggtctcc tttctcaggg 1020 ggaccagtgt cctttcctct ggccccaggg ccccattttg gtaccccagg ctatgggagc 1080 cctcacttca ctgcactgta ctcctcggtc cctttccctg agggggaagc ctttccccct 1140 gtctccgtca ccactctggg ctctcccatg cattcaaact ga 1182 <210> 70 <211> 393 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Oct4 Amico Acid Sequence <400> 70 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Ala Gly His Leu Ala Ser Asp Phe Ala Phe Ser Pro Pro Pro 35 40 45 Gly Gly Gly Gly Asp Gly Pro Gly Gly Pro Glu Pro Gly Trp Val Asp 50 55 60 Pro Arg Thr Trp Leu Ser Phe Gln Gly Pro Pro Gly Gly Pro Gly Ile 65 70 75 80 Gly Pro Gly Val Gly Pro Gly Ser Glu Val Trp Gly Ile Pro Pro Cys 85 90 95 Pro Pro Pro Tyr Glu Phe Cys Gly Gly Met Ala Tyr Cys Gly Pro Gln 100 105 110 Val Gly Val Gly Leu Val Pro Gln Gly Gly Leu Glu Thr Ser Gln Pro 115 120 125 Glu Gly Glu Ala Gly Val Gly Val Glu Ser Asn Ser Asp Gly Ala Ser 130 135 140 Pro Glu Pro Cys Thr Val Thr Pro Gly Ala Val Lys Leu Glu Lys Glu 145 150 155 160 Lys Leu Glu Gln Asn Pro Glu Glu Ser Gln Asp Ile Lys Ala Leu Gln 165 170 175 Lys Glu Leu Glu Gln Phe Ala Lys Leu Leu Lys Gln Lys Arg Ile Thr 180 185 190 Leu Gly Tyr Thr Gln Ala Asp Val Gly Leu Thr Leu Gly Val Leu Phe 195 200 205 Gly Lys Val Phe Ser Gln Thr Thr Ile Cys Arg Phe Glu Ala Leu Gln 210 215 220 Leu Ser Phe Lys Asn Met Cys Lys Leu Arg Pro Leu Leu Gln Lys Trp 225 230 235 240 Val Glu Glu Ala Asp Asn Asn Glu Asn Leu Gln Glu Ile Cys Lys Ala 245 250 255 Glu Thr Leu Val Gln Ala Arg Lys Arg Lys Arg Thr Ser Ile Glu Asn 260 265 270 Arg Val Arg Gly Asn Leu Glu Asn Leu Phe Leu Gln Cys Pro Lys Pro 275 280 285 Thr Leu Gln Gln Ile Ser His Ile Ala Gln Gln Leu Gly Leu Glu Lys 290 295 300 Asp Val Val Arg Val Trp Phe Cys Asn Arg Arg Gln Lys Gly Lys Arg 305 310 315 320 Ser Ser Ser Asp Tyr Ala Gln Arg Glu Asp Phe Glu Ala Ala Gly Ser 325 330 335 Pro Phe Ser Gly Gly Pro Val Ser Phe Pro Leu Ala Pro Gly Pro His 340 345 350 Phe Gly Thr Pro Gly Tyr Gly Ser Pro His Phe Thr Ala Leu Tyr Ser 355 360 365 Ser Val Pro Phe Pro Glu Gly Glu Ala Phe Pro Pro Val Ser Val Thr 370 375 380 Thr Leu Gly Ser Pro Met His Ser Asn 385 390 <210> 71 <211> 1182 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Oct4-JO-86 MTD cDNA Sequence <400> 71 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaaggc gggacacctg gcttcggatt tcgccttctc gccccctcca 120 ggtggtggag gtgatgggcc aggggggccg gagccgggct gggttgatcc tcggacctgg 180 ctaagcttcc aaggccctcc tggagggcca ggaatcgggc cgggggttgg gccaggctct 240 gaggtgtggg ggattccccc atgccccccg ccgtatgagt tctgtggggg gatggcgtac 300 tgtgggcccc aggttggagt ggggctagtg ccccaaggcg gcttggagac ctctcagcct 360 gagggcgaag caggagtcgg ggtggagagc aactccgatg gggcctcccc ggagccctgc 420 accgtcaccc ctggtgccgt gaagctggag aaggagaagc tggagcaaaa cccggaggag 480 tcccaggaca tcaaagctct gcagaaagaa ctcgagcaat ttgccaagct cctgaagcag 540 aagaggatca ccctgggata tacacaggcc gatgtggggc tcaccctggg ggttctattt 600 gggaaggtat tcagccaaac gaccatctgc cgctttgagg ctctgcagct tagcttcaag 660 aacatgtgta agctgcggcc cttgctgcag aagtgggtgg aggaagctga caacaatgaa 720 aatcttcagg agatatgcaa agcagaaacc ctcgtgcagg cccgaaagag aaagcgaacc 780 agtatcgaga accgagtgag aggcaacctg gagaatttgt tcctgcagtg cccgaaaccc 840 acactgcagc agatcagcca catcgcccag cagcttgggc tcgagaagga tgtggtccga 900 gtgtggttct gtaaccggcg ccagaagggc aagcgatcaa gcagcgacta tgcacaacga 960 gaggattttg aggctgctgg gtctcctttc tcagggggac cagtgtcctt tcctctggcc 1020 ccagggcccc attttggtac cccaggctat gggagccctc acttcactgc actgtactcc 1080 tcggtccctt tccctgaggg ggaagccttt ccccctgtct ccgtcaccac tctgggctct 1140 cccatgcatt caaacctggc ggtgctggcg gcggcgccgt ga 1182 <210> 72 <211> 393 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Oct4-JO-86 MTD Amino Acid Sequence <400> 72 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Ala Gly His Leu Ala Ser 20 25 30 Asp Phe Ala Phe Ser Pro Pro Pro Gly Gly Gly Gly Asp Gly Pro Gly 35 40 45 Gly Pro Glu Pro Gly Trp Val Asp Pro Arg Thr Trp Leu Ser Phe Gln 50 55 60 Gly Pro Pro Gly Gly Pro Gly Ile Gly Pro Gly Val Gly Pro Gly Ser 65 70 75 80 Glu Val Trp Gly Ile Pro Pro Cys Pro Pro Pro Tyr Glu Phe Cys Gly 85 90 95 Gly Met Ala Tyr Cys Gly Pro Gln Val Gly Val Gly Leu Val Pro Gln 100 105 110 Gly Gly Leu Glu Thr Ser Gln Pro Glu Gly Glu Ala Gly Val Gly Val 115 120 125 Glu Ser Asn Ser Asp Gly Ala Ser Pro Glu Pro Cys Thr Val Thr Pro 130 135 140 Gly Ala Val Lys Leu Glu Lys Glu Lys Leu Glu Gln Asn Pro Glu Glu 145 150 155 160 Ser Gln Asp Ile Lys Ala Leu Gln Lys Glu Leu Glu Gln Phe Ala Lys 165 170 175 Leu Leu Lys Gln Lys Arg Ile Thr Leu Gly Tyr Thr Gln Ala Asp Val 180 185 190 Gly Leu Thr Leu Gly Val Leu Phe Gly Lys Val Phe Ser Gln Thr Thr 195 200 205 Ile Cys Arg Phe Glu Ala Leu Gln Leu Ser Phe Lys Asn Met Cys Lys 210 215 220 Leu Arg Pro Leu Leu Gln Lys Trp Val Glu Glu Ala Asp Asn Asn Glu 225 230 235 240 Asn Leu Gln Glu Ile Cys Lys Ala Glu Thr Leu Val Gln Ala Arg Lys 245 250 255 Arg Lys Arg Thr Ser Ile Glu Asn Arg Val Arg Gly Asn Leu Glu Asn 260 265 270 Leu Phe Leu Gln Cys Pro Lys Pro Thr Leu Gln Gln Ile Ser His Ile 275 280 285 Ala Gln Gln Leu Gly Leu Glu Lys Asp Val Val Arg Val Trp Phe Cys 290 295 300 Asn Arg Arg Gln Lys Gly Lys Arg Ser Ser Ser Asp Tyr Ala Gln Arg 305 310 315 320 Glu Asp Phe Glu Ala Ala Gly Ser Pro Phe Ser Gly Gly Pro Val Ser 325 330 335 Phe Pro Leu Ala Pro Gly Pro His Phe Gly Thr Pro Gly Tyr Gly Ser 340 345 350 Pro His Phe Thr Ala Leu Tyr Ser Ser Val Pro Phe Pro Glu Gly Glu 355 360 365 Ala Phe Pro Pro Val Ser Val Thr Thr Leu Gly Ser Pro Met His Ser 370 375 380 Asn Leu Ala Val Leu Ala Ala Ala Pro 385 390 <210> 73 <211> 1206 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Oct4-JO-86 MTD cDNA Sequence <400> 73 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cggcgggaca cctggcttcg 120 gatttcgcct tctcgccccc tccaggtggt ggaggtgatg ggccaggggg gccggagccg 180 ggctgggttg atcctcggac ctggctaagc ttccaaggcc ctcctggagg gccaggaatc 240 gggccggggg ttgggccagg ctctgaggtg tgggggattc ccccatgccc cccgccgtat 300 gagttctgtg gggggatggc gtactgtggg ccccaggttg gagtggggct agtgccccaa 360 ggcggcttgg agacctctca gcctgagggc gaagcaggag tcggggtgga gagcaactcc 420 gatggggcct ccccggagcc ctgcaccgtc acccctggtg ccgtgaagct ggagaaggag 480 aagctggagc aaaacccgga ggagtcccag gacatcaaag ctctgcagaa agaactcgag 540 caatttgcca agctcctgaa gcagaagagg atcaccctgg gatatacaca ggccgatgtg 600 gggctcaccc tgggggttct atttgggaag gtattcagcc aaacgaccat ctgccgcttt 660 gaggctctgc agcttagctt caagaacatg tgtaagctgc ggcccttgct gcagaagtgg 720 gtggaggaag ctgacaacaa tgaaaatctt caggagatat gcaaagcaga aaccctcgtg 780 caggcccgaa agagaaagcg aaccagtatc gagaaccgag tgagaggcaa cctggagaat 840 ttgttcctgc agtgcccgaa acccacactg cagcagatca gccacatcgc ccagcagctt 900 gggctcgaga aggatgtggt ccgagtgtgg ttctgtaacc ggcgccagaa gggcaagcga 960 tcaagcagcg actatgcaca acgagaggat tttgaggctg ctgggtctcc tttctcaggg 1020 ggaccagtgt cctttcctct ggccccaggg ccccattttg gtaccccagg ctatgggagc 1080 cctcacttca ctgcactgta ctcctcggtc cctttccctg agggggaagc ctttccccct 1140 gtctccgtca ccactctggg ctctcccatg cattcaaacc tggcggtgct ggcggcggcg 1200 ccgtga 1206 <210> 74 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Oct4-JO-86 MTD Amino Acid Sequence <400> 74 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Ala Gly His Leu Ala Ser Asp Phe Ala Phe Ser Pro Pro Pro 35 40 45 Gly Gly Gly Gly Asp Gly Pro Gly Gly Pro Glu Pro Gly Trp Val Asp 50 55 60 Pro Arg Thr Trp Leu Ser Phe Gln Gly Pro Pro Gly Gly Pro Gly Ile 65 70 75 80 Gly Pro Gly Val Gly Pro Gly Ser Glu Val Trp Gly Ile Pro Pro Cys 85 90 95 Pro Pro Pro Tyr Glu Phe Cys Gly Gly Met Ala Tyr Cys Gly Pro Gln 100 105 110 Val Gly Val Gly Leu Val Pro Gln Gly Gly Leu Glu Thr Ser Gln Pro 115 120 125 Glu Gly Glu Ala Gly Val Gly Val Glu Ser Asn Ser Asp Gly Ala Ser 130 135 140 Pro Glu Pro Cys Thr Val Thr Pro Gly Ala Val Lys Leu Glu Lys Glu 145 150 155 160 Lys Leu Glu Gln Asn Pro Glu Glu Ser Gln Asp Ile Lys Ala Leu Gln 165 170 175 Lys Glu Leu Glu Gln Phe Ala Lys Leu Leu Lys Gln Lys Arg Ile Thr 180 185 190 Leu Gly Tyr Thr Gln Ala Asp Val Gly Leu Thr Leu Gly Val Leu Phe 195 200 205 Gly Lys Val Phe Ser Gln Thr Thr Ile Cys Arg Phe Glu Ala Leu Gln 210 215 220 Leu Ser Phe Lys Asn Met Cys Lys Leu Arg Pro Leu Leu Gln Lys Trp 225 230 235 240 Val Glu Glu Ala Asp Asn Asn Glu Asn Leu Gln Glu Ile Cys Lys Ala 245 250 255 Glu Thr Leu Val Gln Ala Arg Lys Arg Lys Arg Thr Ser Ile Glu Asn 260 265 270 Arg Val Arg Gly Asn Leu Glu Asn Leu Phe Leu Gln Cys Pro Lys Pro 275 280 285 Thr Leu Gln Gln Ile Ser His Ile Ala Gln Gln Leu Gly Leu Glu Lys 290 295 300 Asp Val Val Arg Val Trp Phe Cys Asn Arg Arg Gln Lys Gly Lys Arg 305 310 315 320 Ser Ser Ser Asp Tyr Ala Gln Arg Glu Asp Phe Glu Ala Ala Gly Ser 325 330 335 Pro Phe Ser Gly Gly Pro Val Ser Phe Pro Leu Ala Pro Gly Pro His 340 345 350 Phe Gly Thr Pro Gly Tyr Gly Ser Pro His Phe Thr Ala Leu Tyr Ser 355 360 365 Ser Val Pro Phe Pro Glu Gly Glu Ala Phe Pro Pro Val Ser Val Thr 370 375 380 Thr Leu Gly Ser Pro Met His Ser Asn Leu Ala Val Leu Ala Ala Ala 385 390 395 400 Pro <210> 75 <211> 969 <212> DNA <213> Artificial Sequence <220> <223> NLS-Sox2 cDNA Sequence <400> 75 atgaagaaga agaggaagta caacatgatg gagacggagc tgaagccgcc gggcccgcag 60 caaacttcgg ggggcggcgg cggcaactcc accgcggcgg cggccggcgg caaccagaaa 120 aacagcccgg accgcgtcaa gcggcccatg aatgccttca tggtgtggtc ccgcgggcag 180 cggcgcaaga tggcccagga gaaccccaag atgcacaact cggagatcag caagcgcctg 240 ggcgccgagt ggaaactttt gtcggagacg gagaagcggc cgttcatcga cgaggctaag 300 cggctgcgag cgctgcacat gaaggagcac ccggattata aataccggcc ccggcggaaa 360 accaagacgc tcatgaagaa ggataagtac acgctgcccg gcgggctgct ggcccccggc 420 ggcaatagca tggcgagcgg ggtcggggtg ggcgccggcc tgggcgcggg cgtgaaccag 480 cgcatggaca gttacgcgca catgaacggc tggagcaacg gcagctacag catgatgcag 540 gaccagctgg gctacccgca gcacccgggc ctcaatgcgc acggcgcagc gcagatgcag 600 cccatgcacc gctacgacgt gagcgccctg cagtacaact ccatgaccag ctcgcagacc 660 tacatgaacg gctcgcccac ctacagcatg tcctactcgc agcagggcac ccctggcatg 720 gctcttggct ccatgggttc ggtggtcaag tccgaggcca gctccagccc ccctgtggtt 780 acctcttcct cccactccag ggcgccctgc caggccgggg acctccggga catgatcagc 840 atgtatctcc ccggcgccga ggtgccggaa cccgccgccc ccagcagact tcacatgtcc 900 cagcactacc agagcggccc ggtgcccggc acggccatta acggcacact gcccctctca 960 cacatgtga 969 <210> 76 <211> 322 <212> PRT <213> Artificial Sequence <220> <223> NLS-Sox2 Amino Acid Sequence <400> 76 Met Lys Lys Lys Arg Lys Tyr Asn Met Met Glu Thr Glu Leu Lys Pro 1 5 10 15 Pro Gly Pro Gln Gln Thr Ser Gly Gly Gly Gly Gly Asn Ser Thr Ala 20 25 30 Ala Ala Ala Gly Gly Asn Gln Lys Asn Ser Pro Asp Arg Val Lys Arg 35 40 45 Pro Met Asn Ala Phe Met Val Trp Ser Arg Gly Gln Arg Arg Lys Met 50 55 60 Ala Gln Glu Asn Pro Lys Met His Asn Ser Glu Ile Ser Lys Arg Leu 65 70 75 80 Gly Ala Glu Trp Lys Leu Leu Ser Glu Thr Glu Lys Arg Pro Phe Ile 85 90 95 Asp Glu Ala Lys Arg Leu Arg Ala Leu His Met Lys Glu His Pro Asp 100 105 110 Tyr Lys Tyr Arg Pro Arg Arg Lys Thr Lys Thr Leu Met Lys Lys Asp 115 120 125 Lys Tyr Thr Leu Pro Gly Gly Leu Leu Ala Pro Gly Gly Asn Ser Met 130 135 140 Ala Ser Gly Val Gly Val Gly Ala Gly Leu Gly Ala Gly Val Asn Gln 145 150 155 160 Arg Met Asp Ser Tyr Ala His Met Asn Gly Trp Ser Asn Gly Ser Tyr 165 170 175 Ser Met Met Gln Asp Gln Leu Gly Tyr Pro Gln His Pro Gly Leu Asn 180 185 190 Ala His Gly Ala Ala Gln Met Gln Pro Met His Arg Tyr Asp Val Ser 195 200 205 Ala Leu Gln Tyr Asn Ser Met Thr Ser Ser Gln Thr Tyr Met Asn Gly 210 215 220 Ser Pro Thr Tyr Ser Met Ser Tyr Ser Gln Gln Gly Thr Pro Gly Met 225 230 235 240 Ala Leu Gly Ser Met Gly Ser Val Val Lys Ser Glu Ala Ser Ser Ser 245 250 255 Pro Pro Val Val Thr Ser Ser Ser His Ser Arg Ala Pro Cys Gln Ala 260 265 270 Gly Asp Leu Arg Asp Met Ile Ser Met Tyr Leu Pro Gly Ala Glu Val 275 280 285 Pro Glu Pro Ala Ala Pro Ser Arg Leu His Met Ser Gln His Tyr Gln 290 295 300 Ser Gly Pro Val Pro Gly Thr Ala Ile Asn Gly Thr Leu Pro Leu Ser 305 310 315 320 His Met <210> 77 <211> 996 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Sox2 cDNA Sequence <400> 77 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgtacaa catgatggag 60 acggagctga agccgccggg cccgcagcaa acttcggggg gcggcggcgg caactccacc 120 gcggcggcgg ccggcggcaa ccagaaaaac agcccggacc gcgtcaagcg gcccatgaat 180 gccttcatgg tgtggtcccg cgggcagcgg cgcaagatgg cccaggagaa ccccaagatg 240 cacaactcgg agatcagcaa gcgcctgggc gccgagtgga aacttttgtc ggagacggag 300 aagcggccgt tcatcgacga ggctaagcgg ctgcgagcgc tgcacatgaa ggagcacccg 360 gattataaat accggccccg gcggaaaacc aagacgctca tgaagaagga taagtacacg 420 ctgcccggcg ggctgctggc ccccggcggc aatagcatgg cgagcggggt cggggtgggc 480 gccggcctgg gcgcgggcgt gaaccagcgc atggacagtt acgcgcacat gaacggctgg 540 agcaacggca gctacagcat gatgcaggac cagctgggct acccgcagca cccgggcctc 600 aatgcgcacg gcgcagcgca gatgcagccc atgcaccgct acgacgtgag cgccctgcag 660 tacaactcca tgaccagctc gcagacctac atgaacggct cgcccaccta cagcatgtcc 720 tactcgcagc agggcacccc tggcatggct cttggctcca tgggttcggt ggtcaagtcc 780 gaggccagct ccagcccccc tgtggttacc tcttcctccc actccagggc gccctgccag 840 gccggggacc tccgggacat gatcagcatg tatctccccg gcgccgaggt gccggaaccc 900 gccgccccca gcagacttca catgtcccag cactaccaga gcggcccggt gcccggcacg 960 gccattaacg gcacactgcc cctctcacac atgtga 996 <210> 78 <211> 331 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Sox2 Amino Acid Sequence <400> 78 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Tyr 1 5 10 15 Asn Met Met Glu Thr Glu Leu Lys Pro Pro Gly Pro Gln Gln Thr Ser 20 25 30 Gly Gly Gly Gly Gly Asn Ser Thr Ala Ala Ala Ala Gly Gly Asn Gln 35 40 45 Lys Asn Ser Pro Asp Arg Val Lys Arg Pro Met Asn Ala Phe Met Val 50 55 60 Trp Ser Arg Gly Gln Arg Arg Lys Met Ala Gln Glu Asn Pro Lys Met 65 70 75 80 His Asn Ser Glu Ile Ser Lys Arg Leu Gly Ala Glu Trp Lys Leu Leu 85 90 95 Ser Glu Thr Glu Lys Arg Pro Phe Ile Asp Glu Ala Lys Arg Leu Arg 100 105 110 Ala Leu His Met Lys Glu His Pro Asp Tyr Lys Tyr Arg Pro Arg Arg 115 120 125 Lys Thr Lys Thr Leu Met Lys Lys Asp Lys Tyr Thr Leu Pro Gly Gly 130 135 140 Leu Leu Ala Pro Gly Gly Asn Ser Met Ala Ser Gly Val Gly Val Gly 145 150 155 160 Ala Gly Leu Gly Ala Gly Val Asn Gln Arg Met Asp Ser Tyr Ala His 165 170 175 Met Asn Gly Trp Ser Asn Gly Ser Tyr Ser Met Met Gln Asp Gln Leu 180 185 190 Gly Tyr Pro Gln His Pro Gly Leu Asn Ala His Gly Ala Ala Gln Met 195 200 205 Gln Pro Met His Arg Tyr Asp Val Ser Ala Leu Gln Tyr Asn Ser Met 210 215 220 Thr Ser Ser Gln Thr Tyr Met Asn Gly Ser Pro Thr Tyr Ser Met Ser 225 230 235 240 Tyr Ser Gln Gln Gly Thr Pro Gly Met Ala Leu Gly Ser Met Gly Ser 245 250 255 Val Val Lys Ser Glu Ala Ser Ser Ser Pro Pro Val Val Thr Ser Ser 260 265 270 Ser His Ser Arg Ala Pro Cys Gln Ala Gly Asp Leu Arg Asp Met Ile 275 280 285 Ser Met Tyr Leu Pro Gly Ala Glu Val Pro Glu Pro Ala Ala Pro Ser 290 295 300 Arg Leu His Met Ser Gln His Tyr Gln Ser Gly Pro Val Pro Gly Thr 305 310 315 320 Ala Ile Asn Gly Thr Leu Pro Leu Ser His Met 325 330 <210> 79 <211> 996 <212> DNA <213> Artificial Sequence <220> <223> NLS-Sox2-JO-84 MTD cDNA Sequence <400> 79 atgaagaaga agaggaagta caacatgatg gagacggagc tgaagccgcc gggcccgcag 60 caaacttcgg ggggcggcgg cggcaactcc accgcggcgg cggccggcgg caaccagaaa 120 aacagcccgg accgcgtcaa gcggcccatg aatgccttca tggtgtggtc ccgcgggcag 180 cggcgcaaga tggcccagga gaaccccaag atgcacaact cggagatcag caagcgcctg 240 ggcgccgagt ggaaactttt gtcggagacg gagaagcggc cgttcatcga cgaggctaag 300 cggctgcgag cgctgcacat gaaggagcac ccggattata aataccggcc ccggcggaaa 360 accaagacgc tcatgaagaa ggataagtac acgctgcccg gcgggctgct ggcccccggc 420 ggcaatagca tggcgagcgg ggtcggggtg ggcgccggcc tgggcgcggg cgtgaaccag 480 cgcatggaca gttacgcgca catgaacggc tggagcaacg gcagctacag catgatgcag 540 gaccagctgg gctacccgca gcacccgggc ctcaatgcgc acggcgcagc gcagatgcag 600 cccatgcacc gctacgacgt gagcgccctg cagtacaact ccatgaccag ctcgcagacc 660 tacatgaacg gctcgcccac ctacagcatg tcctactcgc agcagggcac ccctggcatg 720 gctcttggct ccatgggttc ggtggtcaag tccgaggcca gctccagccc ccctgtggtt 780 acctcttcct cccactccag ggcgccctgc caggccgggg acctccggga catgatcagc 840 atgtatctcc ccggcgccga ggtgccggaa cccgccgccc ccagcagact tcacatgtcc 900 cagcactacc agagcggccc ggtgcccggc acggccatta acggcacact gcccctctca 960 cacatgctgg tggcggcgct gctggcggtg ctgtga 996 <210> 80 <211> 331 <212> PRT <213> Artificial Sequence <220> <223> NLS-Sox2-JO-84 MTD Amino Acid Sequence <400> 80 Met Lys Lys Lys Arg Lys Tyr Asn Met Met Glu Thr Glu Leu Lys Pro 1 5 10 15 Pro Gly Pro Gln Gln Thr Ser Gly Gly Gly Gly Gly Asn Ser Thr Ala 20 25 30 Ala Ala Ala Gly Gly Asn Gln Lys Asn Ser Pro Asp Arg Val Lys Arg 35 40 45 Pro Met Asn Ala Phe Met Val Trp Ser Arg Gly Gln Arg Arg Lys Met 50 55 60 Ala Gln Glu Asn Pro Lys Met His Asn Ser Glu Ile Ser Lys Arg Leu 65 70 75 80 Gly Ala Glu Trp Lys Leu Leu Ser Glu Thr Glu Lys Arg Pro Phe Ile 85 90 95 Asp Glu Ala Lys Arg Leu Arg Ala Leu His Met Lys Glu His Pro Asp 100 105 110 Tyr Lys Tyr Arg Pro Arg Arg Lys Thr Lys Thr Leu Met Lys Lys Asp 115 120 125 Lys Tyr Thr Leu Pro Gly Gly Leu Leu Ala Pro Gly Gly Asn Ser Met 130 135 140 Ala Ser Gly Val Gly Val Gly Ala Gly Leu Gly Ala Gly Val Asn Gln 145 150 155 160 Arg Met Asp Ser Tyr Ala His Met Asn Gly Trp Ser Asn Gly Ser Tyr 165 170 175 Ser Met Met Gln Asp Gln Leu Gly Tyr Pro Gln His Pro Gly Leu Asn 180 185 190 Ala His Gly Ala Ala Gln Met Gln Pro Met His Arg Tyr Asp Val Ser 195 200 205 Ala Leu Gln Tyr Asn Ser Met Thr Ser Ser Gln Thr Tyr Met Asn Gly 210 215 220 Ser Pro Thr Tyr Ser Met Ser Tyr Ser Gln Gln Gly Thr Pro Gly Met 225 230 235 240 Ala Leu Gly Ser Met Gly Ser Val Val Lys Ser Glu Ala Ser Ser Ser 245 250 255 Pro Pro Val Val Thr Ser Ser Ser His Ser Arg Ala Pro Cys Gln Ala 260 265 270 Gly Asp Leu Arg Asp Met Ile Ser Met Tyr Leu Pro Gly Ala Glu Val 275 280 285 Pro Glu Pro Ala Ala Pro Ser Arg Leu His Met Ser Gln His Tyr Gln 290 295 300 Ser Gly Pro Val Pro Gly Thr Ala Ile Asn Gly Thr Leu Pro Leu Ser 305 310 315 320 His Met Leu Val Ala Ala Leu Leu Ala Val Leu 325 330 <210> 81 <211> 1023 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Sox2-JO-84 MTD cDNA Sequence <400> 81 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgtacaa catgatggag 60 acggagctga agccgccggg cccgcagcaa acttcggggg gcggcggcgg caactccacc 120 gcggcggcgg ccggcggcaa ccagaaaaac agcccggacc gcgtcaagcg gcccatgaat 180 gccttcatgg tgtggtcccg cgggcagcgg cgcaagatgg cccaggagaa ccccaagatg 240 cacaactcgg agatcagcaa gcgcctgggc gccgagtgga aacttttgtc ggagacggag 300 aagcggccgt tcatcgacga ggctaagcgg ctgcgagcgc tgcacatgaa ggagcacccg 360 gattataaat accggccccg gcggaaaacc aagacgctca tgaagaagga taagtacacg 420 ctgcccggcg ggctgctggc ccccggcggc aatagcatgg cgagcggggt cggggtgggc 480 gccggcctgg gcgcgggcgt gaaccagcgc atggacagtt acgcgcacat gaacggctgg 540 agcaacggca gctacagcat gatgcaggac cagctgggct acccgcagca cccgggcctc 600 aatgcgcacg gcgcagcgca gatgcagccc atgcaccgct acgacgtgag cgccctgcag 660 tacaactcca tgaccagctc gcagacctac atgaacggct cgcccaccta cagcatgtcc 720 tactcgcagc agggcacccc tggcatggct cttggctcca tgggttcggt ggtcaagtcc 780 gaggccagct ccagcccccc tgtggttacc tcttcctccc actccagggc gccctgccag 840 gccggggacc tccgggacat gatcagcatg tatctccccg gcgccgaggt gccggaaccc 900 gccgccccca gcagacttca catgtcccag cactaccaga gcggcccggt gcccggcacg 960 gccattaacg gcacactgcc cctctcacac atgctggtgg cggcgctgct ggcggtgctg 1020 tga 1023 <210> 82 <211> 340 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Sox2-JO-84 MTD Amino Acid Sequence <400> 82 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Tyr 1 5 10 15 Asn Met Met Glu Thr Glu Leu Lys Pro Pro Gly Pro Gln Gln Thr Ser 20 25 30 Gly Gly Gly Gly Gly Asn Ser Thr Ala Ala Ala Ala Gly Gly Asn Gln 35 40 45 Lys Asn Ser Pro Asp Arg Val Lys Arg Pro Met Asn Ala Phe Met Val 50 55 60 Trp Ser Arg Gly Gln Arg Arg Lys Met Ala Gln Glu Asn Pro Lys Met 65 70 75 80 His Asn Ser Glu Ile Ser Lys Arg Leu Gly Ala Glu Trp Lys Leu Leu 85 90 95 Ser Glu Thr Glu Lys Arg Pro Phe Ile Asp Glu Ala Lys Arg Leu Arg 100 105 110 Ala Leu His Met Lys Glu His Pro Asp Tyr Lys Tyr Arg Pro Arg Arg 115 120 125 Lys Thr Lys Thr Leu Met Lys Lys Asp Lys Tyr Thr Leu Pro Gly Gly 130 135 140 Leu Leu Ala Pro Gly Gly Asn Ser Met Ala Ser Gly Val Gly Val Gly 145 150 155 160 Ala Gly Leu Gly Ala Gly Val Asn Gln Arg Met Asp Ser Tyr Ala His 165 170 175 Met Asn Gly Trp Ser Asn Gly Ser Tyr Ser Met Met Gln Asp Gln Leu 180 185 190 Gly Tyr Pro Gln His Pro Gly Leu Asn Ala His Gly Ala Ala Gln Met 195 200 205 Gln Pro Met His Arg Tyr Asp Val Ser Ala Leu Gln Tyr Asn Ser Met 210 215 220 Thr Ser Ser Gln Thr Tyr Met Asn Gly Ser Pro Thr Tyr Ser Met Ser 225 230 235 240 Tyr Ser Gln Gln Gly Thr Pro Gly Met Ala Leu Gly Ser Met Gly Ser 245 250 255 Val Val Lys Ser Glu Ala Ser Ser Ser Pro Pro Val Val Thr Ser Ser 260 265 270 Ser His Ser Arg Ala Pro Cys Gln Ala Gly Asp Leu Arg Asp Met Ile 275 280 285 Ser Met Tyr Leu Pro Gly Ala Glu Val Pro Glu Pro Ala Ala Pro Ser 290 295 300 Arg Leu His Met Ser Gln His Tyr Gln Ser Gly Pro Val Pro Gly Thr 305 310 315 320 Ala Ile Asn Gly Thr Leu Pro Leu Ser His Met Leu Val Ala Ala Leu 325 330 335 Leu Ala Val Leu 340 <210> 83 <211> 993 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Sox2 cDNA Sequence <400> 83 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgtacaacat gatggagacg 60 gagctgaagc cgccgggccc gcagcaaact tcggggggcg gcggcggcaa ctccaccgcg 120 gcggcggccg gcggcaacca gaaaaacagc ccggaccgcg tcaagcggcc catgaatgcc 180 ttcatggtgt ggtcccgcgg gcagcggcgc aagatggccc aggagaaccc caagatgcac 240 aactcggaga tcagcaagcg cctgggcgcc gagtggaaac ttttgtcgga gacggagaag 300 cggccgttca tcgacgaggc taagcggctg cgagcgctgc acatgaagga gcacccggat 360 tataaatacc ggccccggcg gaaaaccaag acgctcatga agaaggataa gtacacgctg 420 cccggcgggc tgctggcccc cggcggcaat agcatggcga gcggggtcgg ggtgggcgcc 480 ggcctgggcg cgggcgtgaa ccagcgcatg gacagttacg cgcacatgaa cggctggagc 540 aacggcagct acagcatgat gcaggaccag ctgggctacc cgcagcaccc gggcctcaat 600 gcgcacggcg cagcgcagat gcagcccatg caccgctacg acgtgagcgc cctgcagtac 660 aactccatga ccagctcgca gacctacatg aacggctcgc ccacctacag catgtcctac 720 tcgcagcagg gcacccctgg catggctctt ggctccatgg gttcggtggt caagtccgag 780 gccagctcca gcccccctgt ggttacctct tcctcccact ccagggcgcc ctgccaggcc 840 ggggacctcc gggacatgat cagcatgtat ctccccggcg ccgaggtgcc ggaacccgcc 900 gcccccagca gacttcacat gtcccagcac taccagagcg gcccggtgcc cggcacggcc 960 attaacggca cactgcccct ctcacacatg tga 993 <210> 84 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Sox2 Amino Acid Sequence <400> 84 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Tyr Asn 1 5 10 15 Met Met Glu Thr Glu Leu Lys Pro Pro Gly Pro Gln Gln Thr Ser Gly 20 25 30 Gly Gly Gly Gly Asn Ser Thr Ala Ala Ala Ala Gly Gly Asn Gln Lys 35 40 45 Asn Ser Pro Asp Arg Val Lys Arg Pro Met Asn Ala Phe Met Val Trp 50 55 60 Ser Arg Gly Gln Arg Arg Lys Met Ala Gln Glu Asn Pro Lys Met His 65 70 75 80 Asn Ser Glu Ile Ser Lys Arg Leu Gly Ala Glu Trp Lys Leu Leu Ser 85 90 95 Glu Thr Glu Lys Arg Pro Phe Ile Asp Glu Ala Lys Arg Leu Arg Ala 100 105 110 Leu His Met Lys Glu His Pro Asp Tyr Lys Tyr Arg Pro Arg Arg Lys 115 120 125 Thr Lys Thr Leu Met Lys Lys Asp Lys Tyr Thr Leu Pro Gly Gly Leu 130 135 140 Leu Ala Pro Gly Gly Asn Ser Met Ala Ser Gly Val Gly Val Gly Ala 145 150 155 160 Gly Leu Gly Ala Gly Val Asn Gln Arg Met Asp Ser Tyr Ala His Met 165 170 175 Asn Gly Trp Ser Asn Gly Ser Tyr Ser Met Met Gln Asp Gln Leu Gly 180 185 190 Tyr Pro Gln His Pro Gly Leu Asn Ala His Gly Ala Ala Gln Met Gln 195 200 205 Pro Met His Arg Tyr Asp Val Ser Ala Leu Gln Tyr Asn Ser Met Thr 210 215 220 Ser Ser Gln Thr Tyr Met Asn Gly Ser Pro Thr Tyr Ser Met Ser Tyr 225 230 235 240 Ser Gln Gln Gly Thr Pro Gly Met Ala Leu Gly Ser Met Gly Ser Val 245 250 255 Val Lys Ser Glu Ala Ser Ser Ser Pro Pro Val Val Thr Ser Ser Ser 260 265 270 His Ser Arg Ala Pro Cys Gln Ala Gly Asp Leu Arg Asp Met Ile Ser 275 280 285 Met Tyr Leu Pro Gly Ala Glu Val Pro Glu Pro Ala Ala Pro Ser Arg 290 295 300 Leu His Met Ser Gln His Tyr Gln Ser Gly Pro Val Pro Gly Thr Ala 305 310 315 320 Ile Asn Gly Thr Leu Pro Leu Ser His Met 325 330 <210> 85 <211> 993 <212> DNA <213> Artificial Sequence <220> <223> NLS-Sox2-JO-86 MTD cDNA Sequence <400> 85 atgaagaaga agaggaagta caacatgatg gagacggagc tgaagccgcc gggcccgcag 60 caaacttcgg ggggcggcgg cggcaactcc accgcggcgg cggccggcgg caaccagaaa 120 aacagcccgg accgcgtcaa gcggcccatg aatgccttca tggtgtggtc ccgcgggcag 180 cggcgcaaga tggcccagga gaaccccaag atgcacaact cggagatcag caagcgcctg 240 ggcgccgagt ggaaactttt gtcggagacg gagaagcggc cgttcatcga cgaggctaag 300 cggctgcgag cgctgcacat gaaggagcac ccggattata aataccggcc ccggcggaaa 360 accaagacgc tcatgaagaa ggataagtac acgctgcccg gcgggctgct ggcccccggc 420 ggcaatagca tggcgagcgg ggtcggggtg ggcgccggcc tgggcgcggg cgtgaaccag 480 cgcatggaca gttacgcgca catgaacggc tggagcaacg gcagctacag catgatgcag 540 gaccagctgg gctacccgca gcacccgggc ctcaatgcgc acggcgcagc gcagatgcag 600 cccatgcacc gctacgacgt gagcgccctg cagtacaact ccatgaccag ctcgcagacc 660 tacatgaacg gctcgcccac ctacagcatg tcctactcgc agcagggcac ccctggcatg 720 gctcttggct ccatgggttc ggtggtcaag tccgaggcca gctccagccc ccctgtggtt 780 acctcttcct cccactccag ggcgccctgc caggccgggg acctccggga catgatcagc 840 atgtatctcc ccggcgccga ggtgccggaa cccgccgccc ccagcagact tcacatgtcc 900 cagcactacc agagcggccc ggtgcccggc acggccatta acggcacact gcccctctca 960 cacatgctgg cggtgctggc ggcggcgccg tga 993 <210> 86 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> NLS-Sox2-JO-86 MTD Amino Acid Sequence <400> 86 Met Lys Lys Lys Arg Lys Tyr Asn Met Met Glu Thr Glu Leu Lys Pro 1 5 10 15 Pro Gly Pro Gln Gln Thr Ser Gly Gly Gly Gly Gly Asn Ser Thr Ala 20 25 30 Ala Ala Ala Gly Gly Asn Gln Lys Asn Ser Pro Asp Arg Val Lys Arg 35 40 45 Pro Met Asn Ala Phe Met Val Trp Ser Arg Gly Gln Arg Arg Lys Met 50 55 60 Ala Gln Glu Asn Pro Lys Met His Asn Ser Glu Ile Ser Lys Arg Leu 65 70 75 80 Gly Ala Glu Trp Lys Leu Leu Ser Glu Thr Glu Lys Arg Pro Phe Ile 85 90 95 Asp Glu Ala Lys Arg Leu Arg Ala Leu His Met Lys Glu His Pro Asp 100 105 110 Tyr Lys Tyr Arg Pro Arg Arg Lys Thr Lys Thr Leu Met Lys Lys Asp 115 120 125 Lys Tyr Thr Leu Pro Gly Gly Leu Leu Ala Pro Gly Gly Asn Ser Met 130 135 140 Ala Ser Gly Val Gly Val Gly Ala Gly Leu Gly Ala Gly Val Asn Gln 145 150 155 160 Arg Met Asp Ser Tyr Ala His Met Asn Gly Trp Ser Asn Gly Ser Tyr 165 170 175 Ser Met Met Gln Asp Gln Leu Gly Tyr Pro Gln His Pro Gly Leu Asn 180 185 190 Ala His Gly Ala Ala Gln Met Gln Pro Met His Arg Tyr Asp Val Ser 195 200 205 Ala Leu Gln Tyr Asn Ser Met Thr Ser Ser Gln Thr Tyr Met Asn Gly 210 215 220 Ser Pro Thr Tyr Ser Met Ser Tyr Ser Gln Gln Gly Thr Pro Gly Met 225 230 235 240 Ala Leu Gly Ser Met Gly Ser Val Val Lys Ser Glu Ala Ser Ser Ser 245 250 255 Pro Pro Val Val Thr Ser Ser Ser His Ser Arg Ala Pro Cys Gln Ala 260 265 270 Gly Asp Leu Arg Asp Met Ile Ser Met Tyr Leu Pro Gly Ala Glu Val 275 280 285 Pro Glu Pro Ala Ala Pro Ser Arg Leu His Met Ser Gln His Tyr Gln 290 295 300 Ser Gly Pro Val Pro Gly Thr Ala Ile Asn Gly Thr Leu Pro Leu Ser 305 310 315 320 His Met Leu Ala Val Leu Ala Ala Ala Pro 325 330 <210> 87 <211> 1017 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Sox2-JO-86 MTD cDNA Sequence <400> 87 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgtacaacat gatggagacg 60 gagctgaagc cgccgggccc gcagcaaact tcggggggcg gcggcggcaa ctccaccgcg 120 gcggcggccg gcggcaacca gaaaaacagc ccggaccgcg tcaagcggcc catgaatgcc 180 ttcatggtgt ggtcccgcgg gcagcggcgc aagatggccc aggagaaccc caagatgcac 240 aactcggaga tcagcaagcg cctgggcgcc gagtggaaac ttttgtcgga gacggagaag 300 cggccgttca tcgacgaggc taagcggctg cgagcgctgc acatgaagga gcacccggat 360 tataaatacc ggccccggcg gaaaaccaag acgctcatga agaaggataa gtacacgctg 420 cccggcgggc tgctggcccc cggcggcaat agcatggcga gcggggtcgg ggtgggcgcc 480 ggcctgggcg cgggcgtgaa ccagcgcatg gacagttacg cgcacatgaa cggctggagc 540 aacggcagct acagcatgat gcaggaccag ctgggctacc cgcagcaccc gggcctcaat 600 gcgcacggcg cagcgcagat gcagcccatg caccgctacg acgtgagcgc cctgcagtac 660 aactccatga ccagctcgca gacctacatg aacggctcgc ccacctacag catgtcctac 720 tcgcagcagg gcacccctgg catggctctt ggctccatgg gttcggtggt caagtccgag 780 gccagctcca gcccccctgt ggttacctct tcctcccact ccagggcgcc ctgccaggcc 840 ggggacctcc gggacatgat cagcatgtat ctccccggcg ccgaggtgcc ggaacccgcc 900 gcccccagca gacttcacat gtcccagcac taccagagcg gcccggtgcc cggcacggcc 960 attaacggca cactgcccct ctcacacatg ctggcggtgc tggcggcggc gccgtga 1017 <210> 88 <211> 338 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Sox2-JO-86 MTD Amino Acid Sequence <400> 88 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Tyr Asn 1 5 10 15 Met Met Glu Thr Glu Leu Lys Pro Pro Gly Pro Gln Gln Thr Ser Gly 20 25 30 Gly Gly Gly Gly Asn Ser Thr Ala Ala Ala Ala Gly Gly Asn Gln Lys 35 40 45 Asn Ser Pro Asp Arg Val Lys Arg Pro Met Asn Ala Phe Met Val Trp 50 55 60 Ser Arg Gly Gln Arg Arg Lys Met Ala Gln Glu Asn Pro Lys Met His 65 70 75 80 Asn Ser Glu Ile Ser Lys Arg Leu Gly Ala Glu Trp Lys Leu Leu Ser 85 90 95 Glu Thr Glu Lys Arg Pro Phe Ile Asp Glu Ala Lys Arg Leu Arg Ala 100 105 110 Leu His Met Lys Glu His Pro Asp Tyr Lys Tyr Arg Pro Arg Arg Lys 115 120 125 Thr Lys Thr Leu Met Lys Lys Asp Lys Tyr Thr Leu Pro Gly Gly Leu 130 135 140 Leu Ala Pro Gly Gly Asn Ser Met Ala Ser Gly Val Gly Val Gly Ala 145 150 155 160 Gly Leu Gly Ala Gly Val Asn Gln Arg Met Asp Ser Tyr Ala His Met 165 170 175 Asn Gly Trp Ser Asn Gly Ser Tyr Ser Met Met Gln Asp Gln Leu Gly 180 185 190 Tyr Pro Gln His Pro Gly Leu Asn Ala His Gly Ala Ala Gln Met Gln 195 200 205 Pro Met His Arg Tyr Asp Val Ser Ala Leu Gln Tyr Asn Ser Met Thr 210 215 220 Ser Ser Gln Thr Tyr Met Asn Gly Ser Pro Thr Tyr Ser Met Ser Tyr 225 230 235 240 Ser Gln Gln Gly Thr Pro Gly Met Ala Leu Gly Ser Met Gly Ser Val 245 250 255 Val Lys Ser Glu Ala Ser Ser Ser Pro Pro Val Val Thr Ser Ser Ser 260 265 270 His Ser Arg Ala Pro Cys Gln Ala Gly Asp Leu Arg Asp Met Ile Ser 275 280 285 Met Tyr Leu Pro Gly Ala Glu Val Pro Glu Pro Ala Ala Pro Ser Arg 290 295 300 Leu His Met Ser Gln His Tyr Gln Ser Gly Pro Val Pro Gly Thr Ala 305 310 315 320 Ile Asn Gly Thr Leu Pro Leu Ser His Met Leu Ala Val Leu Ala Ala 325 330 335 Ala Pro <210> 89 <211> 1029 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Sox2 cDNA Sequence <400> 89 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagta caacatgatg gagacggagc tgaagccgcc gggcccgcag 120 caaacttcgg ggggcggcgg cggcaactcc accgcggcgg cggccggcgg caaccagaaa 180 aacagcccgg accgcgtcaa gcggcccatg aatgccttca tggtgtggtc ccgcgggcag 240 cggcgcaaga tggcccagga gaaccccaag atgcacaact cggagatcag caagcgcctg 300 ggcgccgagt ggaaactttt gtcggagacg gagaagcggc cgttcatcga cgaggctaag 360 cggctgcgag cgctgcacat gaaggagcac ccggattata aataccggcc ccggcggaaa 420 accaagacgc tcatgaagaa ggataagtac acgctgcccg gcgggctgct ggcccccggc 480 ggcaatagca tggcgagcgg ggtcggggtg ggcgccggcc tgggcgcggg cgtgaaccag 540 cgcatggaca gttacgcgca catgaacggc tggagcaacg gcagctacag catgatgcag 600 gaccagctgg gctacccgca gcacccgggc ctcaatgcgc acggcgcagc gcagatgcag 660 cccatgcacc gctacgacgt gagcgccctg cagtacaact ccatgaccag ctcgcagacc 720 tacatgaacg gctcgcccac ctacagcatg tcctactcgc agcagggcac ccctggcatg 780 gctcttggct ccatgggttc ggtggtcaag tccgaggcca gctccagccc ccctgtggtt 840 acctcttcct cccactccag ggcgccctgc caggccgggg acctccggga catgatcagc 900 atgtatctcc ccggcgccga ggtgccggaa cccgccgccc ccagcagact tcacatgtcc 960 cagcactacc agagcggccc ggtgcccggc acggccatta acggcacact gcccctctca 1020 cacatgtga 1029 <210> 90 <211> 342 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Sox2 Amino Acid Sequence <400> 90 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Tyr Asn Met Met Glu Thr 20 25 30 Glu Leu Lys Pro Pro Gly Pro Gln Gln Thr Ser Gly Gly Gly Gly Gly 35 40 45 Asn Ser Thr Ala Ala Ala Ala Gly Gly Asn Gln Lys Asn Ser Pro Asp 50 55 60 Arg Val Lys Arg Pro Met Asn Ala Phe Met Val Trp Ser Arg Gly Gln 65 70 75 80 Arg Arg Lys Met Ala Gln Glu Asn Pro Lys Met His Asn Ser Glu Ile 85 90 95 Ser Lys Arg Leu Gly Ala Glu Trp Lys Leu Leu Ser Glu Thr Glu Lys 100 105 110 Arg Pro Phe Ile Asp Glu Ala Lys Arg Leu Arg Ala Leu His Met Lys 115 120 125 Glu His Pro Asp Tyr Lys Tyr Arg Pro Arg Arg Lys Thr Lys Thr Leu 130 135 140 Met Lys Lys Asp Lys Tyr Thr Leu Pro Gly Gly Leu Leu Ala Pro Gly 145 150 155 160 Gly Asn Ser Met Ala Ser Gly Val Gly Val Gly Ala Gly Leu Gly Ala 165 170 175 Gly Val Asn Gln Arg Met Asp Ser Tyr Ala His Met Asn Gly Trp Ser 180 185 190 Asn Gly Ser Tyr Ser Met Met Gln Asp Gln Leu Gly Tyr Pro Gln His 195 200 205 Pro Gly Leu Asn Ala His Gly Ala Ala Gln Met Gln Pro Met His Arg 210 215 220 Tyr Asp Val Ser Ala Leu Gln Tyr Asn Ser Met Thr Ser Ser Gln Thr 225 230 235 240 Tyr Met Asn Gly Ser Pro Thr Tyr Ser Met Ser Tyr Ser Gln Gln Gly 245 250 255 Thr Pro Gly Met Ala Leu Gly Ser Met Gly Ser Val Val Lys Ser Glu 260 265 270 Ala Ser Ser Ser Pro Pro Val Val Thr Ser Ser Ser His Ser Arg Ala 275 280 285 Pro Cys Gln Ala Gly Asp Leu Arg Asp Met Ile Ser Met Tyr Leu Pro 290 295 300 Gly Ala Glu Val Pro Glu Pro Ala Ala Pro Ser Arg Leu His Met Ser 305 310 315 320 Gln His Tyr Gln Ser Gly Pro Val Pro Gly Thr Ala Ile Asn Gly Thr 325 330 335 Leu Pro Leu Ser His Met 340 <210> 91 <211> 1056 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Sox2 cDNA Sequence <400> 91 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgtacaa catgatggag 120 acggagctga agccgccggg cccgcagcaa acttcggggg gcggcggcgg caactccacc 180 gcggcggcgg ccggcggcaa ccagaaaaac agcccggacc gcgtcaagcg gcccatgaat 240 gccttcatgg tgtggtcccg cgggcagcgg cgcaagatgg cccaggagaa ccccaagatg 300 cacaactcgg agatcagcaa gcgcctgggc gccgagtgga aacttttgtc ggagacggag 360 aagcggccgt tcatcgacga ggctaagcgg ctgcgagcgc tgcacatgaa ggagcacccg 420 gattataaat accggccccg gcggaaaacc aagacgctca tgaagaagga taagtacacg 480 ctgcccggcg ggctgctggc ccccggcggc aatagcatgg cgagcggggt cggggtgggc 540 gccggcctgg gcgcgggcgt gaaccagcgc atggacagtt acgcgcacat gaacggctgg 600 agcaacggca gctacagcat gatgcaggac cagctgggct acccgcagca cccgggcctc 660 aatgcgcacg gcgcagcgca gatgcagccc atgcaccgct acgacgtgag cgccctgcag 720 tacaactcca tgaccagctc gcagacctac atgaacggct cgcccaccta cagcatgtcc 780 tactcgcagc agggcacccc tggcatggct cttggctcca tgggttcggt ggtcaagtcc 840 gaggccagct ccagcccccc tgtggttacc tcttcctccc actccagggc gccctgccag 900 gccggggacc tccgggacat gatcagcatg tatctccccg gcgccgaggt gccggaaccc 960 gccgccccca gcagacttca catgtcccag cactaccaga gcggcccggt gcccggcacg 1020 gccattaacg gcacactgcc cctctcacac atgtga 1056 <210> 92 <211> 351 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Sox2 Amino Acid Sequence <400> 92 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Tyr Asn Met Met Glu Thr Glu Leu Lys Pro Pro Gly Pro 35 40 45 Gln Gln Thr Ser Gly Gly Gly Gly Gly Asn Ser Thr Ala Ala Ala Ala 50 55 60 Gly Gly Asn Gln Lys Asn Ser Pro Asp Arg Val Lys Arg Pro Met Asn 65 70 75 80 Ala Phe Met Val Trp Ser Arg Gly Gln Arg Arg Lys Met Ala Gln Glu 85 90 95 Asn Pro Lys Met His Asn Ser Glu Ile Ser Lys Arg Leu Gly Ala Glu 100 105 110 Trp Lys Leu Leu Ser Glu Thr Glu Lys Arg Pro Phe Ile Asp Glu Ala 115 120 125 Lys Arg Leu Arg Ala Leu His Met Lys Glu His Pro Asp Tyr Lys Tyr 130 135 140 Arg Pro Arg Arg Lys Thr Lys Thr Leu Met Lys Lys Asp Lys Tyr Thr 145 150 155 160 Leu Pro Gly Gly Leu Leu Ala Pro Gly Gly Asn Ser Met Ala Ser Gly 165 170 175 Val Gly Val Gly Ala Gly Leu Gly Ala Gly Val Asn Gln Arg Met Asp 180 185 190 Ser Tyr Ala His Met Asn Gly Trp Ser Asn Gly Ser Tyr Ser Met Met 195 200 205 Gln Asp Gln Leu Gly Tyr Pro Gln His Pro Gly Leu Asn Ala His Gly 210 215 220 Ala Ala Gln Met Gln Pro Met His Arg Tyr Asp Val Ser Ala Leu Gln 225 230 235 240 Tyr Asn Ser Met Thr Ser Ser Gln Thr Tyr Met Asn Gly Ser Pro Thr 245 250 255 Tyr Ser Met Ser Tyr Ser Gln Gln Gly Thr Pro Gly Met Ala Leu Gly 260 265 270 Ser Met Gly Ser Val Val Lys Ser Glu Ala Ser Ser Ser Pro Pro Val 275 280 285 Val Thr Ser Ser Ser His Ser Arg Ala Pro Cys Gln Ala Gly Asp Leu 290 295 300 Arg Asp Met Ile Ser Met Tyr Leu Pro Gly Ala Glu Val Pro Glu Pro 305 310 315 320 Ala Ala Pro Ser Arg Leu His Met Ser Gln His Tyr Gln Ser Gly Pro 325 330 335 Val Pro Gly Thr Ala Ile Asn Gly Thr Leu Pro Leu Ser His Met 340 345 350 <210> 93 <211> 1056 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Sox2-JO-84 MTD cDNA Sequence <400> 93 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagta caacatgatg gagacggagc tgaagccgcc gggcccgcag 120 caaacttcgg ggggcggcgg cggcaactcc accgcggcgg cggccggcgg caaccagaaa 180 aacagcccgg accgcgtcaa gcggcccatg aatgccttca tggtgtggtc ccgcgggcag 240 cggcgcaaga tggcccagga gaaccccaag atgcacaact cggagatcag caagcgcctg 300 ggcgccgagt ggaaactttt gtcggagacg gagaagcggc cgttcatcga cgaggctaag 360 cggctgcgag cgctgcacat gaaggagcac ccggattata aataccggcc ccggcggaaa 420 accaagacgc tcatgaagaa ggataagtac acgctgcccg gcgggctgct ggcccccggc 480 ggcaatagca tggcgagcgg ggtcggggtg ggcgccggcc tgggcgcggg cgtgaaccag 540 cgcatggaca gttacgcgca catgaacggc tggagcaacg gcagctacag catgatgcag 600 gaccagctgg gctacccgca gcacccgggc ctcaatgcgc acggcgcagc gcagatgcag 660 cccatgcacc gctacgacgt gagcgccctg cagtacaact ccatgaccag ctcgcagacc 720 tacatgaacg gctcgcccac ctacagcatg tcctactcgc agcagggcac ccctggcatg 780 gctcttggct ccatgggttc ggtggtcaag tccgaggcca gctccagccc ccctgtggtt 840 acctcttcct cccactccag ggcgccctgc caggccgggg acctccggga catgatcagc 900 atgtatctcc ccggcgccga ggtgccggaa cccgccgccc ccagcagact tcacatgtcc 960 cagcactacc agagcggccc ggtgcccggc acggccatta acggcacact gcccctctca 1020 cacatgctgg tggcggcgct gctggcggtg ctgtga 1056 <210> 94 <211> 351 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Sox2-JO-84 MTD Amino Acid Sequence <400> 94 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Tyr Asn Met Met Glu Thr 20 25 30 Glu Leu Lys Pro Pro Gly Pro Gln Gln Thr Ser Gly Gly Gly Gly Gly 35 40 45 Asn Ser Thr Ala Ala Ala Ala Gly Gly Asn Gln Lys Asn Ser Pro Asp 50 55 60 Arg Val Lys Arg Pro Met Asn Ala Phe Met Val Trp Ser Arg Gly Gln 65 70 75 80 Arg Arg Lys Met Ala Gln Glu Asn Pro Lys Met His Asn Ser Glu Ile 85 90 95 Ser Lys Arg Leu Gly Ala Glu Trp Lys Leu Leu Ser Glu Thr Glu Lys 100 105 110 Arg Pro Phe Ile Asp Glu Ala Lys Arg Leu Arg Ala Leu His Met Lys 115 120 125 Glu His Pro Asp Tyr Lys Tyr Arg Pro Arg Arg Lys Thr Lys Thr Leu 130 135 140 Met Lys Lys Asp Lys Tyr Thr Leu Pro Gly Gly Leu Leu Ala Pro Gly 145 150 155 160 Gly Asn Ser Met Ala Ser Gly Val Gly Val Gly Ala Gly Leu Gly Ala 165 170 175 Gly Val Asn Gln Arg Met Asp Ser Tyr Ala His Met Asn Gly Trp Ser 180 185 190 Asn Gly Ser Tyr Ser Met Met Gln Asp Gln Leu Gly Tyr Pro Gln His 195 200 205 Pro Gly Leu Asn Ala His Gly Ala Ala Gln Met Gln Pro Met His Arg 210 215 220 Tyr Asp Val Ser Ala Leu Gln Tyr Asn Ser Met Thr Ser Ser Gln Thr 225 230 235 240 Tyr Met Asn Gly Ser Pro Thr Tyr Ser Met Ser Tyr Ser Gln Gln Gly 245 250 255 Thr Pro Gly Met Ala Leu Gly Ser Met Gly Ser Val Val Lys Ser Glu 260 265 270 Ala Ser Ser Ser Pro Pro Val Val Thr Ser Ser Ser His Ser Arg Ala 275 280 285 Pro Cys Gln Ala Gly Asp Leu Arg Asp Met Ile Ser Met Tyr Leu Pro 290 295 300 Gly Ala Glu Val Pro Glu Pro Ala Ala Pro Ser Arg Leu His Met Ser 305 310 315 320 Gln His Tyr Gln Ser Gly Pro Val Pro Gly Thr Ala Ile Asn Gly Thr 325 330 335 Leu Pro Leu Ser His Met Leu Val Ala Ala Leu Leu Ala Val Leu 340 345 350 <210> 95 <211> 1083 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Sox2-JO-84 MTD cDNA Sequence <400> 95 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgtacaa catgatggag 120 acggagctga agccgccggg cccgcagcaa acttcggggg gcggcggcgg caactccacc 180 gcggcggcgg ccggcggcaa ccagaaaaac agcccggacc gcgtcaagcg gcccatgaat 240 gccttcatgg tgtggtcccg cgggcagcgg cgcaagatgg cccaggagaa ccccaagatg 300 cacaactcgg agatcagcaa gcgcctgggc gccgagtgga aacttttgtc ggagacggag 360 aagcggccgt tcatcgacga ggctaagcgg ctgcgagcgc tgcacatgaa ggagcacccg 420 gattataaat accggccccg gcggaaaacc aagacgctca tgaagaagga taagtacacg 480 ctgcccggcg ggctgctggc ccccggcggc aatagcatgg cgagcggggt cggggtgggc 540 gccggcctgg gcgcgggcgt gaaccagcgc atggacagtt acgcgcacat gaacggctgg 600 agcaacggca gctacagcat gatgcaggac cagctgggct acccgcagca cccgggcctc 660 aatgcgcacg gcgcagcgca gatgcagccc atgcaccgct acgacgtgag cgccctgcag 720 tacaactcca tgaccagctc gcagacctac atgaacggct cgcccaccta cagcatgtcc 780 tactcgcagc agggcacccc tggcatggct cttggctcca tgggttcggt ggtcaagtcc 840 gaggccagct ccagcccccc tgtggttacc tcttcctccc actccagggc gccctgccag 900 gccggggacc tccgggacat gatcagcatg tatctccccg gcgccgaggt gccggaaccc 960 gccgccccca gcagacttca catgtcccag cactaccaga gcggcccggt gcccggcacg 1020 gccattaacg gcacactgcc cctctcacac atgctggtgg cggcgctgct ggcggtgctg 1080 tga 1083 <210> 96 <211> 360 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Sox2-JO-84 MTD Amino Acid Sequence <400> 96 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Tyr Asn Met Met Glu Thr Glu Leu Lys Pro Pro Gly Pro 35 40 45 Gln Gln Thr Ser Gly Gly Gly Gly Gly Asn Ser Thr Ala Ala Ala Ala 50 55 60 Gly Gly Asn Gln Lys Asn Ser Pro Asp Arg Val Lys Arg Pro Met Asn 65 70 75 80 Ala Phe Met Val Trp Ser Arg Gly Gln Arg Arg Lys Met Ala Gln Glu 85 90 95 Asn Pro Lys Met His Asn Ser Glu Ile Ser Lys Arg Leu Gly Ala Glu 100 105 110 Trp Lys Leu Leu Ser Glu Thr Glu Lys Arg Pro Phe Ile Asp Glu Ala 115 120 125 Lys Arg Leu Arg Ala Leu His Met Lys Glu His Pro Asp Tyr Lys Tyr 130 135 140 Arg Pro Arg Arg Lys Thr Lys Thr Leu Met Lys Lys Asp Lys Tyr Thr 145 150 155 160 Leu Pro Gly Gly Leu Leu Ala Pro Gly Gly Asn Ser Met Ala Ser Gly 165 170 175 Val Gly Val Gly Ala Gly Leu Gly Ala Gly Val Asn Gln Arg Met Asp 180 185 190 Ser Tyr Ala His Met Asn Gly Trp Ser Asn Gly Ser Tyr Ser Met Met 195 200 205 Gln Asp Gln Leu Gly Tyr Pro Gln His Pro Gly Leu Asn Ala His Gly 210 215 220 Ala Ala Gln Met Gln Pro Met His Arg Tyr Asp Val Ser Ala Leu Gln 225 230 235 240 Tyr Asn Ser Met Thr Ser Ser Gln Thr Tyr Met Asn Gly Ser Pro Thr 245 250 255 Tyr Ser Met Ser Tyr Ser Gln Gln Gly Thr Pro Gly Met Ala Leu Gly 260 265 270 Ser Met Gly Ser Val Val Lys Ser Glu Ala Ser Ser Ser Pro Pro Val 275 280 285 Val Thr Ser Ser Ser His Ser Arg Ala Pro Cys Gln Ala Gly Asp Leu 290 295 300 Arg Asp Met Ile Ser Met Tyr Leu Pro Gly Ala Glu Val Pro Glu Pro 305 310 315 320 Ala Ala Pro Ser Arg Leu His Met Ser Gln His Tyr Gln Ser Gly Pro 325 330 335 Val Pro Gly Thr Ala Ile Asn Gly Thr Leu Pro Leu Ser His Met Leu 340 345 350 Val Ala Ala Leu Leu Ala Val Leu 355 360 <210> 97 <211> 1053 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Sox2 cDNA Sequence <400> 97 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgtacaacat gatggagacg 120 gagctgaagc cgccgggccc gcagcaaact tcggggggcg gcggcggcaa ctccaccgcg 180 gcggcggccg gcggcaacca gaaaaacagc ccggaccgcg tcaagcggcc catgaatgcc 240 ttcatggtgt ggtcccgcgg gcagcggcgc aagatggccc aggagaaccc caagatgcac 300 aactcggaga tcagcaagcg cctgggcgcc gagtggaaac ttttgtcgga gacggagaag 360 cggccgttca tcgacgaggc taagcggctg cgagcgctgc acatgaagga gcacccggat 420 tataaatacc ggccccggcg gaaaaccaag acgctcatga agaaggataa gtacacgctg 480 cccggcgggc tgctggcccc cggcggcaat agcatggcga gcggggtcgg ggtgggcgcc 540 ggcctgggcg cgggcgtgaa ccagcgcatg gacagttacg cgcacatgaa cggctggagc 600 aacggcagct acagcatgat gcaggaccag ctgggctacc cgcagcaccc gggcctcaat 660 gcgcacggcg cagcgcagat gcagcccatg caccgctacg acgtgagcgc cctgcagtac 720 aactccatga ccagctcgca gacctacatg aacggctcgc ccacctacag catgtcctac 780 tcgcagcagg gcacccctgg catggctctt ggctccatgg gttcggtggt caagtccgag 840 gccagctcca gcccccctgt ggttacctct tcctcccact ccagggcgcc ctgccaggcc 900 ggggacctcc gggacatgat cagcatgtat ctccccggcg ccgaggtgcc ggaacccgcc 960 gcccccagca gacttcacat gtcccagcac taccagagcg gcccggtgcc cggcacggcc 1020 attaacggca cactgcccct ctcacacatg tga 1053 <210> 98 <211> 350 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Sox2 Amino Acid Sequence <400> 98 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Tyr Asn Met Met Glu Thr Glu Leu Lys Pro Pro Gly Pro Gln 35 40 45 Gln Thr Ser Gly Gly Gly Gly Gly Asn Ser Thr Ala Ala Ala Ala Gly 50 55 60 Gly Asn Gln Lys Asn Ser Pro Asp Arg Val Lys Arg Pro Met Asn Ala 65 70 75 80 Phe Met Val Trp Ser Arg Gly Gln Arg Arg Lys Met Ala Gln Glu Asn 85 90 95 Pro Lys Met His Asn Ser Glu Ile Ser Lys Arg Leu Gly Ala Glu Trp 100 105 110 Lys Leu Leu Ser Glu Thr Glu Lys Arg Pro Phe Ile Asp Glu Ala Lys 115 120 125 Arg Leu Arg Ala Leu His Met Lys Glu His Pro Asp Tyr Lys Tyr Arg 130 135 140 Pro Arg Arg Lys Thr Lys Thr Leu Met Lys Lys Asp Lys Tyr Thr Leu 145 150 155 160 Pro Gly Gly Leu Leu Ala Pro Gly Gly Asn Ser Met Ala Ser Gly Val 165 170 175 Gly Val Gly Ala Gly Leu Gly Ala Gly Val Asn Gln Arg Met Asp Ser 180 185 190 Tyr Ala His Met Asn Gly Trp Ser Asn Gly Ser Tyr Ser Met Met Gln 195 200 205 Asp Gln Leu Gly Tyr Pro Gln His Pro Gly Leu Asn Ala His Gly Ala 210 215 220 Ala Gln Met Gln Pro Met His Arg Tyr Asp Val Ser Ala Leu Gln Tyr 225 230 235 240 Asn Ser Met Thr Ser Ser Gln Thr Tyr Met Asn Gly Ser Pro Thr Tyr 245 250 255 Ser Met Ser Tyr Ser Gln Gln Gly Thr Pro Gly Met Ala Leu Gly Ser 260 265 270 Met Gly Ser Val Val Lys Ser Glu Ala Ser Ser Ser Pro Pro Val Val 275 280 285 Thr Ser Ser Ser His Ser Arg Ala Pro Cys Gln Ala Gly Asp Leu Arg 290 295 300 Asp Met Ile Ser Met Tyr Leu Pro Gly Ala Glu Val Pro Glu Pro Ala 305 310 315 320 Ala Pro Ser Arg Leu His Met Ser Gln His Tyr Gln Ser Gly Pro Val 325 330 335 Pro Gly Thr Ala Ile Asn Gly Thr Leu Pro Leu Ser His Met 340 345 350 <210> 99 <211> 1053 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Sox2-JO-86 MTD cDNA Sequence <400> 99 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagta caacatgatg gagacggagc tgaagccgcc gggcccgcag 120 caaacttcgg ggggcggcgg cggcaactcc accgcggcgg cggccggcgg caaccagaaa 180 aacagcccgg accgcgtcaa gcggcccatg aatgccttca tggtgtggtc ccgcgggcag 240 cggcgcaaga tggcccagga gaaccccaag atgcacaact cggagatcag caagcgcctg 300 ggcgccgagt ggaaactttt gtcggagacg gagaagcggc cgttcatcga cgaggctaag 360 cggctgcgag cgctgcacat gaaggagcac ccggattata aataccggcc ccggcggaaa 420 accaagacgc tcatgaagaa ggataagtac acgctgcccg gcgggctgct ggcccccggc 480 ggcaatagca tggcgagcgg ggtcggggtg ggcgccggcc tgggcgcggg cgtgaaccag 540 cgcatggaca gttacgcgca catgaacggc tggagcaacg gcagctacag catgatgcag 600 gaccagctgg gctacccgca gcacccgggc ctcaatgcgc acggcgcagc gcagatgcag 660 cccatgcacc gctacgacgt gagcgccctg cagtacaact ccatgaccag ctcgcagacc 720 tacatgaacg gctcgcccac ctacagcatg tcctactcgc agcagggcac ccctggcatg 780 gctcttggct ccatgggttc ggtggtcaag tccgaggcca gctccagccc ccctgtggtt 840 acctcttcct cccactccag ggcgccctgc caggccgggg acctccggga catgatcagc 900 atgtatctcc ccggcgccga ggtgccggaa cccgccgccc ccagcagact tcacatgtcc 960 cagcactacc agagcggccc ggtgcccggc acggccatta acggcacact gcccctctca 1020 cacatgctgg cggtgctggc ggcggcgccg tga 1053 <210> 100 <211> 350 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Sox2-JO-86 MTD Amino Acid Sequence <400> 100 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Tyr Asn Met Met Glu Thr 20 25 30 Glu Leu Lys Pro Pro Gly Pro Gln Gln Thr Ser Gly Gly Gly Gly Gly 35 40 45 Asn Ser Thr Ala Ala Ala Ala Gly Gly Asn Gln Lys Asn Ser Pro Asp 50 55 60 Arg Val Lys Arg Pro Met Asn Ala Phe Met Val Trp Ser Arg Gly Gln 65 70 75 80 Arg Arg Lys Met Ala Gln Glu Asn Pro Lys Met His Asn Ser Glu Ile 85 90 95 Ser Lys Arg Leu Gly Ala Glu Trp Lys Leu Leu Ser Glu Thr Glu Lys 100 105 110 Arg Pro Phe Ile Asp Glu Ala Lys Arg Leu Arg Ala Leu His Met Lys 115 120 125 Glu His Pro Asp Tyr Lys Tyr Arg Pro Arg Arg Lys Thr Lys Thr Leu 130 135 140 Met Lys Lys Asp Lys Tyr Thr Leu Pro Gly Gly Leu Leu Ala Pro Gly 145 150 155 160 Gly Asn Ser Met Ala Ser Gly Val Gly Val Gly Ala Gly Leu Gly Ala 165 170 175 Gly Val Asn Gln Arg Met Asp Ser Tyr Ala His Met Asn Gly Trp Ser 180 185 190 Asn Gly Ser Tyr Ser Met Met Gln Asp Gln Leu Gly Tyr Pro Gln His 195 200 205 Pro Gly Leu Asn Ala His Gly Ala Ala Gln Met Gln Pro Met His Arg 210 215 220 Tyr Asp Val Ser Ala Leu Gln Tyr Asn Ser Met Thr Ser Ser Gln Thr 225 230 235 240 Tyr Met Asn Gly Ser Pro Thr Tyr Ser Met Ser Tyr Ser Gln Gln Gly 245 250 255 Thr Pro Gly Met Ala Leu Gly Ser Met Gly Ser Val Val Lys Ser Glu 260 265 270 Ala Ser Ser Ser Pro Pro Val Val Thr Ser Ser Ser His Ser Arg Ala 275 280 285 Pro Cys Gln Ala Gly Asp Leu Arg Asp Met Ile Ser Met Tyr Leu Pro 290 295 300 Gly Ala Glu Val Pro Glu Pro Ala Ala Pro Ser Arg Leu His Met Ser 305 310 315 320 Gln His Tyr Gln Ser Gly Pro Val Pro Gly Thr Ala Ile Asn Gly Thr 325 330 335 Leu Pro Leu Ser His Met Leu Ala Val Leu Ala Ala Ala Pro 340 345 350 <210> 101 <211> 1077 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Sox2-JO-86 MTD cDNA Sequence <400> 101 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgtacaacat gatggagacg 120 gagctgaagc cgccgggccc gcagcaaact tcggggggcg gcggcggcaa ctccaccgcg 180 gcggcggccg gcggcaacca gaaaaacagc ccggaccgcg tcaagcggcc catgaatgcc 240 ttcatggtgt ggtcccgcgg gcagcggcgc aagatggccc aggagaaccc caagatgcac 300 aactcggaga tcagcaagcg cctgggcgcc gagtggaaac ttttgtcgga gacggagaag 360 cggccgttca tcgacgaggc taagcggctg cgagcgctgc acatgaagga gcacccggat 420 tataaatacc ggccccggcg gaaaaccaag acgctcatga agaaggataa gtacacgctg 480 cccggcgggc tgctggcccc cggcggcaat agcatggcga gcggggtcgg ggtgggcgcc 540 ggcctgggcg cgggcgtgaa ccagcgcatg gacagttacg cgcacatgaa cggctggagc 600 aacggcagct acagcatgat gcaggaccag ctgggctacc cgcagcaccc gggcctcaat 660 gcgcacggcg cagcgcagat gcagcccatg caccgctacg acgtgagcgc cctgcagtac 720 aactccatga ccagctcgca gacctacatg aacggctcgc ccacctacag catgtcctac 780 tcgcagcagg gcacccctgg catggctctt ggctccatgg gttcggtggt caagtccgag 840 gccagctcca gcccccctgt ggttacctct tcctcccact ccagggcgcc ctgccaggcc 900 ggggacctcc gggacatgat cagcatgtat ctccccggcg ccgaggtgcc ggaacccgcc 960 gcccccagca gacttcacat gtcccagcac taccagagcg gcccggtgcc cggcacggcc 1020 attaacggca cactgcccct ctcacacatg ctggcggtgc tggcggcggc gccgtga 1077 <210> 102 <211> 358 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Sox2-JO-86 MTD Amino Acid Sequence <400> 102 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Tyr Asn Met Met Glu Thr Glu Leu Lys Pro Pro Gly Pro Gln 35 40 45 Gln Thr Ser Gly Gly Gly Gly Gly Asn Ser Thr Ala Ala Ala Ala Gly 50 55 60 Gly Asn Gln Lys Asn Ser Pro Asp Arg Val Lys Arg Pro Met Asn Ala 65 70 75 80 Phe Met Val Trp Ser Arg Gly Gln Arg Arg Lys Met Ala Gln Glu Asn 85 90 95 Pro Lys Met His Asn Ser Glu Ile Ser Lys Arg Leu Gly Ala Glu Trp 100 105 110 Lys Leu Leu Ser Glu Thr Glu Lys Arg Pro Phe Ile Asp Glu Ala Lys 115 120 125 Arg Leu Arg Ala Leu His Met Lys Glu His Pro Asp Tyr Lys Tyr Arg 130 135 140 Pro Arg Arg Lys Thr Lys Thr Leu Met Lys Lys Asp Lys Tyr Thr Leu 145 150 155 160 Pro Gly Gly Leu Leu Ala Pro Gly Gly Asn Ser Met Ala Ser Gly Val 165 170 175 Gly Val Gly Ala Gly Leu Gly Ala Gly Val Asn Gln Arg Met Asp Ser 180 185 190 Tyr Ala His Met Asn Gly Trp Ser Asn Gly Ser Tyr Ser Met Met Gln 195 200 205 Asp Gln Leu Gly Tyr Pro Gln His Pro Gly Leu Asn Ala His Gly Ala 210 215 220 Ala Gln Met Gln Pro Met His Arg Tyr Asp Val Ser Ala Leu Gln Tyr 225 230 235 240 Asn Ser Met Thr Ser Ser Gln Thr Tyr Met Asn Gly Ser Pro Thr Tyr 245 250 255 Ser Met Ser Tyr Ser Gln Gln Gly Thr Pro Gly Met Ala Leu Gly Ser 260 265 270 Met Gly Ser Val Val Lys Ser Glu Ala Ser Ser Ser Pro Pro Val Val 275 280 285 Thr Ser Ser Ser His Ser Arg Ala Pro Cys Gln Ala Gly Asp Leu Arg 290 295 300 Asp Met Ile Ser Met Tyr Leu Pro Gly Ala Glu Val Pro Glu Pro Ala 305 310 315 320 Ala Pro Ser Arg Leu His Met Ser Gln His Tyr Gln Ser Gly Pro Val 325 330 335 Pro Gly Thr Ala Ile Asn Gly Thr Leu Pro Leu Ser His Met Leu Ala 340 345 350 Val Leu Ala Ala Ala Pro 355 <210> 103 <211> 1428 <212> DNA <213> Artificial Sequence <220> <223> NLS-Klf4 cDNA Sequence <400> 103 atgaagaaga agaggaaggc tgtcagcgac gcgctgctcc catctttctc cacgttcgcg 60 tctggcccgg cgggaaggga gaagacactg cgtcaagcag gtgccccgaa taaccgctgg 120 cgggaggagc tctcccacat gaagcgactt cccccagtgc ttcccggccg cccctatgac 180 ctggcggcgg cgaccgtggc cacagacctg gagagcggcg gagccggtgc ggcttgcggc 240 ggtagcaacc tggcgcccct acctcggaga gagaccgagg agttcaacga tctcctggac 300 ctggacttta ttctctccaa ttcgctgacc catcctccgg agtcagtggc cgccaccgtg 360 tcctcgtcag cgtcagcctc ctcttcgtcg tcgccgtcga gcagcggccc tgccagcgcg 420 ccctccacct gcagcttcac ctatccgatc cgggccggga acgacccggg cgtggcgccg 480 ggcggcacgg gcggaggcct cctctatggc agggagtccg ctccccctcc gacggctccc 540 ttcaacctgg cggacatcaa cgacgtgagc ccctcgggcg gcttcgtggc cgagctcctg 600 cggccagaat tggacccggt gtacattccg ccgcagcagc cgcagccgcc aggtggcggg 660 ctgatgggca agttcgtgct gaaggcgtcg ctgagcgccc ctggcagcga gtacggcagc 720 ccgtcggtca tcagcgtcag caaaggcagc cctgacggca gccacccggt ggtggtggcg 780 ccctacaacg gcgggccgcc gcgcacgtgc cccaagatca agcaggaggc ggtctcttcg 840 tgcacccact tgggcgctgg accccctctc agcaatggcc accggccggc tgcacacgac 900 ttccccctgg ggcggcagct ccccagcagg actaccccga ccctgggtct tgaggaagtg 960 ctgagcagca gggactgtca ccctgccctg ccgcttcctc ccggcttcca tccccacccg 1020 gggcccaatt acccatcctt cctgcccgat cagatgcagc cgcaagtccc gccgctccat 1080 taccaagagc tcatgccacc cggttcctgc atgccagagg agcccaagcc aaagagggga 1140 agacgatcgt ggccccggaa aaggaccgcc acccacactt gtgattacgc gggctgcggc 1200 aaaacctaca caaagagttc ccatctcaag gcacacctgc gaacccacac aggtgagaaa 1260 ccttaccact gtgactggga cggctgtgga tggaaattcg cccgctcaga tgaactgacc 1320 aggcactacc gtaaacacac ggggcaccgc ccgttccagt gccaaaaatg cgaccgagca 1380 ttttccaggt cggaccacct cgccttacac atgaagaggc atttttaa 1428 <210> 104 <211> 475 <212> PRT <213> Artificial Sequence <220> <223> NLS-Klf4 Amino Acid Sequence <400> 104 Met Lys Lys Lys Arg Lys Ala Val Ser Asp Ala Leu Leu Pro Ser Phe 1 5 10 15 Ser Thr Phe Ala Ser Gly Pro Ala Gly Arg Glu Lys Thr Leu Arg Gln 20 25 30 Ala Gly Ala Pro Asn Asn Arg Trp Arg Glu Glu Leu Ser His Met Lys 35 40 45 Arg Leu Pro Pro Val Leu Pro Gly Arg Pro Tyr Asp Leu Ala Ala Ala 50 55 60 Thr Val Ala Thr Asp Leu Glu Ser Gly Gly Ala Gly Ala Ala Cys Gly 65 70 75 80 Gly Ser Asn Leu Ala Pro Leu Pro Arg Arg Glu Thr Glu Glu Phe Asn 85 90 95 Asp Leu Leu Asp Leu Asp Phe Ile Leu Ser Asn Ser Leu Thr His Pro 100 105 110 Pro Glu Ser Val Ala Ala Thr Val Ser Ser Ser Ala Ser Ala Ser Ser 115 120 125 Ser Ser Ser Pro Ser Ser Ser Gly Pro Ala Ser Ala Pro Ser Thr Cys 130 135 140 Ser Phe Thr Tyr Pro Ile Arg Ala Gly Asn Asp Pro Gly Val Ala Pro 145 150 155 160 Gly Gly Thr Gly Gly Gly Leu Leu Tyr Gly Arg Glu Ser Ala Pro Pro 165 170 175 Pro Thr Ala Pro Phe Asn Leu Ala Asp Ile Asn Asp Val Ser Pro Ser 180 185 190 Gly Gly Phe Val Ala Glu Leu Leu Arg Pro Glu Leu Asp Pro Val Tyr 195 200 205 Ile Pro Pro Gln Gln Pro Gln Pro Pro Gly Gly Gly Leu Met Gly Lys 210 215 220 Phe Val Leu Lys Ala Ser Leu Ser Ala Pro Gly Ser Glu Tyr Gly Ser 225 230 235 240 Pro Ser Val Ile Ser Val Ser Lys Gly Ser Pro Asp Gly Ser His Pro 245 250 255 Val Val Val Ala Pro Tyr Asn Gly Gly Pro Pro Arg Thr Cys Pro Lys 260 265 270 Ile Lys Gln Glu Ala Val Ser Ser Cys Thr His Leu Gly Ala Gly Pro 275 280 285 Pro Leu Ser Asn Gly His Arg Pro Ala Ala His Asp Phe Pro Leu Gly 290 295 300 Arg Gln Leu Pro Ser Arg Thr Thr Pro Thr Leu Gly Leu Glu Glu Val 305 310 315 320 Leu Ser Ser Arg Asp Cys His Pro Ala Leu Pro Leu Pro Pro Gly Phe 325 330 335 His Pro His Pro Gly Pro Asn Tyr Pro Ser Phe Leu Pro Asp Gln Met 340 345 350 Gln Pro Gln Val Pro Pro Leu His Tyr Gln Glu Leu Met Pro Pro Gly 355 360 365 Ser Cys Met Pro Glu Glu Pro Lys Pro Lys Arg Gly Arg Arg Ser Trp 370 375 380 Pro Arg Lys Arg Thr Ala Thr His Thr Cys Asp Tyr Ala Gly Cys Gly 385 390 395 400 Lys Thr Tyr Thr Lys Ser Ser His Leu Lys Ala His Leu Arg Thr His 405 410 415 Thr Gly Glu Lys Pro Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys 420 425 430 Phe Ala Arg Ser Asp Glu Leu Thr Arg His Tyr Arg Lys His Thr Gly 435 440 445 His Arg Pro Phe Gln Cys Gln Lys Cys Asp Arg Ala Phe Ser Arg Ser 450 455 460 Asp His Leu Ala Leu His Met Lys Arg His Phe 465 470 475 <210> 105 <211> 1455 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Klf4 cDNA Sequence <400> 105 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctggctgt cagcgacgcg 60 ctgctcccat ctttctccac gttcgcgtct ggcccggcgg gaagggagaa gacactgcgt 120 caagcaggtg ccccgaataa ccgctggcgg gaggagctct cccacatgaa gcgacttccc 180 ccagtgcttc ccggccgccc ctatgacctg gcggcggcga ccgtggccac agacctggag 240 agcggcggag ccggtgcggc ttgcggcggt agcaacctgg cgcccctacc tcggagagag 300 accgaggagt tcaacgatct cctggacctg gactttattc tctccaattc gctgacccat 360 cctccggagt cagtggccgc caccgtgtcc tcgtcagcgt cagcctcctc ttcgtcgtcg 420 ccgtcgagca gcggccctgc cagcgcgccc tccacctgca gcttcaccta tccgatccgg 480 gccgggaacg acccgggcgt ggcgccgggc ggcacgggcg gaggcctcct ctatggcagg 540 gagtccgctc cccctccgac ggctcccttc aacctggcgg acatcaacga cgtgagcccc 600 tcgggcggct tcgtggccga gctcctgcgg ccagaattgg acccggtgta cattccgccg 660 cagcagccgc agccgccagg tggcgggctg atgggcaagt tcgtgctgaa ggcgtcgctg 720 agcgcccctg gcagcgagta cggcagcccg tcggtcatca gcgtcagcaa aggcagccct 780 gacggcagcc acccggtggt ggtggcgccc tacaacggcg ggccgccgcg cacgtgcccc 840 aagatcaagc aggaggcggt ctcttcgtgc acccacttgg gcgctggacc ccctctcagc 900 aatggccacc ggccggctgc acacgacttc cccctggggc ggcagctccc cagcaggact 960 accccgaccc tgggtcttga ggaagtgctg agcagcaggg actgtcaccc tgccctgccg 1020 cttcctcccg gcttccatcc ccacccgggg cccaattacc catccttcct gcccgatcag 1080 atgcagccgc aagtcccgcc gctccattac caagagctca tgccacccgg ttcctgcatg 1140 ccagaggagc ccaagccaaa gaggggaaga cgatcgtggc cccggaaaag gaccgccacc 1200 cacacttgtg attacgcggg ctgcggcaaa acctacacaa agagttccca tctcaaggca 1260 cacctgcgaa cccacacagg tgagaaacct taccactgtg actgggacgg ctgtggatgg 1320 aaattcgccc gctcagatga actgaccagg cactaccgta aacacacggg gcaccgcccg 1380 ttccagtgcc aaaaatgcga ccgagcattt tccaggtcgg accacctcgc cttacacatg 1440 aagaggcatt tttaa 1455 <210> 106 <211> 484 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Klf4 Amino Acid Sequence <400> 106 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Ala 1 5 10 15 Val Ser Asp Ala Leu Leu Pro Ser Phe Ser Thr Phe Ala Ser Gly Pro 20 25 30 Ala Gly Arg Glu Lys Thr Leu Arg Gln Ala Gly Ala Pro Asn Asn Arg 35 40 45 Trp Arg Glu Glu Leu Ser His Met Lys Arg Leu Pro Pro Val Leu Pro 50 55 60 Gly Arg Pro Tyr Asp Leu Ala Ala Ala Thr Val Ala Thr Asp Leu Glu 65 70 75 80 Ser Gly Gly Ala Gly Ala Ala Cys Gly Gly Ser Asn Leu Ala Pro Leu 85 90 95 Pro Arg Arg Glu Thr Glu Glu Phe Asn Asp Leu Leu Asp Leu Asp Phe 100 105 110 Ile Leu Ser Asn Ser Leu Thr His Pro Pro Glu Ser Val Ala Ala Thr 115 120 125 Val Ser Ser Ser Ala Ser Ala Ser Ser Ser Ser Ser Pro Ser Ser Ser 130 135 140 Gly Pro Ala Ser Ala Pro Ser Thr Cys Ser Phe Thr Tyr Pro Ile Arg 145 150 155 160 Ala Gly Asn Asp Pro Gly Val Ala Pro Gly Gly Thr Gly Gly Gly Leu 165 170 175 Leu Tyr Gly Arg Glu Ser Ala Pro Pro Pro Thr Ala Pro Phe Asn Leu 180 185 190 Ala Asp Ile Asn Asp Val Ser Pro Ser Gly Gly Phe Val Ala Glu Leu 195 200 205 Leu Arg Pro Glu Leu Asp Pro Val Tyr Ile Pro Pro Gln Gln Pro Gln 210 215 220 Pro Pro Gly Gly Gly Leu Met Gly Lys Phe Val Leu Lys Ala Ser Leu 225 230 235 240 Ser Ala Pro Gly Ser Glu Tyr Gly Ser Pro Ser Val Ile Ser Val Ser 245 250 255 Lys Gly Ser Pro Asp Gly Ser His Pro Val Val Val Ala Pro Tyr Asn 260 265 270 Gly Gly Pro Pro Arg Thr Cys Pro Lys Ile Lys Gln Glu Ala Val Ser 275 280 285 Ser Cys Thr His Leu Gly Ala Gly Pro Pro Leu Ser Asn Gly His Arg 290 295 300 Pro Ala Ala His Asp Phe Pro Leu Gly Arg Gln Leu Pro Ser Arg Thr 305 310 315 320 Thr Pro Thr Leu Gly Leu Glu Glu Val Leu Ser Ser Arg Asp Cys His 325 330 335 Pro Ala Leu Pro Leu Pro Pro Gly Phe His Pro His Pro Gly Pro Asn 340 345 350 Tyr Pro Ser Phe Leu Pro Asp Gln Met Gln Pro Gln Val Pro Pro Leu 355 360 365 His Tyr Gln Glu Leu Met Pro Pro Gly Ser Cys Met Pro Glu Glu Pro 370 375 380 Lys Pro Lys Arg Gly Arg Arg Ser Trp Pro Arg Lys Arg Thr Ala Thr 385 390 395 400 His Thr Cys Asp Tyr Ala Gly Cys Gly Lys Thr Tyr Thr Lys Ser Ser 405 410 415 His Leu Lys Ala His Leu Arg Thr His Thr Gly Glu Lys Pro Tyr His 420 425 430 Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu Leu 435 440 445 Thr Arg His Tyr Arg Lys His Thr Gly His Arg Pro Phe Gln Cys Gln 450 455 460 Lys Cys Asp Arg Ala Phe Ser Arg Ser Asp His Leu Ala Leu His Met 465 470 475 480 Lys Arg His Phe <210> 107 <211> 1455 <212> DNA <213> Artificial Sequence <220> <223> NLS-Klf4-JO-84 MTD cDNA Sequence <400> 107 atgaagaaga agaggaaggc tgtcagcgac gcgctgctcc catctttctc cacgttcgcg 60 tctggcccgg cgggaaggga gaagacactg cgtcaagcag gtgccccgaa taaccgctgg 120 cgggaggagc tctcccacat gaagcgactt cccccagtgc ttcccggccg cccctatgac 180 ctggcggcgg cgaccgtggc cacagacctg gagagcggcg gagccggtgc ggcttgcggc 240 ggtagcaacc tggcgcccct acctcggaga gagaccgagg agttcaacga tctcctggac 300 ctggacttta ttctctccaa ttcgctgacc catcctccgg agtcagtggc cgccaccgtg 360 tcctcgtcag cgtcagcctc ctcttcgtcg tcgccgtcga gcagcggccc tgccagcgcg 420 ccctccacct gcagcttcac ctatccgatc cgggccggga acgacccggg cgtggcgccg 480 ggcggcacgg gcggaggcct cctctatggc agggagtccg ctccccctcc gacggctccc 540 ttcaacctgg cggacatcaa cgacgtgagc ccctcgggcg gcttcgtggc cgagctcctg 600 cggccagaat tggacccggt gtacattccg ccgcagcagc cgcagccgcc aggtggcggg 660 ctgatgggca agttcgtgct gaaggcgtcg ctgagcgccc ctggcagcga gtacggcagc 720 ccgtcggtca tcagcgtcag caaaggcagc cctgacggca gccacccggt ggtggtggcg 780 ccctacaacg gcgggccgcc gcgcacgtgc cccaagatca agcaggaggc ggtctcttcg 840 tgcacccact tgggcgctgg accccctctc agcaatggcc accggccggc tgcacacgac 900 ttccccctgg ggcggcagct ccccagcagg actaccccga ccctgggtct tgaggaagtg 960 ctgagcagca gggactgtca ccctgccctg ccgcttcctc ccggcttcca tccccacccg 1020 gggcccaatt acccatcctt cctgcccgat cagatgcagc cgcaagtccc gccgctccat 1080 taccaagagc tcatgccacc cggttcctgc atgccagagg agcccaagcc aaagagggga 1140 agacgatcgt ggccccggaa aaggaccgcc acccacactt gtgattacgc gggctgcggc 1200 aaaacctaca caaagagttc ccatctcaag gcacacctgc gaacccacac aggtgagaaa 1260 ccttaccact gtgactggga cggctgtgga tggaaattcg cccgctcaga tgaactgacc 1320 aggcactacc gtaaacacac ggggcaccgc ccgttccagt gccaaaaatg cgaccgagca 1380 ttttccaggt cggaccacct cgccttacac atgaagaggc attttctggt ggcggcgctg 1440 ctggcggtgc tgtaa 1455 <210> 108 <211> 484 <212> PRT <213> Artificial Sequence <220> <223> NLS-Klf4-JO-84 MTD Amino Acid Sequence <400> 108 Met Lys Lys Lys Arg Lys Ala Val Ser Asp Ala Leu Leu Pro Ser Phe 1 5 10 15 Ser Thr Phe Ala Ser Gly Pro Ala Gly Arg Glu Lys Thr Leu Arg Gln 20 25 30 Ala Gly Ala Pro Asn Asn Arg Trp Arg Glu Glu Leu Ser His Met Lys 35 40 45 Arg Leu Pro Pro Val Leu Pro Gly Arg Pro Tyr Asp Leu Ala Ala Ala 50 55 60 Thr Val Ala Thr Asp Leu Glu Ser Gly Gly Ala Gly Ala Ala Cys Gly 65 70 75 80 Gly Ser Asn Leu Ala Pro Leu Pro Arg Arg Glu Thr Glu Glu Phe Asn 85 90 95 Asp Leu Leu Asp Leu Asp Phe Ile Leu Ser Asn Ser Leu Thr His Pro 100 105 110 Pro Glu Ser Val Ala Ala Thr Val Ser Ser Ser Ala Ser Ala Ser Ser 115 120 125 Ser Ser Ser Pro Ser Ser Ser Gly Pro Ala Ser Ala Pro Ser Thr Cys 130 135 140 Ser Phe Thr Tyr Pro Ile Arg Ala Gly Asn Asp Pro Gly Val Ala Pro 145 150 155 160 Gly Gly Thr Gly Gly Gly Leu Leu Tyr Gly Arg Glu Ser Ala Pro Pro 165 170 175 Pro Thr Ala Pro Phe Asn Leu Ala Asp Ile Asn Asp Val Ser Pro Ser 180 185 190 Gly Gly Phe Val Ala Glu Leu Leu Arg Pro Glu Leu Asp Pro Val Tyr 195 200 205 Ile Pro Pro Gln Gln Pro Gln Pro Pro Gly Gly Gly Leu Met Gly Lys 210 215 220 Phe Val Leu Lys Ala Ser Leu Ser Ala Pro Gly Ser Glu Tyr Gly Ser 225 230 235 240 Pro Ser Val Ile Ser Val Ser Lys Gly Ser Pro Asp Gly Ser His Pro 245 250 255 Val Val Val Ala Pro Tyr Asn Gly Gly Pro Pro Arg Thr Cys Pro Lys 260 265 270 Ile Lys Gln Glu Ala Val Ser Ser Cys Thr His Leu Gly Ala Gly Pro 275 280 285 Pro Leu Ser Asn Gly His Arg Pro Ala Ala His Asp Phe Pro Leu Gly 290 295 300 Arg Gln Leu Pro Ser Arg Thr Thr Pro Thr Leu Gly Leu Glu Glu Val 305 310 315 320 Leu Ser Ser Arg Asp Cys His Pro Ala Leu Pro Leu Pro Pro Gly Phe 325 330 335 His Pro His Pro Gly Pro Asn Tyr Pro Ser Phe Leu Pro Asp Gln Met 340 345 350 Gln Pro Gln Val Pro Pro Leu His Tyr Gln Glu Leu Met Pro Pro Gly 355 360 365 Ser Cys Met Pro Glu Glu Pro Lys Pro Lys Arg Gly Arg Arg Ser Trp 370 375 380 Pro Arg Lys Arg Thr Ala Thr His Thr Cys Asp Tyr Ala Gly Cys Gly 385 390 395 400 Lys Thr Tyr Thr Lys Ser Ser His Leu Lys Ala His Leu Arg Thr His 405 410 415 Thr Gly Glu Lys Pro Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys 420 425 430 Phe Ala Arg Ser Asp Glu Leu Thr Arg His Tyr Arg Lys His Thr Gly 435 440 445 His Arg Pro Phe Gln Cys Gln Lys Cys Asp Arg Ala Phe Ser Arg Ser 450 455 460 Asp His Leu Ala Leu His Met Lys Arg His Phe Leu Val Ala Ala Leu 465 470 475 480 Leu Ala Val Leu <210> 109 <211> 1482 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Klf4-JO-84 MTD cDNA Sequence <400> 109 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctggctgt cagcgacgcg 60 ctgctcccat ctttctccac gttcgcgtct ggcccggcgg gaagggagaa gacactgcgt 120 caagcaggtg ccccgaataa ccgctggcgg gaggagctct cccacatgaa gcgacttccc 180 ccagtgcttc ccggccgccc ctatgacctg gcggcggcga ccgtggccac agacctggag 240 agcggcggag ccggtgcggc ttgcggcggt agcaacctgg cgcccctacc tcggagagag 300 accgaggagt tcaacgatct cctggacctg gactttattc tctccaattc gctgacccat 360 cctccggagt cagtggccgc caccgtgtcc tcgtcagcgt cagcctcctc ttcgtcgtcg 420 ccgtcgagca gcggccctgc cagcgcgccc tccacctgca gcttcaccta tccgatccgg 480 gccgggaacg acccgggcgt ggcgccgggc ggcacgggcg gaggcctcct ctatggcagg 540 gagtccgctc cccctccgac ggctcccttc aacctggcgg acatcaacga cgtgagcccc 600 tcgggcggct tcgtggccga gctcctgcgg ccagaattgg acccggtgta cattccgccg 660 cagcagccgc agccgccagg tggcgggctg atgggcaagt tcgtgctgaa ggcgtcgctg 720 agcgcccctg gcagcgagta cggcagcccg tcggtcatca gcgtcagcaa aggcagccct 780 gacggcagcc acccggtggt ggtggcgccc tacaacggcg ggccgccgcg cacgtgcccc 840 aagatcaagc aggaggcggt ctcttcgtgc acccacttgg gcgctggacc ccctctcagc 900 aatggccacc ggccggctgc acacgacttc cccctggggc ggcagctccc cagcaggact 960 accccgaccc tgggtcttga ggaagtgctg agcagcaggg actgtcaccc tgccctgccg 1020 cttcctcccg gcttccatcc ccacccgggg cccaattacc catccttcct gcccgatcag 1080 atgcagccgc aagtcccgcc gctccattac caagagctca tgccacccgg ttcctgcatg 1140 ccagaggagc ccaagccaaa gaggggaaga cgatcgtggc cccggaaaag gaccgccacc 1200 cacacttgtg attacgcggg ctgcggcaaa acctacacaa agagttccca tctcaaggca 1260 cacctgcgaa cccacacagg tgagaaacct taccactgtg actgggacgg ctgtggatgg 1320 aaattcgccc gctcagatga actgaccagg cactaccgta aacacacggg gcaccgcccg 1380 ttccagtgcc aaaaatgcga ccgagcattt tccaggtcgg accacctcgc cttacacatg 1440 aagaggcatt ttctggtggc ggcgctgctg gcggtgctgt aa 1482 <210> 110 <211> 493 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Klf4-JO-84 MTD Amino Acid Sequence <400> 110 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Ala 1 5 10 15 Val Ser Asp Ala Leu Leu Pro Ser Phe Ser Thr Phe Ala Ser Gly Pro 20 25 30 Ala Gly Arg Glu Lys Thr Leu Arg Gln Ala Gly Ala Pro Asn Asn Arg 35 40 45 Trp Arg Glu Glu Leu Ser His Met Lys Arg Leu Pro Pro Val Leu Pro 50 55 60 Gly Arg Pro Tyr Asp Leu Ala Ala Ala Thr Val Ala Thr Asp Leu Glu 65 70 75 80 Ser Gly Gly Ala Gly Ala Ala Cys Gly Gly Ser Asn Leu Ala Pro Leu 85 90 95 Pro Arg Arg Glu Thr Glu Glu Phe Asn Asp Leu Leu Asp Leu Asp Phe 100 105 110 Ile Leu Ser Asn Ser Leu Thr His Pro Pro Glu Ser Val Ala Ala Thr 115 120 125 Val Ser Ser Ser Ala Ser Ala Ser Ser Ser Ser Ser Pro Ser Ser Ser 130 135 140 Gly Pro Ala Ser Ala Pro Ser Thr Cys Ser Phe Thr Tyr Pro Ile Arg 145 150 155 160 Ala Gly Asn Asp Pro Gly Val Ala Pro Gly Gly Thr Gly Gly Gly Leu 165 170 175 Leu Tyr Gly Arg Glu Ser Ala Pro Pro Pro Thr Ala Pro Phe Asn Leu 180 185 190 Ala Asp Ile Asn Asp Val Ser Pro Ser Gly Gly Phe Val Ala Glu Leu 195 200 205 Leu Arg Pro Glu Leu Asp Pro Val Tyr Ile Pro Pro Gln Gln Pro Gln 210 215 220 Pro Pro Gly Gly Gly Leu Met Gly Lys Phe Val Leu Lys Ala Ser Leu 225 230 235 240 Ser Ala Pro Gly Ser Glu Tyr Gly Ser Pro Ser Val Ile Ser Val Ser 245 250 255 Lys Gly Ser Pro Asp Gly Ser His Pro Val Val Val Ala Pro Tyr Asn 260 265 270 Gly Gly Pro Pro Arg Thr Cys Pro Lys Ile Lys Gln Glu Ala Val Ser 275 280 285 Ser Cys Thr His Leu Gly Ala Gly Pro Pro Leu Ser Asn Gly His Arg 290 295 300 Pro Ala Ala His Asp Phe Pro Leu Gly Arg Gln Leu Pro Ser Arg Thr 305 310 315 320 Thr Pro Thr Leu Gly Leu Glu Glu Val Leu Ser Ser Arg Asp Cys His 325 330 335 Pro Ala Leu Pro Leu Pro Pro Gly Phe His Pro His Pro Gly Pro Asn 340 345 350 Tyr Pro Ser Phe Leu Pro Asp Gln Met Gln Pro Gln Val Pro Pro Leu 355 360 365 His Tyr Gln Glu Leu Met Pro Pro Gly Ser Cys Met Pro Glu Glu Pro 370 375 380 Lys Pro Lys Arg Gly Arg Arg Ser Trp Pro Arg Lys Arg Thr Ala Thr 385 390 395 400 His Thr Cys Asp Tyr Ala Gly Cys Gly Lys Thr Tyr Thr Lys Ser Ser 405 410 415 His Leu Lys Ala His Leu Arg Thr His Thr Gly Glu Lys Pro Tyr His 420 425 430 Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu Leu 435 440 445 Thr Arg His Tyr Arg Lys His Thr Gly His Arg Pro Phe Gln Cys Gln 450 455 460 Lys Cys Asp Arg Ala Phe Ser Arg Ser Asp His Leu Ala Leu His Met 465 470 475 480 Lys Arg His Phe Leu Val Ala Ala Leu Leu Ala Val Leu 485 490 <210> 111 <211> 1452 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Klf4 cDNA Sequence <400> 111 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cggctgtcag cgacgcgctg 60 ctcccatctt tctccacgtt cgcgtctggc ccggcgggaa gggagaagac actgcgtcaa 120 gcaggtgccc cgaataaccg ctggcgggag gagctctccc acatgaagcg acttccccca 180 gtgcttcccg gccgccccta tgacctggcg gcggcgaccg tggccacaga cctggagagc 240 ggcggagccg gtgcggcttg cggcggtagc aacctggcgc ccctacctcg gagagagacc 300 gaggagttca acgatctcct ggacctggac tttattctct ccaattcgct gacccatcct 360 ccggagtcag tggccgccac cgtgtcctcg tcagcgtcag cctcctcttc gtcgtcgccg 420 tcgagcagcg gccctgccag cgcgccctcc acctgcagct tcacctatcc gatccgggcc 480 gggaacgacc cgggcgtggc gccgggcggc acgggcggag gcctcctcta tggcagggag 540 tccgctcccc ctccgacggc tcccttcaac ctggcggaca tcaacgacgt gagcccctcg 600 ggcggcttcg tggccgagct cctgcggcca gaattggacc cggtgtacat tccgccgcag 660 cagccgcagc cgccaggtgg cgggctgatg ggcaagttcg tgctgaaggc gtcgctgagc 720 gcccctggca gcgagtacgg cagcccgtcg gtcatcagcg tcagcaaagg cagccctgac 780 ggcagccacc cggtggtggt ggcgccctac aacggcgggc cgccgcgcac gtgccccaag 840 atcaagcagg aggcggtctc ttcgtgcacc cacttgggcg ctggaccccc tctcagcaat 900 ggccaccggc cggctgcaca cgacttcccc ctggggcggc agctccccag caggactacc 960 ccgaccctgg gtcttgagga agtgctgagc agcagggact gtcaccctgc cctgccgctt 1020 cctcccggct tccatcccca cccggggccc aattacccat ccttcctgcc cgatcagatg 1080 cagccgcaag tcccgccgct ccattaccaa gagctcatgc cacccggttc ctgcatgcca 1140 gaggagccca agccaaagag gggaagacga tcgtggcccc ggaaaaggac cgccacccac 1200 acttgtgatt acgcgggctg cggcaaaacc tacacaaaga gttcccatct caaggcacac 1260 ctgcgaaccc acacaggtga gaaaccttac cactgtgact gggacggctg tggatggaaa 1320 ttcgcccgct cagatgaact gaccaggcac taccgtaaac acacggggca ccgcccgttc 1380 cagtgccaaa aatgcgaccg agcattttcc aggtcggacc acctcgcctt acacatgaag 1440 aggcattttt aa 1452 <210> 112 <211> 483 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Klf4 Amino Acid Sequence <400> 112 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Ala Val 1 5 10 15 Ser Asp Ala Leu Leu Pro Ser Phe Ser Thr Phe Ala Ser Gly Pro Ala 20 25 30 Gly Arg Glu Lys Thr Leu Arg Gln Ala Gly Ala Pro Asn Asn Arg Trp 35 40 45 Arg Glu Glu Leu Ser His Met Lys Arg Leu Pro Pro Val Leu Pro Gly 50 55 60 Arg Pro Tyr Asp Leu Ala Ala Ala Thr Val Ala Thr Asp Leu Glu Ser 65 70 75 80 Gly Gly Ala Gly Ala Ala Cys Gly Gly Ser Asn Leu Ala Pro Leu Pro 85 90 95 Arg Arg Glu Thr Glu Glu Phe Asn Asp Leu Leu Asp Leu Asp Phe Ile 100 105 110 Leu Ser Asn Ser Leu Thr His Pro Pro Glu Ser Val Ala Ala Thr Val 115 120 125 Ser Ser Ser Ala Ser Ala Ser Ser Ser Ser Ser Pro Ser Ser Ser Gly 130 135 140 Pro Ala Ser Ala Pro Ser Thr Cys Ser Phe Thr Tyr Pro Ile Arg Ala 145 150 155 160 Gly Asn Asp Pro Gly Val Ala Pro Gly Gly Thr Gly Gly Gly Leu Leu 165 170 175 Tyr Gly Arg Glu Ser Ala Pro Pro Pro Thr Ala Pro Phe Asn Leu Ala 180 185 190 Asp Ile Asn Asp Val Ser Pro Ser Gly Gly Phe Val Ala Glu Leu Leu 195 200 205 Arg Pro Glu Leu Asp Pro Val Tyr Ile Pro Pro Gln Gln Pro Gln Pro 210 215 220 Pro Gly Gly Gly Leu Met Gly Lys Phe Val Leu Lys Ala Ser Leu Ser 225 230 235 240 Ala Pro Gly Ser Glu Tyr Gly Ser Pro Ser Val Ile Ser Val Ser Lys 245 250 255 Gly Ser Pro Asp Gly Ser His Pro Val Val Val Ala Pro Tyr Asn Gly 260 265 270 Gly Pro Pro Arg Thr Cys Pro Lys Ile Lys Gln Glu Ala Val Ser Ser 275 280 285 Cys Thr His Leu Gly Ala Gly Pro Pro Leu Ser Asn Gly His Arg Pro 290 295 300 Ala Ala His Asp Phe Pro Leu Gly Arg Gln Leu Pro Ser Arg Thr Thr 305 310 315 320 Pro Thr Leu Gly Leu Glu Glu Val Leu Ser Ser Arg Asp Cys His Pro 325 330 335 Ala Leu Pro Leu Pro Pro Gly Phe His Pro His Pro Gly Pro Asn Tyr 340 345 350 Pro Ser Phe Leu Pro Asp Gln Met Gln Pro Gln Val Pro Pro Leu His 355 360 365 Tyr Gln Glu Leu Met Pro Pro Gly Ser Cys Met Pro Glu Glu Pro Lys 370 375 380 Pro Lys Arg Gly Arg Arg Ser Trp Pro Arg Lys Arg Thr Ala Thr His 385 390 395 400 Thr Cys Asp Tyr Ala Gly Cys Gly Lys Thr Tyr Thr Lys Ser Ser His 405 410 415 Leu Lys Ala His Leu Arg Thr His Thr Gly Glu Lys Pro Tyr His Cys 420 425 430 Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu Leu Thr 435 440 445 Arg His Tyr Arg Lys His Thr Gly His Arg Pro Phe Gln Cys Gln Lys 450 455 460 Cys Asp Arg Ala Phe Ser Arg Ser Asp His Leu Ala Leu His Met Lys 465 470 475 480 Arg His Phe <210> 113 <211> 1452 <212> DNA <213> Artificial Sequence <220> <223> NLS-Klf4-JO-86 MTD cDNA Sequence <400> 113 atgaagaaga agaggaaggc tgtcagcgac gcgctgctcc catctttctc cacgttcgcg 60 tctggcccgg cgggaaggga gaagacactg cgtcaagcag gtgccccgaa taaccgctgg 120 cgggaggagc tctcccacat gaagcgactt cccccagtgc ttcccggccg cccctatgac 180 ctggcggcgg cgaccgtggc cacagacctg gagagcggcg gagccggtgc ggcttgcggc 240 ggtagcaacc tggcgcccct acctcggaga gagaccgagg agttcaacga tctcctggac 300 ctggacttta ttctctccaa ttcgctgacc catcctccgg agtcagtggc cgccaccgtg 360 tcctcgtcag cgtcagcctc ctcttcgtcg tcgccgtcga gcagcggccc tgccagcgcg 420 ccctccacct gcagcttcac ctatccgatc cgggccggga acgacccggg cgtggcgccg 480 ggcggcacgg gcggaggcct cctctatggc agggagtccg ctccccctcc gacggctccc 540 ttcaacctgg cggacatcaa cgacgtgagc ccctcgggcg gcttcgtggc cgagctcctg 600 cggccagaat tggacccggt gtacattccg ccgcagcagc cgcagccgcc aggtggcggg 660 ctgatgggca agttcgtgct gaaggcgtcg ctgagcgccc ctggcagcga gtacggcagc 720 ccgtcggtca tcagcgtcag caaaggcagc cctgacggca gccacccggt ggtggtggcg 780 ccctacaacg gcgggccgcc gcgcacgtgc cccaagatca agcaggaggc ggtctcttcg 840 tgcacccact tgggcgctgg accccctctc agcaatggcc accggccggc tgcacacgac 900 ttccccctgg ggcggcagct ccccagcagg actaccccga ccctgggtct tgaggaagtg 960 ctgagcagca gggactgtca ccctgccctg ccgcttcctc ccggcttcca tccccacccg 1020 gggcccaatt acccatcctt cctgcccgat cagatgcagc cgcaagtccc gccgctccat 1080 taccaagagc tcatgccacc cggttcctgc atgccagagg agcccaagcc aaagagggga 1140 agacgatcgt ggccccggaa aaggaccgcc acccacactt gtgattacgc gggctgcggc 1200 aaaacctaca caaagagttc ccatctcaag gcacacctgc gaacccacac aggtgagaaa 1260 ccttaccact gtgactggga cggctgtgga tggaaattcg cccgctcaga tgaactgacc 1320 aggcactacc gtaaacacac ggggcaccgc ccgttccagt gccaaaaatg cgaccgagca 1380 ttttccaggt cggaccacct cgccttacac atgaagaggc attttctggc ggtgctggcg 1440 gcggcgccgt aa 1452 <210> 114 <211> 483 <212> PRT <213> Artificial Sequence <220> <223> NLS-Klf4-JO-86 MTD Amino Acid Sequence <400> 114 Met Lys Lys Lys Arg Lys Ala Val Ser Asp Ala Leu Leu Pro Ser Phe 1 5 10 15 Ser Thr Phe Ala Ser Gly Pro Ala Gly Arg Glu Lys Thr Leu Arg Gln 20 25 30 Ala Gly Ala Pro Asn Asn Arg Trp Arg Glu Glu Leu Ser His Met Lys 35 40 45 Arg Leu Pro Pro Val Leu Pro Gly Arg Pro Tyr Asp Leu Ala Ala Ala 50 55 60 Thr Val Ala Thr Asp Leu Glu Ser Gly Gly Ala Gly Ala Ala Cys Gly 65 70 75 80 Gly Ser Asn Leu Ala Pro Leu Pro Arg Arg Glu Thr Glu Glu Phe Asn 85 90 95 Asp Leu Leu Asp Leu Asp Phe Ile Leu Ser Asn Ser Leu Thr His Pro 100 105 110 Pro Glu Ser Val Ala Ala Thr Val Ser Ser Ser Ala Ser Ala Ser Ser 115 120 125 Ser Ser Ser Pro Ser Ser Ser Gly Pro Ala Ser Ala Pro Ser Thr Cys 130 135 140 Ser Phe Thr Tyr Pro Ile Arg Ala Gly Asn Asp Pro Gly Val Ala Pro 145 150 155 160 Gly Gly Thr Gly Gly Gly Leu Leu Tyr Gly Arg Glu Ser Ala Pro Pro 165 170 175 Pro Thr Ala Pro Phe Asn Leu Ala Asp Ile Asn Asp Val Ser Pro Ser 180 185 190 Gly Gly Phe Val Ala Glu Leu Leu Arg Pro Glu Leu Asp Pro Val Tyr 195 200 205 Ile Pro Pro Gln Gln Pro Gln Pro Pro Gly Gly Gly Leu Met Gly Lys 210 215 220 Phe Val Leu Lys Ala Ser Leu Ser Ala Pro Gly Ser Glu Tyr Gly Ser 225 230 235 240 Pro Ser Val Ile Ser Val Ser Lys Gly Ser Pro Asp Gly Ser His Pro 245 250 255 Val Val Val Ala Pro Tyr Asn Gly Gly Pro Pro Arg Thr Cys Pro Lys 260 265 270 Ile Lys Gln Glu Ala Val Ser Ser Cys Thr His Leu Gly Ala Gly Pro 275 280 285 Pro Leu Ser Asn Gly His Arg Pro Ala Ala His Asp Phe Pro Leu Gly 290 295 300 Arg Gln Leu Pro Ser Arg Thr Thr Pro Thr Leu Gly Leu Glu Glu Val 305 310 315 320 Leu Ser Ser Arg Asp Cys His Pro Ala Leu Pro Leu Pro Pro Gly Phe 325 330 335 His Pro His Pro Gly Pro Asn Tyr Pro Ser Phe Leu Pro Asp Gln Met 340 345 350 Gln Pro Gln Val Pro Pro Leu His Tyr Gln Glu Leu Met Pro Pro Gly 355 360 365 Ser Cys Met Pro Glu Glu Pro Lys Pro Lys Arg Gly Arg Arg Ser Trp 370 375 380 Pro Arg Lys Arg Thr Ala Thr His Thr Cys Asp Tyr Ala Gly Cys Gly 385 390 395 400 Lys Thr Tyr Thr Lys Ser Ser His Leu Lys Ala His Leu Arg Thr His 405 410 415 Thr Gly Glu Lys Pro Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys 420 425 430 Phe Ala Arg Ser Asp Glu Leu Thr Arg His Tyr Arg Lys His Thr Gly 435 440 445 His Arg Pro Phe Gln Cys Gln Lys Cys Asp Arg Ala Phe Ser Arg Ser 450 455 460 Asp His Leu Ala Leu His Met Lys Arg His Phe Leu Ala Val Leu Ala 465 470 475 480 Ala Ala Pro <210> 115 <211> 1476 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Klf4-JO-86 MTD cDNA Sequence <400> 115 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cggctgtcag cgacgcgctg 60 ctcccatctt tctccacgtt cgcgtctggc ccggcgggaa gggagaagac actgcgtcaa 120 gcaggtgccc cgaataaccg ctggcgggag gagctctccc acatgaagcg acttccccca 180 gtgcttcccg gccgccccta tgacctggcg gcggcgaccg tggccacaga cctggagagc 240 ggcggagccg gtgcggcttg cggcggtagc aacctggcgc ccctacctcg gagagagacc 300 gaggagttca acgatctcct ggacctggac tttattctct ccaattcgct gacccatcct 360 ccggagtcag tggccgccac cgtgtcctcg tcagcgtcag cctcctcttc gtcgtcgccg 420 tcgagcagcg gccctgccag cgcgccctcc acctgcagct tcacctatcc gatccgggcc 480 gggaacgacc cgggcgtggc gccgggcggc acgggcggag gcctcctcta tggcagggag 540 tccgctcccc ctccgacggc tcccttcaac ctggcggaca tcaacgacgt gagcccctcg 600 ggcggcttcg tggccgagct cctgcggcca gaattggacc cggtgtacat tccgccgcag 660 cagccgcagc cgccaggtgg cgggctgatg ggcaagttcg tgctgaaggc gtcgctgagc 720 gcccctggca gcgagtacgg cagcccgtcg gtcatcagcg tcagcaaagg cagccctgac 780 ggcagccacc cggtggtggt ggcgccctac aacggcgggc cgccgcgcac gtgccccaag 840 atcaagcagg aggcggtctc ttcgtgcacc cacttgggcg ctggaccccc tctcagcaat 900 ggccaccggc cggctgcaca cgacttcccc ctggggcggc agctccccag caggactacc 960 ccgaccctgg gtcttgagga agtgctgagc agcagggact gtcaccctgc cctgccgctt 1020 cctcccggct tccatcccca cccggggccc aattacccat ccttcctgcc cgatcagatg 1080 cagccgcaag tcccgccgct ccattaccaa gagctcatgc cacccggttc ctgcatgcca 1140 gaggagccca agccaaagag gggaagacga tcgtggcccc ggaaaaggac cgccacccac 1200 acttgtgatt acgcgggctg cggcaaaacc tacacaaaga gttcccatct caaggcacac 1260 ctgcgaaccc acacaggtga gaaaccttac cactgtgact gggacggctg tggatggaaa 1320 ttcgcccgct cagatgaact gaccaggcac taccgtaaac acacggggca ccgcccgttc 1380 cagtgccaaa aatgcgaccg agcattttcc aggtcggacc acctcgcctt acacatgaag 1440 aggcattttc tggcggtgct ggcggcggcg ccgtaa 1476 <210> 116 <211> 491 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Klf4-JO-86 MTD Amino Acid Sequence <400> 116 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Ala Val 1 5 10 15 Ser Asp Ala Leu Leu Pro Ser Phe Ser Thr Phe Ala Ser Gly Pro Ala 20 25 30 Gly Arg Glu Lys Thr Leu Arg Gln Ala Gly Ala Pro Asn Asn Arg Trp 35 40 45 Arg Glu Glu Leu Ser His Met Lys Arg Leu Pro Pro Val Leu Pro Gly 50 55 60 Arg Pro Tyr Asp Leu Ala Ala Ala Thr Val Ala Thr Asp Leu Glu Ser 65 70 75 80 Gly Gly Ala Gly Ala Ala Cys Gly Gly Ser Asn Leu Ala Pro Leu Pro 85 90 95 Arg Arg Glu Thr Glu Glu Phe Asn Asp Leu Leu Asp Leu Asp Phe Ile 100 105 110 Leu Ser Asn Ser Leu Thr His Pro Pro Glu Ser Val Ala Ala Thr Val 115 120 125 Ser Ser Ser Ala Ser Ala Ser Ser Ser Ser Ser Pro Ser Ser Ser Gly 130 135 140 Pro Ala Ser Ala Pro Ser Thr Cys Ser Phe Thr Tyr Pro Ile Arg Ala 145 150 155 160 Gly Asn Asp Pro Gly Val Ala Pro Gly Gly Thr Gly Gly Gly Leu Leu 165 170 175 Tyr Gly Arg Glu Ser Ala Pro Pro Pro Thr Ala Pro Phe Asn Leu Ala 180 185 190 Asp Ile Asn Asp Val Ser Pro Ser Gly Gly Phe Val Ala Glu Leu Leu 195 200 205 Arg Pro Glu Leu Asp Pro Val Tyr Ile Pro Pro Gln Gln Pro Gln Pro 210 215 220 Pro Gly Gly Gly Leu Met Gly Lys Phe Val Leu Lys Ala Ser Leu Ser 225 230 235 240 Ala Pro Gly Ser Glu Tyr Gly Ser Pro Ser Val Ile Ser Val Ser Lys 245 250 255 Gly Ser Pro Asp Gly Ser His Pro Val Val Val Ala Pro Tyr Asn Gly 260 265 270 Gly Pro Pro Arg Thr Cys Pro Lys Ile Lys Gln Glu Ala Val Ser Ser 275 280 285 Cys Thr His Leu Gly Ala Gly Pro Pro Leu Ser Asn Gly His Arg Pro 290 295 300 Ala Ala His Asp Phe Pro Leu Gly Arg Gln Leu Pro Ser Arg Thr Thr 305 310 315 320 Pro Thr Leu Gly Leu Glu Glu Val Leu Ser Ser Arg Asp Cys His Pro 325 330 335 Ala Leu Pro Leu Pro Pro Gly Phe His Pro His Pro Gly Pro Asn Tyr 340 345 350 Pro Ser Phe Leu Pro Asp Gln Met Gln Pro Gln Val Pro Pro Leu His 355 360 365 Tyr Gln Glu Leu Met Pro Pro Gly Ser Cys Met Pro Glu Glu Pro Lys 370 375 380 Pro Lys Arg Gly Arg Arg Ser Trp Pro Arg Lys Arg Thr Ala Thr His 385 390 395 400 Thr Cys Asp Tyr Ala Gly Cys Gly Lys Thr Tyr Thr Lys Ser Ser His 405 410 415 Leu Lys Ala His Leu Arg Thr His Thr Gly Glu Lys Pro Tyr His Cys 420 425 430 Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu Leu Thr 435 440 445 Arg His Tyr Arg Lys His Thr Gly His Arg Pro Phe Gln Cys Gln Lys 450 455 460 Cys Asp Arg Ala Phe Ser Arg Ser Asp His Leu Ala Leu His Met Lys 465 470 475 480 Arg His Phe Leu Ala Val Leu Ala Ala Ala Pro 485 490 <210> 117 <211> 1488 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Klf4 cDNA Sequence <400> 117 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaaggc tgtcagcgac gcgctgctcc catctttctc cacgttcgcg 120 tctggcccgg cgggaaggga gaagacactg cgtcaagcag gtgccccgaa taaccgctgg 180 cgggaggagc tctcccacat gaagcgactt cccccagtgc ttcccggccg cccctatgac 240 ctggcggcgg cgaccgtggc cacagacctg gagagcggcg gagccggtgc ggcttgcggc 300 ggtagcaacc tggcgcccct acctcggaga gagaccgagg agttcaacga tctcctggac 360 ctggacttta ttctctccaa ttcgctgacc catcctccgg agtcagtggc cgccaccgtg 420 tcctcgtcag cgtcagcctc ctcttcgtcg tcgccgtcga gcagcggccc tgccagcgcg 480 ccctccacct gcagcttcac ctatccgatc cgggccggga acgacccggg cgtggcgccg 540 ggcggcacgg gcggaggcct cctctatggc agggagtccg ctccccctcc gacggctccc 600 ttcaacctgg cggacatcaa cgacgtgagc ccctcgggcg gcttcgtggc cgagctcctg 660 cggccagaat tggacccggt gtacattccg ccgcagcagc cgcagccgcc aggtggcggg 720 ctgatgggca agttcgtgct gaaggcgtcg ctgagcgccc ctggcagcga gtacggcagc 780 ccgtcggtca tcagcgtcag caaaggcagc cctgacggca gccacccggt ggtggtggcg 840 ccctacaacg gcgggccgcc gcgcacgtgc cccaagatca agcaggaggc ggtctcttcg 900 tgcacccact tgggcgctgg accccctctc agcaatggcc accggccggc tgcacacgac 960 ttccccctgg ggcggcagct ccccagcagg actaccccga ccctgggtct tgaggaagtg 1020 ctgagcagca gggactgtca ccctgccctg ccgcttcctc ccggcttcca tccccacccg 1080 gggcccaatt acccatcctt cctgcccgat cagatgcagc cgcaagtccc gccgctccat 1140 taccaagagc tcatgccacc cggttcctgc atgccagagg agcccaagcc aaagagggga 1200 agacgatcgt ggccccggaa aaggaccgcc acccacactt gtgattacgc gggctgcggc 1260 aaaacctaca caaagagttc ccatctcaag gcacacctgc gaacccacac aggtgagaaa 1320 ccttaccact gtgactggga cggctgtgga tggaaattcg cccgctcaga tgaactgacc 1380 aggcactacc gtaaacacac ggggcaccgc ccgttccagt gccaaaaatg cgaccgagca 1440 ttttccaggt cggaccacct cgccttacac atgaagaggc atttttaa 1488 <210> 118 <211> 495 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Klf4 Amino Acid Sequence <400> 118 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Ala Val Ser Asp Ala Leu 20 25 30 Leu Pro Ser Phe Ser Thr Phe Ala Ser Gly Pro Ala Gly Arg Glu Lys 35 40 45 Thr Leu Arg Gln Ala Gly Ala Pro Asn Asn Arg Trp Arg Glu Glu Leu 50 55 60 Ser His Met Lys Arg Leu Pro Pro Val Leu Pro Gly Arg Pro Tyr Asp 65 70 75 80 Leu Ala Ala Ala Thr Val Ala Thr Asp Leu Glu Ser Gly Gly Ala Gly 85 90 95 Ala Ala Cys Gly Gly Ser Asn Leu Ala Pro Leu Pro Arg Arg Glu Thr 100 105 110 Glu Glu Phe Asn Asp Leu Leu Asp Leu Asp Phe Ile Leu Ser Asn Ser 115 120 125 Leu Thr His Pro Pro Glu Ser Val Ala Ala Thr Val Ser Ser Ser Ala 130 135 140 Ser Ala Ser Ser Ser Ser Ser Pro Ser Ser Ser Gly Pro Ala Ser Ala 145 150 155 160 Pro Ser Thr Cys Ser Phe Thr Tyr Pro Ile Arg Ala Gly Asn Asp Pro 165 170 175 Gly Val Ala Pro Gly Gly Thr Gly Gly Gly Leu Leu Tyr Gly Arg Glu 180 185 190 Ser Ala Pro Pro Pro Thr Ala Pro Phe Asn Leu Ala Asp Ile Asn Asp 195 200 205 Val Ser Pro Ser Gly Gly Phe Val Ala Glu Leu Leu Arg Pro Glu Leu 210 215 220 Asp Pro Val Tyr Ile Pro Pro Gln Gln Pro Gln Pro Pro Gly Gly Gly 225 230 235 240 Leu Met Gly Lys Phe Val Leu Lys Ala Ser Leu Ser Ala Pro Gly Ser 245 250 255 Glu Tyr Gly Ser Pro Ser Val Ile Ser Val Ser Lys Gly Ser Pro Asp 260 265 270 Gly Ser His Pro Val Val Val Ala Pro Tyr Asn Gly Gly Pro Pro Arg 275 280 285 Thr Cys Pro Lys Ile Lys Gln Glu Ala Val Ser Ser Cys Thr His Leu 290 295 300 Gly Ala Gly Pro Pro Leu Ser Asn Gly His Arg Pro Ala Ala His Asp 305 310 315 320 Phe Pro Leu Gly Arg Gln Leu Pro Ser Arg Thr Thr Pro Thr Leu Gly 325 330 335 Leu Glu Glu Val Leu Ser Ser Arg Asp Cys His Pro Ala Leu Pro Leu 340 345 350 Pro Pro Gly Phe His Pro His Pro Gly Pro Asn Tyr Pro Ser Phe Leu 355 360 365 Pro Asp Gln Met Gln Pro Gln Val Pro Pro Leu His Tyr Gln Glu Leu 370 375 380 Met Pro Pro Gly Ser Cys Met Pro Glu Glu Pro Lys Pro Lys Arg Gly 385 390 395 400 Arg Arg Ser Trp Pro Arg Lys Arg Thr Ala Thr His Thr Cys Asp Tyr 405 410 415 Ala Gly Cys Gly Lys Thr Tyr Thr Lys Ser Ser His Leu Lys Ala His 420 425 430 Leu Arg Thr His Thr Gly Glu Lys Pro Tyr His Cys Asp Trp Asp Gly 435 440 445 Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu Leu Thr Arg His Tyr Arg 450 455 460 Lys His Thr Gly His Arg Pro Phe Gln Cys Gln Lys Cys Asp Arg Ala 465 470 475 480 Phe Ser Arg Ser Asp His Leu Ala Leu His Met Lys Arg His Phe 485 490 495 <210> 119 <211> 1515 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Klf4 cDNA Sequence <400> 119 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctggctgt cagcgacgcg 120 ctgctcccat ctttctccac gttcgcgtct ggcccggcgg gaagggagaa gacactgcgt 180 caagcaggtg ccccgaataa ccgctggcgg gaggagctct cccacatgaa gcgacttccc 240 ccagtgcttc ccggccgccc ctatgacctg gcggcggcga ccgtggccac agacctggag 300 agcggcggag ccggtgcggc ttgcggcggt agcaacctgg cgcccctacc tcggagagag 360 accgaggagt tcaacgatct cctggacctg gactttattc tctccaattc gctgacccat 420 cctccggagt cagtggccgc caccgtgtcc tcgtcagcgt cagcctcctc ttcgtcgtcg 480 ccgtcgagca gcggccctgc cagcgcgccc tccacctgca gcttcaccta tccgatccgg 540 gccgggaacg acccgggcgt ggcgccgggc ggcacgggcg gaggcctcct ctatggcagg 600 gagtccgctc cccctccgac ggctcccttc aacctggcgg acatcaacga cgtgagcccc 660 tcgggcggct tcgtggccga gctcctgcgg ccagaattgg acccggtgta cattccgccg 720 cagcagccgc agccgccagg tggcgggctg atgggcaagt tcgtgctgaa ggcgtcgctg 780 agcgcccctg gcagcgagta cggcagcccg tcggtcatca gcgtcagcaa aggcagccct 840 gacggcagcc acccggtggt ggtggcgccc tacaacggcg ggccgccgcg cacgtgcccc 900 aagatcaagc aggaggcggt ctcttcgtgc acccacttgg gcgctggacc ccctctcagc 960 aatggccacc ggccggctgc acacgacttc cccctggggc ggcagctccc cagcaggact 1020 accccgaccc tgggtcttga ggaagtgctg agcagcaggg actgtcaccc tgccctgccg 1080 cttcctcccg gcttccatcc ccacccgggg cccaattacc catccttcct gcccgatcag 1140 atgcagccgc aagtcccgcc gctccattac caagagctca tgccacccgg ttcctgcatg 1200 ccagaggagc ccaagccaaa gaggggaaga cgatcgtggc cccggaaaag gaccgccacc 1260 cacacttgtg attacgcggg ctgcggcaaa acctacacaa agagttccca tctcaaggca 1320 cacctgcgaa cccacacagg tgagaaacct taccactgtg actgggacgg ctgtggatgg 1380 aaattcgccc gctcagatga actgaccagg cactaccgta aacacacggg gcaccgcccg 1440 ttccagtgcc aaaaatgcga ccgagcattt tccaggtcgg accacctcgc cttacacatg 1500 aagaggcatt tttaa 1515 <210> 120 <211> 504 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Klf4 Amino Acid Sequence <400> 120 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Ala Val Ser Asp Ala Leu Leu Pro Ser Phe Ser Thr Phe 35 40 45 Ala Ser Gly Pro Ala Gly Arg Glu Lys Thr Leu Arg Gln Ala Gly Ala 50 55 60 Pro Asn Asn Arg Trp Arg Glu Glu Leu Ser His Met Lys Arg Leu Pro 65 70 75 80 Pro Val Leu Pro Gly Arg Pro Tyr Asp Leu Ala Ala Ala Thr Val Ala 85 90 95 Thr Asp Leu Glu Ser Gly Gly Ala Gly Ala Ala Cys Gly Gly Ser Asn 100 105 110 Leu Ala Pro Leu Pro Arg Arg Glu Thr Glu Glu Phe Asn Asp Leu Leu 115 120 125 Asp Leu Asp Phe Ile Leu Ser Asn Ser Leu Thr His Pro Pro Glu Ser 130 135 140 Val Ala Ala Thr Val Ser Ser Ser Ala Ser Ala Ser Ser Ser Ser Ser 145 150 155 160 Pro Ser Ser Ser Gly Pro Ala Ser Ala Pro Ser Thr Cys Ser Phe Thr 165 170 175 Tyr Pro Ile Arg Ala Gly Asn Asp Pro Gly Val Ala Pro Gly Gly Thr 180 185 190 Gly Gly Gly Leu Leu Tyr Gly Arg Glu Ser Ala Pro Pro Pro Thr Ala 195 200 205 Pro Phe Asn Leu Ala Asp Ile Asn Asp Val Ser Pro Ser Gly Gly Phe 210 215 220 Val Ala Glu Leu Leu Arg Pro Glu Leu Asp Pro Val Tyr Ile Pro Pro 225 230 235 240 Gln Gln Pro Gln Pro Pro Gly Gly Gly Leu Met Gly Lys Phe Val Leu 245 250 255 Lys Ala Ser Leu Ser Ala Pro Gly Ser Glu Tyr Gly Ser Pro Ser Val 260 265 270 Ile Ser Val Ser Lys Gly Ser Pro Asp Gly Ser His Pro Val Val Val 275 280 285 Ala Pro Tyr Asn Gly Gly Pro Pro Arg Thr Cys Pro Lys Ile Lys Gln 290 295 300 Glu Ala Val Ser Ser Cys Thr His Leu Gly Ala Gly Pro Pro Leu Ser 305 310 315 320 Asn Gly His Arg Pro Ala Ala His Asp Phe Pro Leu Gly Arg Gln Leu 325 330 335 Pro Ser Arg Thr Thr Pro Thr Leu Gly Leu Glu Glu Val Leu Ser Ser 340 345 350 Arg Asp Cys His Pro Ala Leu Pro Leu Pro Pro Gly Phe His Pro His 355 360 365 Pro Gly Pro Asn Tyr Pro Ser Phe Leu Pro Asp Gln Met Gln Pro Gln 370 375 380 Val Pro Pro Leu His Tyr Gln Glu Leu Met Pro Pro Gly Ser Cys Met 385 390 395 400 Pro Glu Glu Pro Lys Pro Lys Arg Gly Arg Arg Ser Trp Pro Arg Lys 405 410 415 Arg Thr Ala Thr His Thr Cys Asp Tyr Ala Gly Cys Gly Lys Thr Tyr 420 425 430 Thr Lys Ser Ser His Leu Lys Ala His Leu Arg Thr His Thr Gly Glu 435 440 445 Lys Pro Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg 450 455 460 Ser Asp Glu Leu Thr Arg His Tyr Arg Lys His Thr Gly His Arg Pro 465 470 475 480 Phe Gln Cys Gln Lys Cys Asp Arg Ala Phe Ser Arg Ser Asp His Leu 485 490 495 Ala Leu His Met Lys Arg His Phe 500 <210> 121 <211> 1515 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Klf4-JO-84 MTD cDNA Sequence <400> 121 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaaggc tgtcagcgac gcgctgctcc catctttctc cacgttcgcg 120 tctggcccgg cgggaaggga gaagacactg cgtcaagcag gtgccccgaa taaccgctgg 180 cgggaggagc tctcccacat gaagcgactt cccccagtgc ttcccggccg cccctatgac 240 ctggcggcgg cgaccgtggc cacagacctg gagagcggcg gagccggtgc ggcttgcggc 300 ggtagcaacc tggcgcccct acctcggaga gagaccgagg agttcaacga tctcctggac 360 ctggacttta ttctctccaa ttcgctgacc catcctccgg agtcagtggc cgccaccgtg 420 tcctcgtcag cgtcagcctc ctcttcgtcg tcgccgtcga gcagcggccc tgccagcgcg 480 ccctccacct gcagcttcac ctatccgatc cgggccggga acgacccggg cgtggcgccg 540 ggcggcacgg gcggaggcct cctctatggc agggagtccg ctccccctcc gacggctccc 600 ttcaacctgg cggacatcaa cgacgtgagc ccctcgggcg gcttcgtggc cgagctcctg 660 cggccagaat tggacccggt gtacattccg ccgcagcagc cgcagccgcc aggtggcggg 720 ctgatgggca agttcgtgct gaaggcgtcg ctgagcgccc ctggcagcga gtacggcagc 780 ccgtcggtca tcagcgtcag caaaggcagc cctgacggca gccacccggt ggtggtggcg 840 ccctacaacg gcgggccgcc gcgcacgtgc cccaagatca agcaggaggc ggtctcttcg 900 tgcacccact tgggcgctgg accccctctc agcaatggcc accggccggc tgcacacgac 960 ttccccctgg ggcggcagct ccccagcagg actaccccga ccctgggtct tgaggaagtg 1020 ctgagcagca gggactgtca ccctgccctg ccgcttcctc ccggcttcca tccccacccg 1080 gggcccaatt acccatcctt cctgcccgat cagatgcagc cgcaagtccc gccgctccat 1140 taccaagagc tcatgccacc cggttcctgc atgccagagg agcccaagcc aaagagggga 1200 agacgatcgt ggccccggaa aaggaccgcc acccacactt gtgattacgc gggctgcggc 1260 aaaacctaca caaagagttc ccatctcaag gcacacctgc gaacccacac aggtgagaaa 1320 ccttaccact gtgactggga cggctgtgga tggaaattcg cccgctcaga tgaactgacc 1380 aggcactacc gtaaacacac ggggcaccgc ccgttccagt gccaaaaatg cgaccgagca 1440 ttttccaggt cggaccacct cgccttacac atgaagaggc attttctggt ggcggcgctg 1500 ctggcggtgc tgtaa 1515 <210> 122 <211> 504 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Klf4-JO-84 MTD Amino Acid Sequence <400> 122 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Ala Val Ser Asp Ala Leu 20 25 30 Leu Pro Ser Phe Ser Thr Phe Ala Ser Gly Pro Ala Gly Arg Glu Lys 35 40 45 Thr Leu Arg Gln Ala Gly Ala Pro Asn Asn Arg Trp Arg Glu Glu Leu 50 55 60 Ser His Met Lys Arg Leu Pro Pro Val Leu Pro Gly Arg Pro Tyr Asp 65 70 75 80 Leu Ala Ala Ala Thr Val Ala Thr Asp Leu Glu Ser Gly Gly Ala Gly 85 90 95 Ala Ala Cys Gly Gly Ser Asn Leu Ala Pro Leu Pro Arg Arg Glu Thr 100 105 110 Glu Glu Phe Asn Asp Leu Leu Asp Leu Asp Phe Ile Leu Ser Asn Ser 115 120 125 Leu Thr His Pro Pro Glu Ser Val Ala Ala Thr Val Ser Ser Ser Ala 130 135 140 Ser Ala Ser Ser Ser Ser Ser Pro Ser Ser Ser Gly Pro Ala Ser Ala 145 150 155 160 Pro Ser Thr Cys Ser Phe Thr Tyr Pro Ile Arg Ala Gly Asn Asp Pro 165 170 175 Gly Val Ala Pro Gly Gly Thr Gly Gly Gly Leu Leu Tyr Gly Arg Glu 180 185 190 Ser Ala Pro Pro Pro Thr Ala Pro Phe Asn Leu Ala Asp Ile Asn Asp 195 200 205 Val Ser Pro Ser Gly Gly Phe Val Ala Glu Leu Leu Arg Pro Glu Leu 210 215 220 Asp Pro Val Tyr Ile Pro Pro Gln Gln Pro Gln Pro Pro Gly Gly Gly 225 230 235 240 Leu Met Gly Lys Phe Val Leu Lys Ala Ser Leu Ser Ala Pro Gly Ser 245 250 255 Glu Tyr Gly Ser Pro Ser Val Ile Ser Val Ser Lys Gly Ser Pro Asp 260 265 270 Gly Ser His Pro Val Val Val Ala Pro Tyr Asn Gly Gly Pro Pro Arg 275 280 285 Thr Cys Pro Lys Ile Lys Gln Glu Ala Val Ser Ser Cys Thr His Leu 290 295 300 Gly Ala Gly Pro Pro Leu Ser Asn Gly His Arg Pro Ala Ala His Asp 305 310 315 320 Phe Pro Leu Gly Arg Gln Leu Pro Ser Arg Thr Thr Pro Thr Leu Gly 325 330 335 Leu Glu Glu Val Leu Ser Ser Arg Asp Cys His Pro Ala Leu Pro Leu 340 345 350 Pro Pro Gly Phe His Pro His Pro Gly Pro Asn Tyr Pro Ser Phe Leu 355 360 365 Pro Asp Gln Met Gln Pro Gln Val Pro Pro Leu His Tyr Gln Glu Leu 370 375 380 Met Pro Pro Gly Ser Cys Met Pro Glu Glu Pro Lys Pro Lys Arg Gly 385 390 395 400 Arg Arg Ser Trp Pro Arg Lys Arg Thr Ala Thr His Thr Cys Asp Tyr 405 410 415 Ala Gly Cys Gly Lys Thr Tyr Thr Lys Ser Ser His Leu Lys Ala His 420 425 430 Leu Arg Thr His Thr Gly Glu Lys Pro Tyr His Cys Asp Trp Asp Gly 435 440 445 Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu Leu Thr Arg His Tyr Arg 450 455 460 Lys His Thr Gly His Arg Pro Phe Gln Cys Gln Lys Cys Asp Arg Ala 465 470 475 480 Phe Ser Arg Ser Asp His Leu Ala Leu His Met Lys Arg His Phe Leu 485 490 495 Val Ala Ala Leu Leu Ala Val Leu 500 <210> 123 <211> 1542 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Klf4-JO-84 MTD cDNA Sequence <400> 123 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctggctgt cagcgacgcg 120 ctgctcccat ctttctccac gttcgcgtct ggcccggcgg gaagggagaa gacactgcgt 180 caagcaggtg ccccgaataa ccgctggcgg gaggagctct cccacatgaa gcgacttccc 240 ccagtgcttc ccggccgccc ctatgacctg gcggcggcga ccgtggccac agacctggag 300 agcggcggag ccggtgcggc ttgcggcggt agcaacctgg cgcccctacc tcggagagag 360 accgaggagt tcaacgatct cctggacctg gactttattc tctccaattc gctgacccat 420 cctccggagt cagtggccgc caccgtgtcc tcgtcagcgt cagcctcctc ttcgtcgtcg 480 ccgtcgagca gcggccctgc cagcgcgccc tccacctgca gcttcaccta tccgatccgg 540 gccgggaacg acccgggcgt ggcgccgggc ggcacgggcg gaggcctcct ctatggcagg 600 gagtccgctc cccctccgac ggctcccttc aacctggcgg acatcaacga cgtgagcccc 660 tcgggcggct tcgtggccga gctcctgcgg ccagaattgg acccggtgta cattccgccg 720 cagcagccgc agccgccagg tggcgggctg atgggcaagt tcgtgctgaa ggcgtcgctg 780 agcgcccctg gcagcgagta cggcagcccg tcggtcatca gcgtcagcaa aggcagccct 840 gacggcagcc acccggtggt ggtggcgccc tacaacggcg ggccgccgcg cacgtgcccc 900 aagatcaagc aggaggcggt ctcttcgtgc acccacttgg gcgctggacc ccctctcagc 960 aatggccacc ggccggctgc acacgacttc cccctggggc ggcagctccc cagcaggact 1020 accccgaccc tgggtcttga ggaagtgctg agcagcaggg actgtcaccc tgccctgccg 1080 cttcctcccg gcttccatcc ccacccgggg cccaattacc catccttcct gcccgatcag 1140 atgcagccgc aagtcccgcc gctccattac caagagctca tgccacccgg ttcctgcatg 1200 ccagaggagc ccaagccaaa gaggggaaga cgatcgtggc cccggaaaag gaccgccacc 1260 cacacttgtg attacgcggg ctgcggcaaa acctacacaa agagttccca tctcaaggca 1320 cacctgcgaa cccacacagg tgagaaacct taccactgtg actgggacgg ctgtggatgg 1380 aaattcgccc gctcagatga actgaccagg cactaccgta aacacacggg gcaccgcccg 1440 ttccagtgcc aaaaatgcga ccgagcattt tccaggtcgg accacctcgc cttacacatg 1500 aagaggcatt ttctggtggc ggcgctgctg gcggtgctgt aa 1542 <210> 124 <211> 513 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Klf4-JO-84 MTD Amino Acid Sequence <400> 124 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Ala Val Ser Asp Ala Leu Leu Pro Ser Phe Ser Thr Phe 35 40 45 Ala Ser Gly Pro Ala Gly Arg Glu Lys Thr Leu Arg Gln Ala Gly Ala 50 55 60 Pro Asn Asn Arg Trp Arg Glu Glu Leu Ser His Met Lys Arg Leu Pro 65 70 75 80 Pro Val Leu Pro Gly Arg Pro Tyr Asp Leu Ala Ala Ala Thr Val Ala 85 90 95 Thr Asp Leu Glu Ser Gly Gly Ala Gly Ala Ala Cys Gly Gly Ser Asn 100 105 110 Leu Ala Pro Leu Pro Arg Arg Glu Thr Glu Glu Phe Asn Asp Leu Leu 115 120 125 Asp Leu Asp Phe Ile Leu Ser Asn Ser Leu Thr His Pro Pro Glu Ser 130 135 140 Val Ala Ala Thr Val Ser Ser Ser Ala Ser Ala Ser Ser Ser Ser Ser 145 150 155 160 Pro Ser Ser Ser Gly Pro Ala Ser Ala Pro Ser Thr Cys Ser Phe Thr 165 170 175 Tyr Pro Ile Arg Ala Gly Asn Asp Pro Gly Val Ala Pro Gly Gly Thr 180 185 190 Gly Gly Gly Leu Leu Tyr Gly Arg Glu Ser Ala Pro Pro Pro Thr Ala 195 200 205 Pro Phe Asn Leu Ala Asp Ile Asn Asp Val Ser Pro Ser Gly Gly Phe 210 215 220 Val Ala Glu Leu Leu Arg Pro Glu Leu Asp Pro Val Tyr Ile Pro Pro 225 230 235 240 Gln Gln Pro Gln Pro Pro Gly Gly Gly Leu Met Gly Lys Phe Val Leu 245 250 255 Lys Ala Ser Leu Ser Ala Pro Gly Ser Glu Tyr Gly Ser Pro Ser Val 260 265 270 Ile Ser Val Ser Lys Gly Ser Pro Asp Gly Ser His Pro Val Val Val 275 280 285 Ala Pro Tyr Asn Gly Gly Pro Pro Arg Thr Cys Pro Lys Ile Lys Gln 290 295 300 Glu Ala Val Ser Ser Cys Thr His Leu Gly Ala Gly Pro Pro Leu Ser 305 310 315 320 Asn Gly His Arg Pro Ala Ala His Asp Phe Pro Leu Gly Arg Gln Leu 325 330 335 Pro Ser Arg Thr Thr Pro Thr Leu Gly Leu Glu Glu Val Leu Ser Ser 340 345 350 Arg Asp Cys His Pro Ala Leu Pro Leu Pro Pro Gly Phe His Pro His 355 360 365 Pro Gly Pro Asn Tyr Pro Ser Phe Leu Pro Asp Gln Met Gln Pro Gln 370 375 380 Val Pro Pro Leu His Tyr Gln Glu Leu Met Pro Pro Gly Ser Cys Met 385 390 395 400 Pro Glu Glu Pro Lys Pro Lys Arg Gly Arg Arg Ser Trp Pro Arg Lys 405 410 415 Arg Thr Ala Thr His Thr Cys Asp Tyr Ala Gly Cys Gly Lys Thr Tyr 420 425 430 Thr Lys Ser Ser His Leu Lys Ala His Leu Arg Thr His Thr Gly Glu 435 440 445 Lys Pro Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg 450 455 460 Ser Asp Glu Leu Thr Arg His Tyr Arg Lys His Thr Gly His Arg Pro 465 470 475 480 Phe Gln Cys Gln Lys Cys Asp Arg Ala Phe Ser Arg Ser Asp His Leu 485 490 495 Ala Leu His Met Lys Arg His Phe Leu Val Ala Ala Leu Leu Ala Val 500 505 510 Leu <210> 125 <211> 1512 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Klf4 cDNA Sequence <400> 125 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cggctgtcag cgacgcgctg 120 ctcccatctt tctccacgtt cgcgtctggc ccggcgggaa gggagaagac actgcgtcaa 180 gcaggtgccc cgaataaccg ctggcgggag gagctctccc acatgaagcg acttccccca 240 gtgcttcccg gccgccccta tgacctggcg gcggcgaccg tggccacaga cctggagagc 300 ggcggagccg gtgcggcttg cggcggtagc aacctggcgc ccctacctcg gagagagacc 360 gaggagttca acgatctcct ggacctggac tttattctct ccaattcgct gacccatcct 420 ccggagtcag tggccgccac cgtgtcctcg tcagcgtcag cctcctcttc gtcgtcgccg 480 tcgagcagcg gccctgccag cgcgccctcc acctgcagct tcacctatcc gatccgggcc 540 gggaacgacc cgggcgtggc gccgggcggc acgggcggag gcctcctcta tggcagggag 600 tccgctcccc ctccgacggc tcccttcaac ctggcggaca tcaacgacgt gagcccctcg 660 ggcggcttcg tggccgagct cctgcggcca gaattggacc cggtgtacat tccgccgcag 720 cagccgcagc cgccaggtgg cgggctgatg ggcaagttcg tgctgaaggc gtcgctgagc 780 gcccctggca gcgagtacgg cagcccgtcg gtcatcagcg tcagcaaagg cagccctgac 840 ggcagccacc cggtggtggt ggcgccctac aacggcgggc cgccgcgcac gtgccccaag 900 atcaagcagg aggcggtctc ttcgtgcacc cacttgggcg ctggaccccc tctcagcaat 960 ggccaccggc cggctgcaca cgacttcccc ctggggcggc agctccccag caggactacc 1020 ccgaccctgg gtcttgagga agtgctgagc agcagggact gtcaccctgc cctgccgctt 1080 cctcccggct tccatcccca cccggggccc aattacccat ccttcctgcc cgatcagatg 1140 cagccgcaag tcccgccgct ccattaccaa gagctcatgc cacccggttc ctgcatgcca 1200 gaggagccca agccaaagag gggaagacga tcgtggcccc ggaaaaggac cgccacccac 1260 acttgtgatt acgcgggctg cggcaaaacc tacacaaaga gttcccatct caaggcacac 1320 ctgcgaaccc acacaggtga gaaaccttac cactgtgact gggacggctg tggatggaaa 1380 ttcgcccgct cagatgaact gaccaggcac taccgtaaac acacggggca ccgcccgttc 1440 cagtgccaaa aatgcgaccg agcattttcc aggtcggacc acctcgcctt acacatgaag 1500 aggcattttt aa 1512 <210> 126 <211> 503 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Klf4 Amino Acid Sequence <400> 126 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Ala Val Ser Asp Ala Leu Leu Pro Ser Phe Ser Thr Phe Ala 35 40 45 Ser Gly Pro Ala Gly Arg Glu Lys Thr Leu Arg Gln Ala Gly Ala Pro 50 55 60 Asn Asn Arg Trp Arg Glu Glu Leu Ser His Met Lys Arg Leu Pro Pro 65 70 75 80 Val Leu Pro Gly Arg Pro Tyr Asp Leu Ala Ala Ala Thr Val Ala Thr 85 90 95 Asp Leu Glu Ser Gly Gly Ala Gly Ala Ala Cys Gly Gly Ser Asn Leu 100 105 110 Ala Pro Leu Pro Arg Arg Glu Thr Glu Glu Phe Asn Asp Leu Leu Asp 115 120 125 Leu Asp Phe Ile Leu Ser Asn Ser Leu Thr His Pro Pro Glu Ser Val 130 135 140 Ala Ala Thr Val Ser Ser Ser Ala Ser Ala Ser Ser Ser Ser Ser Pro 145 150 155 160 Ser Ser Ser Gly Pro Ala Ser Ala Pro Ser Thr Cys Ser Phe Thr Tyr 165 170 175 Pro Ile Arg Ala Gly Asn Asp Pro Gly Val Ala Pro Gly Gly Thr Gly 180 185 190 Gly Gly Leu Leu Tyr Gly Arg Glu Ser Ala Pro Pro Pro Thr Ala Pro 195 200 205 Phe Asn Leu Ala Asp Ile Asn Asp Val Ser Pro Ser Gly Gly Phe Val 210 215 220 Ala Glu Leu Leu Arg Pro Glu Leu Asp Pro Val Tyr Ile Pro Pro Gln 225 230 235 240 Gln Pro Gln Pro Pro Gly Gly Gly Leu Met Gly Lys Phe Val Leu Lys 245 250 255 Ala Ser Leu Ser Ala Pro Gly Ser Glu Tyr Gly Ser Pro Ser Val Ile 260 265 270 Ser Val Ser Lys Gly Ser Pro Asp Gly Ser His Pro Val Val Val Ala 275 280 285 Pro Tyr Asn Gly Gly Pro Pro Arg Thr Cys Pro Lys Ile Lys Gln Glu 290 295 300 Ala Val Ser Ser Cys Thr His Leu Gly Ala Gly Pro Pro Leu Ser Asn 305 310 315 320 Gly His Arg Pro Ala Ala His Asp Phe Pro Leu Gly Arg Gln Leu Pro 325 330 335 Ser Arg Thr Thr Pro Thr Leu Gly Leu Glu Glu Val Leu Ser Ser Arg 340 345 350 Asp Cys His Pro Ala Leu Pro Leu Pro Pro Gly Phe His Pro His Pro 355 360 365 Gly Pro Asn Tyr Pro Ser Phe Leu Pro Asp Gln Met Gln Pro Gln Val 370 375 380 Pro Pro Leu His Tyr Gln Glu Leu Met Pro Pro Gly Ser Cys Met Pro 385 390 395 400 Glu Glu Pro Lys Pro Lys Arg Gly Arg Arg Ser Trp Pro Arg Lys Arg 405 410 415 Thr Ala Thr His Thr Cys Asp Tyr Ala Gly Cys Gly Lys Thr Tyr Thr 420 425 430 Lys Ser Ser His Leu Lys Ala His Leu Arg Thr His Thr Gly Glu Lys 435 440 445 Pro Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser 450 455 460 Asp Glu Leu Thr Arg His Tyr Arg Lys His Thr Gly His Arg Pro Phe 465 470 475 480 Gln Cys Gln Lys Cys Asp Arg Ala Phe Ser Arg Ser Asp His Leu Ala 485 490 495 Leu His Met Lys Arg His Phe 500 <210> 127 <211> 1512 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Klf4-JO-86 MTD cDNA Sequence <400> 127 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaaggc tgtcagcgac gcgctgctcc catctttctc cacgttcgcg 120 tctggcccgg cgggaaggga gaagacactg cgtcaagcag gtgccccgaa taaccgctgg 180 cgggaggagc tctcccacat gaagcgactt cccccagtgc ttcccggccg cccctatgac 240 ctggcggcgg cgaccgtggc cacagacctg gagagcggcg gagccggtgc ggcttgcggc 300 ggtagcaacc tggcgcccct acctcggaga gagaccgagg agttcaacga tctcctggac 360 ctggacttta ttctctccaa ttcgctgacc catcctccgg agtcagtggc cgccaccgtg 420 tcctcgtcag cgtcagcctc ctcttcgtcg tcgccgtcga gcagcggccc tgccagcgcg 480 ccctccacct gcagcttcac ctatccgatc cgggccggga acgacccggg cgtggcgccg 540 ggcggcacgg gcggaggcct cctctatggc agggagtccg ctccccctcc gacggctccc 600 ttcaacctgg cggacatcaa cgacgtgagc ccctcgggcg gcttcgtggc cgagctcctg 660 cggccagaat tggacccggt gtacattccg ccgcagcagc cgcagccgcc aggtggcggg 720 ctgatgggca agttcgtgct gaaggcgtcg ctgagcgccc ctggcagcga gtacggcagc 780 ccgtcggtca tcagcgtcag caaaggcagc cctgacggca gccacccggt ggtggtggcg 840 ccctacaacg gcgggccgcc gcgcacgtgc cccaagatca agcaggaggc ggtctcttcg 900 tgcacccact tgggcgctgg accccctctc agcaatggcc accggccggc tgcacacgac 960 ttccccctgg ggcggcagct ccccagcagg actaccccga ccctgggtct tgaggaagtg 1020 ctgagcagca gggactgtca ccctgccctg ccgcttcctc ccggcttcca tccccacccg 1080 gggcccaatt acccatcctt cctgcccgat cagatgcagc cgcaagtccc gccgctccat 1140 taccaagagc tcatgccacc cggttcctgc atgccagagg agcccaagcc aaagagggga 1200 agacgatcgt ggccccggaa aaggaccgcc acccacactt gtgattacgc gggctgcggc 1260 aaaacctaca caaagagttc ccatctcaag gcacacctgc gaacccacac aggtgagaaa 1320 ccttaccact gtgactggga cggctgtgga tggaaattcg cccgctcaga tgaactgacc 1380 aggcactacc gtaaacacac ggggcaccgc ccgttccagt gccaaaaatg cgaccgagca 1440 ttttccaggt cggaccacct cgccttacac atgaagaggc attttctggc ggtgctggcg 1500 gcggcgccgt aa 1512 <210> 128 <211> 503 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Klf4-JO-86 MTD Amino Acid Sequence <400> 128 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Ala Val Ser Asp Ala Leu 20 25 30 Leu Pro Ser Phe Ser Thr Phe Ala Ser Gly Pro Ala Gly Arg Glu Lys 35 40 45 Thr Leu Arg Gln Ala Gly Ala Pro Asn Asn Arg Trp Arg Glu Glu Leu 50 55 60 Ser His Met Lys Arg Leu Pro Pro Val Leu Pro Gly Arg Pro Tyr Asp 65 70 75 80 Leu Ala Ala Ala Thr Val Ala Thr Asp Leu Glu Ser Gly Gly Ala Gly 85 90 95 Ala Ala Cys Gly Gly Ser Asn Leu Ala Pro Leu Pro Arg Arg Glu Thr 100 105 110 Glu Glu Phe Asn Asp Leu Leu Asp Leu Asp Phe Ile Leu Ser Asn Ser 115 120 125 Leu Thr His Pro Pro Glu Ser Val Ala Ala Thr Val Ser Ser Ser Ala 130 135 140 Ser Ala Ser Ser Ser Ser Ser Pro Ser Ser Ser Gly Pro Ala Ser Ala 145 150 155 160 Pro Ser Thr Cys Ser Phe Thr Tyr Pro Ile Arg Ala Gly Asn Asp Pro 165 170 175 Gly Val Ala Pro Gly Gly Thr Gly Gly Gly Leu Leu Tyr Gly Arg Glu 180 185 190 Ser Ala Pro Pro Pro Thr Ala Pro Phe Asn Leu Ala Asp Ile Asn Asp 195 200 205 Val Ser Pro Ser Gly Gly Phe Val Ala Glu Leu Leu Arg Pro Glu Leu 210 215 220 Asp Pro Val Tyr Ile Pro Pro Gln Gln Pro Gln Pro Pro Gly Gly Gly 225 230 235 240 Leu Met Gly Lys Phe Val Leu Lys Ala Ser Leu Ser Ala Pro Gly Ser 245 250 255 Glu Tyr Gly Ser Pro Ser Val Ile Ser Val Ser Lys Gly Ser Pro Asp 260 265 270 Gly Ser His Pro Val Val Val Ala Pro Tyr Asn Gly Gly Pro Pro Arg 275 280 285 Thr Cys Pro Lys Ile Lys Gln Glu Ala Val Ser Ser Cys Thr His Leu 290 295 300 Gly Ala Gly Pro Pro Leu Ser Asn Gly His Arg Pro Ala Ala His Asp 305 310 315 320 Phe Pro Leu Gly Arg Gln Leu Pro Ser Arg Thr Thr Pro Thr Leu Gly 325 330 335 Leu Glu Glu Val Leu Ser Ser Arg Asp Cys His Pro Ala Leu Pro Leu 340 345 350 Pro Pro Gly Phe His Pro His Pro Gly Pro Asn Tyr Pro Ser Phe Leu 355 360 365 Pro Asp Gln Met Gln Pro Gln Val Pro Pro Leu His Tyr Gln Glu Leu 370 375 380 Met Pro Pro Gly Ser Cys Met Pro Glu Glu Pro Lys Pro Lys Arg Gly 385 390 395 400 Arg Arg Ser Trp Pro Arg Lys Arg Thr Ala Thr His Thr Cys Asp Tyr 405 410 415 Ala Gly Cys Gly Lys Thr Tyr Thr Lys Ser Ser His Leu Lys Ala His 420 425 430 Leu Arg Thr His Thr Gly Glu Lys Pro Tyr His Cys Asp Trp Asp Gly 435 440 445 Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu Leu Thr Arg His Tyr Arg 450 455 460 Lys His Thr Gly His Arg Pro Phe Gln Cys Gln Lys Cys Asp Arg Ala 465 470 475 480 Phe Ser Arg Ser Asp His Leu Ala Leu His Met Lys Arg His Phe Leu 485 490 495 Ala Val Leu Ala Ala Ala Pro 500 <210> 129 <211> 1536 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Klf4-JO-86 MTD cDNA Sequence <400> 129 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cggctgtcag cgacgcgctg 120 ctcccatctt tctccacgtt cgcgtctggc ccggcgggaa gggagaagac actgcgtcaa 180 gcaggtgccc cgaataaccg ctggcgggag gagctctccc acatgaagcg acttccccca 240 gtgcttcccg gccgccccta tgacctggcg gcggcgaccg tggccacaga cctggagagc 300 ggcggagccg gtgcggcttg cggcggtagc aacctggcgc ccctacctcg gagagagacc 360 gaggagttca acgatctcct ggacctggac tttattctct ccaattcgct gacccatcct 420 ccggagtcag tggccgccac cgtgtcctcg tcagcgtcag cctcctcttc gtcgtcgccg 480 tcgagcagcg gccctgccag cgcgccctcc acctgcagct tcacctatcc gatccgggcc 540 gggaacgacc cgggcgtggc gccgggcggc acgggcggag gcctcctcta tggcagggag 600 tccgctcccc ctccgacggc tcccttcaac ctggcggaca tcaacgacgt gagcccctcg 660 ggcggcttcg tggccgagct cctgcggcca gaattggacc cggtgtacat tccgccgcag 720 cagccgcagc cgccaggtgg cgggctgatg ggcaagttcg tgctgaaggc gtcgctgagc 780 gcccctggca gcgagtacgg cagcccgtcg gtcatcagcg tcagcaaagg cagccctgac 840 ggcagccacc cggtggtggt ggcgccctac aacggcgggc cgccgcgcac gtgccccaag 900 atcaagcagg aggcggtctc ttcgtgcacc cacttgggcg ctggaccccc tctcagcaat 960 ggccaccggc cggctgcaca cgacttcccc ctggggcggc agctccccag caggactacc 1020 ccgaccctgg gtcttgagga agtgctgagc agcagggact gtcaccctgc cctgccgctt 1080 cctcccggct tccatcccca cccggggccc aattacccat ccttcctgcc cgatcagatg 1140 cagccgcaag tcccgccgct ccattaccaa gagctcatgc cacccggttc ctgcatgcca 1200 gaggagccca agccaaagag gggaagacga tcgtggcccc ggaaaaggac cgccacccac 1260 acttgtgatt acgcgggctg cggcaaaacc tacacaaaga gttcccatct caaggcacac 1320 ctgcgaaccc acacaggtga gaaaccttac cactgtgact gggacggctg tggatggaaa 1380 ttcgcccgct cagatgaact gaccaggcac taccgtaaac acacggggca ccgcccgttc 1440 cagtgccaaa aatgcgaccg agcattttcc aggtcggacc acctcgcctt acacatgaag 1500 aggcattttc tggcggtgct ggcggcggcg ccgtaa 1536 <210> 130 <211> 511 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Klf4-JO-86 MTD Amino Acid Sequence <400> 130 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Ala Val Ser Asp Ala Leu Leu Pro Ser Phe Ser Thr Phe Ala 35 40 45 Ser Gly Pro Ala Gly Arg Glu Lys Thr Leu Arg Gln Ala Gly Ala Pro 50 55 60 Asn Asn Arg Trp Arg Glu Glu Leu Ser His Met Lys Arg Leu Pro Pro 65 70 75 80 Val Leu Pro Gly Arg Pro Tyr Asp Leu Ala Ala Ala Thr Val Ala Thr 85 90 95 Asp Leu Glu Ser Gly Gly Ala Gly Ala Ala Cys Gly Gly Ser Asn Leu 100 105 110 Ala Pro Leu Pro Arg Arg Glu Thr Glu Glu Phe Asn Asp Leu Leu Asp 115 120 125 Leu Asp Phe Ile Leu Ser Asn Ser Leu Thr His Pro Pro Glu Ser Val 130 135 140 Ala Ala Thr Val Ser Ser Ser Ala Ser Ala Ser Ser Ser Ser Ser Pro 145 150 155 160 Ser Ser Ser Gly Pro Ala Ser Ala Pro Ser Thr Cys Ser Phe Thr Tyr 165 170 175 Pro Ile Arg Ala Gly Asn Asp Pro Gly Val Ala Pro Gly Gly Thr Gly 180 185 190 Gly Gly Leu Leu Tyr Gly Arg Glu Ser Ala Pro Pro Pro Thr Ala Pro 195 200 205 Phe Asn Leu Ala Asp Ile Asn Asp Val Ser Pro Ser Gly Gly Phe Val 210 215 220 Ala Glu Leu Leu Arg Pro Glu Leu Asp Pro Val Tyr Ile Pro Pro Gln 225 230 235 240 Gln Pro Gln Pro Pro Gly Gly Gly Leu Met Gly Lys Phe Val Leu Lys 245 250 255 Ala Ser Leu Ser Ala Pro Gly Ser Glu Tyr Gly Ser Pro Ser Val Ile 260 265 270 Ser Val Ser Lys Gly Ser Pro Asp Gly Ser His Pro Val Val Val Ala 275 280 285 Pro Tyr Asn Gly Gly Pro Pro Arg Thr Cys Pro Lys Ile Lys Gln Glu 290 295 300 Ala Val Ser Ser Cys Thr His Leu Gly Ala Gly Pro Pro Leu Ser Asn 305 310 315 320 Gly His Arg Pro Ala Ala His Asp Phe Pro Leu Gly Arg Gln Leu Pro 325 330 335 Ser Arg Thr Thr Pro Thr Leu Gly Leu Glu Glu Val Leu Ser Ser Arg 340 345 350 Asp Cys His Pro Ala Leu Pro Leu Pro Pro Gly Phe His Pro His Pro 355 360 365 Gly Pro Asn Tyr Pro Ser Phe Leu Pro Asp Gln Met Gln Pro Gln Val 370 375 380 Pro Pro Leu His Tyr Gln Glu Leu Met Pro Pro Gly Ser Cys Met Pro 385 390 395 400 Glu Glu Pro Lys Pro Lys Arg Gly Arg Arg Ser Trp Pro Arg Lys Arg 405 410 415 Thr Ala Thr His Thr Cys Asp Tyr Ala Gly Cys Gly Lys Thr Tyr Thr 420 425 430 Lys Ser Ser His Leu Lys Ala His Leu Arg Thr His Thr Gly Glu Lys 435 440 445 Pro Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser 450 455 460 Asp Glu Leu Thr Arg His Tyr Arg Lys His Thr Gly His Arg Pro Phe 465 470 475 480 Gln Cys Gln Lys Cys Asp Arg Ala Phe Ser Arg Ser Asp His Leu Ala 485 490 495 Leu His Met Lys Arg His Phe Leu Ala Val Leu Ala Ala Ala Pro 500 505 510 <210> 131 <211> 1383 <212> DNA <213> Artificial Sequence <220> <223> NLS-cMyc cDNA Sequence <400> 131 atgaagaaga agaggaagct ggattttttt cgggtagtgg aaaaccagca gcctcccgcg 60 acgatgcccc tcaacgttag cttcaccaac aggaactatg acctcgacta cgactcggtg 120 cagccgtatt tctactgcga cgaggaggag aacttctacc agcagcagca gcagagcgag 180 ctgcagcccc cggcgcccag cgaggatatc tggaagaaat tcgagctgct gcccaccccg 240 cccctgtccc ctagccgccg ctccgggctc tgctcgccct cctacgttgc ggtcacaccc 300 ttctcccttc ggggagacaa cgacggcggt ggcgggagct tctccacggc cgaccagctg 360 gagatggtga ccgagctgct gggaggagac atggtgaacc agagtttcat ctgcgacccg 420 gacgacgaga ccttcatcaa aaacatcatc atccaggact gtatgtggag cggcttctcg 480 gccgccgcca agctcgtctc agagaagctg gcctcctacc aggctgcgcg caaagacagc 540 ggcagcccga accccgcccg cggccacagc gtctgctcca cctccagctt gtacctgcag 600 gatctgagcg ccgccgcctc agagtgcatc gacccctcgg tggtcttccc ctaccctctc 660 aacgacagca gctcgcccaa gtcctgcgcc tcgcaagact ccagcgcctt ctctccgtcc 720 tcggattctc tgctctcctc gacggagtcc tccccgcagg gcagccccga gcccctggtg 780 ctccatgagg agacaccgcc caccaccagc agcgactctg aggaggaaca agaagatgag 840 gaagaaatcg atgttgtttc tgtggaaaag aggcaggctc ctggcaaaag gtcagagtct 900 ggatcacctt ctgctggagg ccacagcaaa cctcctcaca gcccactggt cctcaagagg 960 tgccacgtct ccacacatca gcacaactac gcagcgcctc cctccactcg gaaggactat 1020 cctgctgcca agagggtcaa gttggacagt gtcagagtcc tgagacagat cagcaacaac 1080 cgaaaatgca ccagccccag gtcctcggac accgaggaga atgtcaagag gcgaacacac 1140 aacgtcttgg agcgccagag gaggaacgag ctaaaacgga gcttttttgc cctgcgtgac 1200 cagatcccgg agttggaaaa caatgaaaag gcccccaagg tagttatcct taaaaaagcc 1260 acagcataca tcctgtccgt ccaagcagag gagcaaaagc tcatttctga agaggacttg 1320 ttgcggaaac gacgagaaca gttgaaacac aaacttgaac agctacggaa ctcttgtgcg 1380 taa 1383 <210> 132 <211> 460 <212> PRT <213> Artificial Sequence <220> <223> NLS-cMyc Amino Acid Sequence <400> 132 Met Lys Lys Lys Arg Lys Leu Asp Phe Phe Arg Val Val Glu Asn Gln 1 5 10 15 Gln Pro Pro Ala Thr Met Pro Leu Asn Val Ser Phe Thr Asn Arg Asn 20 25 30 Tyr Asp Leu Asp Tyr Asp Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu 35 40 45 Glu Glu Asn Phe Tyr Gln Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro 50 55 60 Ala Pro Ser Glu Asp Ile Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro 65 70 75 80 Pro Leu Ser Pro Ser Arg Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val 85 90 95 Ala Val Thr Pro Phe Ser Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly 100 105 110 Ser Phe Ser Thr Ala Asp Gln Leu Glu Met Val Thr Glu Leu Leu Gly 115 120 125 Gly Asp Met Val Asn Gln Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr 130 135 140 Phe Ile Lys Asn Ile Ile Ile Gln Asp Cys Met Trp Ser Gly Phe Ser 145 150 155 160 Ala Ala Ala Lys Leu Val Ser Glu Lys Leu Ala Ser Tyr Gln Ala Ala 165 170 175 Arg Lys Asp Ser Gly Ser Pro Asn Pro Ala Arg Gly His Ser Val Cys 180 185 190 Ser Thr Ser Ser Leu Tyr Leu Gln Asp Leu Ser Ala Ala Ala Ser Glu 195 200 205 Cys Ile Asp Pro Ser Val Val Phe Pro Tyr Pro Leu Asn Asp Ser Ser 210 215 220 Ser Pro Lys Ser Cys Ala Ser Gln Asp Ser Ser Ala Phe Ser Pro Ser 225 230 235 240 Ser Asp Ser Leu Leu Ser Ser Thr Glu Ser Ser Pro Gln Gly Ser Pro 245 250 255 Glu Pro Leu Val Leu His Glu Glu Thr Pro Pro Thr Thr Ser Ser Asp 260 265 270 Ser Glu Glu Glu Gln Glu Asp Glu Glu Glu Ile Asp Val Val Ser Val 275 280 285 Glu Lys Arg Gln Ala Pro Gly Lys Arg Ser Glu Ser Gly Ser Pro Ser 290 295 300 Ala Gly Gly His Ser Lys Pro Pro His Ser Pro Leu Val Leu Lys Arg 305 310 315 320 Cys His Val Ser Thr His Gln His Asn Tyr Ala Ala Pro Pro Ser Thr 325 330 335 Arg Lys Asp Tyr Pro Ala Ala Lys Arg Val Lys Leu Asp Ser Val Arg 340 345 350 Val Leu Arg Gln Ile Ser Asn Asn Arg Lys Cys Thr Ser Pro Arg Ser 355 360 365 Ser Asp Thr Glu Glu Asn Val Lys Arg Arg Thr His Asn Val Leu Glu 370 375 380 Arg Gln Arg Arg Asn Glu Leu Lys Arg Ser Phe Phe Ala Leu Arg Asp 385 390 395 400 Gln Ile Pro Glu Leu Glu Asn Asn Glu Lys Ala Pro Lys Val Val Ile 405 410 415 Leu Lys Lys Ala Thr Ala Tyr Ile Leu Ser Val Gln Ala Glu Glu Gln 420 425 430 Lys Leu Ile Ser Glu Glu Asp Leu Leu Arg Lys Arg Arg Glu Gln Leu 435 440 445 Lys His Lys Leu Glu Gln Leu Arg Asn Ser Cys Ala 450 455 460 <210> 133 <211> 1410 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-cMyc cDNA Sequence <400> 133 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgctgga tttttttcgg 60 gtagtggaaa accagcagcc tcccgcgacg atgcccctca acgttagctt caccaacagg 120 aactatgacc tcgactacga ctcggtgcag ccgtatttct actgcgacga ggaggagaac 180 ttctaccagc agcagcagca gagcgagctg cagcccccgg cgcccagcga ggatatctgg 240 aagaaattcg agctgctgcc caccccgccc ctgtccccta gccgccgctc cgggctctgc 300 tcgccctcct acgttgcggt cacacccttc tcccttcggg gagacaacga cggcggtggc 360 gggagcttct ccacggccga ccagctggag atggtgaccg agctgctggg aggagacatg 420 gtgaaccaga gtttcatctg cgacccggac gacgagacct tcatcaaaaa catcatcatc 480 caggactgta tgtggagcgg cttctcggcc gccgccaagc tcgtctcaga gaagctggcc 540 tcctaccagg ctgcgcgcaa agacagcggc agcccgaacc ccgcccgcgg ccacagcgtc 600 tgctccacct ccagcttgta cctgcaggat ctgagcgccg ccgcctcaga gtgcatcgac 660 ccctcggtgg tcttccccta ccctctcaac gacagcagct cgcccaagtc ctgcgcctcg 720 caagactcca gcgccttctc tccgtcctcg gattctctgc tctcctcgac ggagtcctcc 780 ccgcagggca gccccgagcc cctggtgctc catgaggaga caccgcccac caccagcagc 840 gactctgagg aggaacaaga agatgaggaa gaaatcgatg ttgtttctgt ggaaaagagg 900 caggctcctg gcaaaaggtc agagtctgga tcaccttctg ctggaggcca cagcaaacct 960 cctcacagcc cactggtcct caagaggtgc cacgtctcca cacatcagca caactacgca 1020 gcgcctccct ccactcggaa ggactatcct gctgccaaga gggtcaagtt ggacagtgtc 1080 agagtcctga gacagatcag caacaaccga aaatgcacca gccccaggtc ctcggacacc 1140 gaggagaatg tcaagaggcg aacacacaac gtcttggagc gccagaggag gaacgagcta 1200 aaacggagct tttttgccct gcgtgaccag atcccggagt tggaaaacaa tgaaaaggcc 1260 cccaaggtag ttatccttaa aaaagccaca gcatacatcc tgtccgtcca agcagaggag 1320 caaaagctca tttctgaaga ggacttgttg cggaaacgac gagaacagtt gaaacacaaa 1380 cttgaacagc tacggaactc ttgtgcgtaa 1410 <210> 134 <211> 469 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-cMyc Amino Acid Sequence <400> 134 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Leu 1 5 10 15 Asp Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro Ala Thr Met Pro 20 25 30 Leu Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu Asp Tyr Asp Ser 35 40 45 Val Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn Phe Tyr Gln Gln 50 55 60 Gln Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser Glu Asp Ile Trp 65 70 75 80 Lys Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser Pro Ser Arg Arg 85 90 95 Ser Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr Pro Phe Ser Leu 100 105 110 Arg Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser Thr Ala Asp Gln 115 120 125 Leu Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met Val Asn Gln Ser 130 135 140 Phe Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys Asn Ile Ile Ile 145 150 155 160 Gln Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala Lys Leu Val Ser 165 170 175 Glu Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp Ser Gly Ser Pro 180 185 190 Asn Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser Ser Leu Tyr Leu 195 200 205 Gln Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp Pro Ser Val Val 210 215 220 Phe Pro Tyr Pro Leu Asn Asp Ser Ser Ser Pro Lys Ser Cys Ala Ser 225 230 235 240 Gln Asp Ser Ser Ala Phe Ser Pro Ser Ser Asp Ser Leu Leu Ser Ser 245 250 255 Thr Glu Ser Ser Pro Gln Gly Ser Pro Glu Pro Leu Val Leu His Glu 260 265 270 Glu Thr Pro Pro Thr Thr Ser Ser Asp Ser Glu Glu Glu Gln Glu Asp 275 280 285 Glu Glu Glu Ile Asp Val Val Ser Val Glu Lys Arg Gln Ala Pro Gly 290 295 300 Lys Arg Ser Glu Ser Gly Ser Pro Ser Ala Gly Gly His Ser Lys Pro 305 310 315 320 Pro His Ser Pro Leu Val Leu Lys Arg Cys His Val Ser Thr His Gln 325 330 335 His Asn Tyr Ala Ala Pro Pro Ser Thr Arg Lys Asp Tyr Pro Ala Ala 340 345 350 Lys Arg Val Lys Leu Asp Ser Val Arg Val Leu Arg Gln Ile Ser Asn 355 360 365 Asn Arg Lys Cys Thr Ser Pro Arg Ser Ser Asp Thr Glu Glu Asn Val 370 375 380 Lys Arg Arg Thr His Asn Val Leu Glu Arg Gln Arg Arg Asn Glu Leu 385 390 395 400 Lys Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro Glu Leu Glu Asn 405 410 415 Asn Glu Lys Ala Pro Lys Val Val Ile Leu Lys Lys Ala Thr Ala Tyr 420 425 430 Ile Leu Ser Val Gln Ala Glu Glu Gln Lys Leu Ile Ser Glu Glu Asp 435 440 445 Leu Leu Arg Lys Arg Arg Glu Gln Leu Lys His Lys Leu Glu Gln Leu 450 455 460 Arg Asn Ser Cys Ala 465 <210> 135 <211> 1410 <212> DNA <213> Artificial Sequence <220> <223> NLS-cMyc-JO-84 MTD cDNA Sequence <400> 135 atgaagaaga agaggaagct ggattttttt cgggtagtgg aaaaccagca gcctcccgcg 60 acgatgcccc tcaacgttag cttcaccaac aggaactatg acctcgacta cgactcggtg 120 cagccgtatt tctactgcga cgaggaggag aacttctacc agcagcagca gcagagcgag 180 ctgcagcccc cggcgcccag cgaggatatc tggaagaaat tcgagctgct gcccaccccg 240 cccctgtccc ctagccgccg ctccgggctc tgctcgccct cctacgttgc ggtcacaccc 300 ttctcccttc ggggagacaa cgacggcggt ggcgggagct tctccacggc cgaccagctg 360 gagatggtga ccgagctgct gggaggagac atggtgaacc agagtttcat ctgcgacccg 420 gacgacgaga ccttcatcaa aaacatcatc atccaggact gtatgtggag cggcttctcg 480 gccgccgcca agctcgtctc agagaagctg gcctcctacc aggctgcgcg caaagacagc 540 ggcagcccga accccgcccg cggccacagc gtctgctcca cctccagctt gtacctgcag 600 gatctgagcg ccgccgcctc agagtgcatc gacccctcgg tggtcttccc ctaccctctc 660 aacgacagca gctcgcccaa gtcctgcgcc tcgcaagact ccagcgcctt ctctccgtcc 720 tcggattctc tgctctcctc gacggagtcc tccccgcagg gcagccccga gcccctggtg 780 ctccatgagg agacaccgcc caccaccagc agcgactctg aggaggaaca agaagatgag 840 gaagaaatcg atgttgtttc tgtggaaaag aggcaggctc ctggcaaaag gtcagagtct 900 ggatcacctt ctgctggagg ccacagcaaa cctcctcaca gcccactggt cctcaagagg 960 tgccacgtct ccacacatca gcacaactac gcagcgcctc cctccactcg gaaggactat 1020 cctgctgcca agagggtcaa gttggacagt gtcagagtcc tgagacagat cagcaacaac 1080 cgaaaatgca ccagccccag gtcctcggac accgaggaga atgtcaagag gcgaacacac 1140 aacgtcttgg agcgccagag gaggaacgag ctaaaacgga gcttttttgc cctgcgtgac 1200 cagatcccgg agttggaaaa caatgaaaag gcccccaagg tagttatcct taaaaaagcc 1260 acagcataca tcctgtccgt ccaagcagag gagcaaaagc tcatttctga agaggacttg 1320 ttgcggaaac gacgagaaca gttgaaacac aaacttgaac agctacggaa ctcttgtgcg 1380 ctggtggcgg cgctgctggc ggtgctgtaa 1410 <210> 136 <211> 469 <212> PRT <213> Artificial Sequence <220> <223> NLS-cMyc-JO-84 MTD Amino Acid Sequence <400> 136 Met Lys Lys Lys Arg Lys Leu Asp Phe Phe Arg Val Val Glu Asn Gln 1 5 10 15 Gln Pro Pro Ala Thr Met Pro Leu Asn Val Ser Phe Thr Asn Arg Asn 20 25 30 Tyr Asp Leu Asp Tyr Asp Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu 35 40 45 Glu Glu Asn Phe Tyr Gln Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro 50 55 60 Ala Pro Ser Glu Asp Ile Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro 65 70 75 80 Pro Leu Ser Pro Ser Arg Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val 85 90 95 Ala Val Thr Pro Phe Ser Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly 100 105 110 Ser Phe Ser Thr Ala Asp Gln Leu Glu Met Val Thr Glu Leu Leu Gly 115 120 125 Gly Asp Met Val Asn Gln Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr 130 135 140 Phe Ile Lys Asn Ile Ile Ile Gln Asp Cys Met Trp Ser Gly Phe Ser 145 150 155 160 Ala Ala Ala Lys Leu Val Ser Glu Lys Leu Ala Ser Tyr Gln Ala Ala 165 170 175 Arg Lys Asp Ser Gly Ser Pro Asn Pro Ala Arg Gly His Ser Val Cys 180 185 190 Ser Thr Ser Ser Leu Tyr Leu Gln Asp Leu Ser Ala Ala Ala Ser Glu 195 200 205 Cys Ile Asp Pro Ser Val Val Phe Pro Tyr Pro Leu Asn Asp Ser Ser 210 215 220 Ser Pro Lys Ser Cys Ala Ser Gln Asp Ser Ser Ala Phe Ser Pro Ser 225 230 235 240 Ser Asp Ser Leu Leu Ser Ser Thr Glu Ser Ser Pro Gln Gly Ser Pro 245 250 255 Glu Pro Leu Val Leu His Glu Glu Thr Pro Pro Thr Thr Ser Ser Asp 260 265 270 Ser Glu Glu Glu Gln Glu Asp Glu Glu Glu Ile Asp Val Val Ser Val 275 280 285 Glu Lys Arg Gln Ala Pro Gly Lys Arg Ser Glu Ser Gly Ser Pro Ser 290 295 300 Ala Gly Gly His Ser Lys Pro Pro His Ser Pro Leu Val Leu Lys Arg 305 310 315 320 Cys His Val Ser Thr His Gln His Asn Tyr Ala Ala Pro Pro Ser Thr 325 330 335 Arg Lys Asp Tyr Pro Ala Ala Lys Arg Val Lys Leu Asp Ser Val Arg 340 345 350 Val Leu Arg Gln Ile Ser Asn Asn Arg Lys Cys Thr Ser Pro Arg Ser 355 360 365 Ser Asp Thr Glu Glu Asn Val Lys Arg Arg Thr His Asn Val Leu Glu 370 375 380 Arg Gln Arg Arg Asn Glu Leu Lys Arg Ser Phe Phe Ala Leu Arg Asp 385 390 395 400 Gln Ile Pro Glu Leu Glu Asn Asn Glu Lys Ala Pro Lys Val Val Ile 405 410 415 Leu Lys Lys Ala Thr Ala Tyr Ile Leu Ser Val Gln Ala Glu Glu Gln 420 425 430 Lys Leu Ile Ser Glu Glu Asp Leu Leu Arg Lys Arg Arg Glu Gln Leu 435 440 445 Lys His Lys Leu Glu Gln Leu Arg Asn Ser Cys Ala Leu Val Ala Ala 450 455 460 Leu Leu Ala Val Leu 465 <210> 137 <211> 1437 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-cMyc-JO-84 MTD cDNA Sequence <400> 137 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgctgga tttttttcgg 60 gtagtggaaa accagcagcc tcccgcgacg atgcccctca acgttagctt caccaacagg 120 aactatgacc tcgactacga ctcggtgcag ccgtatttct actgcgacga ggaggagaac 180 ttctaccagc agcagcagca gagcgagctg cagcccccgg cgcccagcga ggatatctgg 240 aagaaattcg agctgctgcc caccccgccc ctgtccccta gccgccgctc cgggctctgc 300 tcgccctcct acgttgcggt cacacccttc tcccttcggg gagacaacga cggcggtggc 360 gggagcttct ccacggccga ccagctggag atggtgaccg agctgctggg aggagacatg 420 gtgaaccaga gtttcatctg cgacccggac gacgagacct tcatcaaaaa catcatcatc 480 caggactgta tgtggagcgg cttctcggcc gccgccaagc tcgtctcaga gaagctggcc 540 tcctaccagg ctgcgcgcaa agacagcggc agcccgaacc ccgcccgcgg ccacagcgtc 600 tgctccacct ccagcttgta cctgcaggat ctgagcgccg ccgcctcaga gtgcatcgac 660 ccctcggtgg tcttccccta ccctctcaac gacagcagct cgcccaagtc ctgcgcctcg 720 caagactcca gcgccttctc tccgtcctcg gattctctgc tctcctcgac ggagtcctcc 780 ccgcagggca gccccgagcc cctggtgctc catgaggaga caccgcccac caccagcagc 840 gactctgagg aggaacaaga agatgaggaa gaaatcgatg ttgtttctgt ggaaaagagg 900 caggctcctg gcaaaaggtc agagtctgga tcaccttctg ctggaggcca cagcaaacct 960 cctcacagcc cactggtcct caagaggtgc cacgtctcca cacatcagca caactacgca 1020 gcgcctccct ccactcggaa ggactatcct gctgccaaga gggtcaagtt ggacagtgtc 1080 agagtcctga gacagatcag caacaaccga aaatgcacca gccccaggtc ctcggacacc 1140 gaggagaatg tcaagaggcg aacacacaac gtcttggagc gccagaggag gaacgagcta 1200 aaacggagct tttttgccct gcgtgaccag atcccggagt tggaaaacaa tgaaaaggcc 1260 cccaaggtag ttatccttaa aaaagccaca gcatacatcc tgtccgtcca agcagaggag 1320 caaaagctca tttctgaaga ggacttgttg cggaaacgac gagaacagtt gaaacacaaa 1380 cttgaacagc tacggaactc ttgtgcgctg gtggcggcgc tgctggcggt gctgtaa 1437 <210> 138 <211> 478 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-cMyc-JO-84 MTD Amino Acid Sequence <400> 138 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Leu 1 5 10 15 Asp Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro Ala Thr Met Pro 20 25 30 Leu Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu Asp Tyr Asp Ser 35 40 45 Val Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn Phe Tyr Gln Gln 50 55 60 Gln Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser Glu Asp Ile Trp 65 70 75 80 Lys Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser Pro Ser Arg Arg 85 90 95 Ser Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr Pro Phe Ser Leu 100 105 110 Arg Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser Thr Ala Asp Gln 115 120 125 Leu Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met Val Asn Gln Ser 130 135 140 Phe Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys Asn Ile Ile Ile 145 150 155 160 Gln Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala Lys Leu Val Ser 165 170 175 Glu Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp Ser Gly Ser Pro 180 185 190 Asn Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser Ser Leu Tyr Leu 195 200 205 Gln Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp Pro Ser Val Val 210 215 220 Phe Pro Tyr Pro Leu Asn Asp Ser Ser Ser Pro Lys Ser Cys Ala Ser 225 230 235 240 Gln Asp Ser Ser Ala Phe Ser Pro Ser Ser Asp Ser Leu Leu Ser Ser 245 250 255 Thr Glu Ser Ser Pro Gln Gly Ser Pro Glu Pro Leu Val Leu His Glu 260 265 270 Glu Thr Pro Pro Thr Thr Ser Ser Asp Ser Glu Glu Glu Gln Glu Asp 275 280 285 Glu Glu Glu Ile Asp Val Val Ser Val Glu Lys Arg Gln Ala Pro Gly 290 295 300 Lys Arg Ser Glu Ser Gly Ser Pro Ser Ala Gly Gly His Ser Lys Pro 305 310 315 320 Pro His Ser Pro Leu Val Leu Lys Arg Cys His Val Ser Thr His Gln 325 330 335 His Asn Tyr Ala Ala Pro Pro Ser Thr Arg Lys Asp Tyr Pro Ala Ala 340 345 350 Lys Arg Val Lys Leu Asp Ser Val Arg Val Leu Arg Gln Ile Ser Asn 355 360 365 Asn Arg Lys Cys Thr Ser Pro Arg Ser Ser Asp Thr Glu Glu Asn Val 370 375 380 Lys Arg Arg Thr His Asn Val Leu Glu Arg Gln Arg Arg Asn Glu Leu 385 390 395 400 Lys Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro Glu Leu Glu Asn 405 410 415 Asn Glu Lys Ala Pro Lys Val Val Ile Leu Lys Lys Ala Thr Ala Tyr 420 425 430 Ile Leu Ser Val Gln Ala Glu Glu Gln Lys Leu Ile Ser Glu Glu Asp 435 440 445 Leu Leu Arg Lys Arg Arg Glu Gln Leu Lys His Lys Leu Glu Gln Leu 450 455 460 Arg Asn Ser Cys Ala Leu Val Ala Ala Leu Leu Ala Val Leu 465 470 475 <210> 139 <211> 1407 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-cMyc cDNA Sequence <400> 139 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgctggattt ttttcgggta 60 gtggaaaacc agcagcctcc cgcgacgatg cccctcaacg ttagcttcac caacaggaac 120 tatgacctcg actacgactc ggtgcagccg tatttctact gcgacgagga ggagaacttc 180 taccagcagc agcagcagag cgagctgcag cccccggcgc ccagcgagga tatctggaag 240 aaattcgagc tgctgcccac cccgcccctg tcccctagcc gccgctccgg gctctgctcg 300 ccctcctacg ttgcggtcac acccttctcc cttcggggag acaacgacgg cggtggcggg 360 agcttctcca cggccgacca gctggagatg gtgaccgagc tgctgggagg agacatggtg 420 aaccagagtt tcatctgcga cccggacgac gagaccttca tcaaaaacat catcatccag 480 gactgtatgt ggagcggctt ctcggccgcc gccaagctcg tctcagagaa gctggcctcc 540 taccaggctg cgcgcaaaga cagcggcagc ccgaaccccg cccgcggcca cagcgtctgc 600 tccacctcca gcttgtacct gcaggatctg agcgccgccg cctcagagtg catcgacccc 660 tcggtggtct tcccctaccc tctcaacgac agcagctcgc ccaagtcctg cgcctcgcaa 720 gactccagcg ccttctctcc gtcctcggat tctctgctct cctcgacgga gtcctccccg 780 cagggcagcc ccgagcccct ggtgctccat gaggagacac cgcccaccac cagcagcgac 840 tctgaggagg aacaagaaga tgaggaagaa atcgatgttg tttctgtgga aaagaggcag 900 gctcctggca aaaggtcaga gtctggatca ccttctgctg gaggccacag caaacctcct 960 cacagcccac tggtcctcaa gaggtgccac gtctccacac atcagcacaa ctacgcagcg 1020 cctccctcca ctcggaagga ctatcctgct gccaagaggg tcaagttgga cagtgtcaga 1080 gtcctgagac agatcagcaa caaccgaaaa tgcaccagcc ccaggtcctc ggacaccgag 1140 gagaatgtca agaggcgaac acacaacgtc ttggagcgcc agaggaggaa cgagctaaaa 1200 cggagctttt ttgccctgcg tgaccagatc ccggagttgg aaaacaatga aaaggccccc 1260 aaggtagtta tccttaaaaa agccacagca tacatcctgt ccgtccaagc agaggagcaa 1320 aagctcattt ctgaagagga cttgttgcgg aaacgacgag aacagttgaa acacaaactt 1380 gaacagctac ggaactcttg tgcgtaa 1407 <210> 140 <211> 468 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-cMyc Amino Acid Sequence <400> 140 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Leu Asp 1 5 10 15 Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro Ala Thr Met Pro Leu 20 25 30 Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu Asp Tyr Asp Ser Val 35 40 45 Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn Phe Tyr Gln Gln Gln 50 55 60 Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser Glu Asp Ile Trp Lys 65 70 75 80 Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser Pro Ser Arg Arg Ser 85 90 95 Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr Pro Phe Ser Leu Arg 100 105 110 Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser Thr Ala Asp Gln Leu 115 120 125 Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met Val Asn Gln Ser Phe 130 135 140 Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys Asn Ile Ile Ile Gln 145 150 155 160 Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala Lys Leu Val Ser Glu 165 170 175 Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp Ser Gly Ser Pro Asn 180 185 190 Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser Ser Leu Tyr Leu Gln 195 200 205 Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp Pro Ser Val Val Phe 210 215 220 Pro Tyr Pro Leu Asn Asp Ser Ser Ser Pro Lys Ser Cys Ala Ser Gln 225 230 235 240 Asp Ser Ser Ala Phe Ser Pro Ser Ser Asp Ser Leu Leu Ser Ser Thr 245 250 255 Glu Ser Ser Pro Gln Gly Ser Pro Glu Pro Leu Val Leu His Glu Glu 260 265 270 Thr Pro Pro Thr Thr Ser Ser Asp Ser Glu Glu Glu Gln Glu Asp Glu 275 280 285 Glu Glu Ile Asp Val Val Ser Val Glu Lys Arg Gln Ala Pro Gly Lys 290 295 300 Arg Ser Glu Ser Gly Ser Pro Ser Ala Gly Gly His Ser Lys Pro Pro 305 310 315 320 His Ser Pro Leu Val Leu Lys Arg Cys His Val Ser Thr His Gln His 325 330 335 Asn Tyr Ala Ala Pro Pro Ser Thr Arg Lys Asp Tyr Pro Ala Ala Lys 340 345 350 Arg Val Lys Leu Asp Ser Val Arg Val Leu Arg Gln Ile Ser Asn Asn 355 360 365 Arg Lys Cys Thr Ser Pro Arg Ser Ser Asp Thr Glu Glu Asn Val Lys 370 375 380 Arg Arg Thr His Asn Val Leu Glu Arg Gln Arg Arg Asn Glu Leu Lys 385 390 395 400 Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro Glu Leu Glu Asn Asn 405 410 415 Glu Lys Ala Pro Lys Val Val Ile Leu Lys Lys Ala Thr Ala Tyr Ile 420 425 430 Leu Ser Val Gln Ala Glu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 435 440 445 Leu Arg Lys Arg Arg Glu Gln Leu Lys His Lys Leu Glu Gln Leu Arg 450 455 460 Asn Ser Cys Ala 465 <210> 141 <211> 1407 <212> DNA <213> Artificial Sequence <220> <223> NLS-cMyc-JO-86 MTD cDNA Sequence <400> 141 atgaagaaga agaggaagct ggattttttt cgggtagtgg aaaaccagca gcctcccgcg 60 acgatgcccc tcaacgttag cttcaccaac aggaactatg acctcgacta cgactcggtg 120 cagccgtatt tctactgcga cgaggaggag aacttctacc agcagcagca gcagagcgag 180 ctgcagcccc cggcgcccag cgaggatatc tggaagaaat tcgagctgct gcccaccccg 240 cccctgtccc ctagccgccg ctccgggctc tgctcgccct cctacgttgc ggtcacaccc 300 ttctcccttc ggggagacaa cgacggcggt ggcgggagct tctccacggc cgaccagctg 360 gagatggtga ccgagctgct gggaggagac atggtgaacc agagtttcat ctgcgacccg 420 gacgacgaga ccttcatcaa aaacatcatc atccaggact gtatgtggag cggcttctcg 480 gccgccgcca agctcgtctc agagaagctg gcctcctacc aggctgcgcg caaagacagc 540 ggcagcccga accccgcccg cggccacagc gtctgctcca cctccagctt gtacctgcag 600 gatctgagcg ccgccgcctc agagtgcatc gacccctcgg tggtcttccc ctaccctctc 660 aacgacagca gctcgcccaa gtcctgcgcc tcgcaagact ccagcgcctt ctctccgtcc 720 tcggattctc tgctctcctc gacggagtcc tccccgcagg gcagccccga gcccctggtg 780 ctccatgagg agacaccgcc caccaccagc agcgactctg aggaggaaca agaagatgag 840 gaagaaatcg atgttgtttc tgtggaaaag aggcaggctc ctggcaaaag gtcagagtct 900 ggatcacctt ctgctggagg ccacagcaaa cctcctcaca gcccactggt cctcaagagg 960 tgccacgtct ccacacatca gcacaactac gcagcgcctc cctccactcg gaaggactat 1020 cctgctgcca agagggtcaa gttggacagt gtcagagtcc tgagacagat cagcaacaac 1080 cgaaaatgca ccagccccag gtcctcggac accgaggaga atgtcaagag gcgaacacac 1140 aacgtcttgg agcgccagag gaggaacgag ctaaaacgga gcttttttgc cctgcgtgac 1200 cagatcccgg agttggaaaa caatgaaaag gcccccaagg tagttatcct taaaaaagcc 1260 acagcataca tcctgtccgt ccaagcagag gagcaaaagc tcatttctga agaggacttg 1320 ttgcggaaac gacgagaaca gttgaaacac aaacttgaac agctacggaa ctcttgtgcg 1380 ctggcggtgc tggcggcggc gccgtaa 1407 <210> 142 <211> 468 <212> PRT <213> Artificial Sequence <220> <223> NLS-cMyc-JO-86 MTD Amino Acid Sequence <400> 142 Met Lys Lys Lys Arg Lys Leu Asp Phe Phe Arg Val Val Glu Asn Gln 1 5 10 15 Gln Pro Pro Ala Thr Met Pro Leu Asn Val Ser Phe Thr Asn Arg Asn 20 25 30 Tyr Asp Leu Asp Tyr Asp Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu 35 40 45 Glu Glu Asn Phe Tyr Gln Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro 50 55 60 Ala Pro Ser Glu Asp Ile Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro 65 70 75 80 Pro Leu Ser Pro Ser Arg Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val 85 90 95 Ala Val Thr Pro Phe Ser Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly 100 105 110 Ser Phe Ser Thr Ala Asp Gln Leu Glu Met Val Thr Glu Leu Leu Gly 115 120 125 Gly Asp Met Val Asn Gln Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr 130 135 140 Phe Ile Lys Asn Ile Ile Ile Gln Asp Cys Met Trp Ser Gly Phe Ser 145 150 155 160 Ala Ala Ala Lys Leu Val Ser Glu Lys Leu Ala Ser Tyr Gln Ala Ala 165 170 175 Arg Lys Asp Ser Gly Ser Pro Asn Pro Ala Arg Gly His Ser Val Cys 180 185 190 Ser Thr Ser Ser Leu Tyr Leu Gln Asp Leu Ser Ala Ala Ala Ser Glu 195 200 205 Cys Ile Asp Pro Ser Val Val Phe Pro Tyr Pro Leu Asn Asp Ser Ser 210 215 220 Ser Pro Lys Ser Cys Ala Ser Gln Asp Ser Ser Ala Phe Ser Pro Ser 225 230 235 240 Ser Asp Ser Leu Leu Ser Ser Thr Glu Ser Ser Pro Gln Gly Ser Pro 245 250 255 Glu Pro Leu Val Leu His Glu Glu Thr Pro Pro Thr Thr Ser Ser Asp 260 265 270 Ser Glu Glu Glu Gln Glu Asp Glu Glu Glu Ile Asp Val Val Ser Val 275 280 285 Glu Lys Arg Gln Ala Pro Gly Lys Arg Ser Glu Ser Gly Ser Pro Ser 290 295 300 Ala Gly Gly His Ser Lys Pro Pro His Ser Pro Leu Val Leu Lys Arg 305 310 315 320 Cys His Val Ser Thr His Gln His Asn Tyr Ala Ala Pro Pro Ser Thr 325 330 335 Arg Lys Asp Tyr Pro Ala Ala Lys Arg Val Lys Leu Asp Ser Val Arg 340 345 350 Val Leu Arg Gln Ile Ser Asn Asn Arg Lys Cys Thr Ser Pro Arg Ser 355 360 365 Ser Asp Thr Glu Glu Asn Val Lys Arg Arg Thr His Asn Val Leu Glu 370 375 380 Arg Gln Arg Arg Asn Glu Leu Lys Arg Ser Phe Phe Ala Leu Arg Asp 385 390 395 400 Gln Ile Pro Glu Leu Glu Asn Asn Glu Lys Ala Pro Lys Val Val Ile 405 410 415 Leu Lys Lys Ala Thr Ala Tyr Ile Leu Ser Val Gln Ala Glu Glu Gln 420 425 430 Lys Leu Ile Ser Glu Glu Asp Leu Leu Arg Lys Arg Arg Glu Gln Leu 435 440 445 Lys His Lys Leu Glu Gln Leu Arg Asn Ser Cys Ala Leu Ala Val Leu 450 455 460 Ala Ala Ala Pro 465 <210> 143 <211> 1431 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-cMyc-JO-86 MTD cDNA Sequence <400> 143 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgctggattt ttttcgggta 60 gtggaaaacc agcagcctcc cgcgacgatg cccctcaacg ttagcttcac caacaggaac 120 tatgacctcg actacgactc ggtgcagccg tatttctact gcgacgagga ggagaacttc 180 taccagcagc agcagcagag cgagctgcag cccccggcgc ccagcgagga tatctggaag 240 aaattcgagc tgctgcccac cccgcccctg tcccctagcc gccgctccgg gctctgctcg 300 ccctcctacg ttgcggtcac acccttctcc cttcggggag acaacgacgg cggtggcggg 360 agcttctcca cggccgacca gctggagatg gtgaccgagc tgctgggagg agacatggtg 420 aaccagagtt tcatctgcga cccggacgac gagaccttca tcaaaaacat catcatccag 480 gactgtatgt ggagcggctt ctcggccgcc gccaagctcg tctcagagaa gctggcctcc 540 taccaggctg cgcgcaaaga cagcggcagc ccgaaccccg cccgcggcca cagcgtctgc 600 tccacctcca gcttgtacct gcaggatctg agcgccgccg cctcagagtg catcgacccc 660 tcggtggtct tcccctaccc tctcaacgac agcagctcgc ccaagtcctg cgcctcgcaa 720 gactccagcg ccttctctcc gtcctcggat tctctgctct cctcgacgga gtcctccccg 780 cagggcagcc ccgagcccct ggtgctccat gaggagacac cgcccaccac cagcagcgac 840 tctgaggagg aacaagaaga tgaggaagaa atcgatgttg tttctgtgga aaagaggcag 900 gctcctggca aaaggtcaga gtctggatca ccttctgctg gaggccacag caaacctcct 960 cacagcccac tggtcctcaa gaggtgccac gtctccacac atcagcacaa ctacgcagcg 1020 cctccctcca ctcggaagga ctatcctgct gccaagaggg tcaagttgga cagtgtcaga 1080 gtcctgagac agatcagcaa caaccgaaaa tgcaccagcc ccaggtcctc ggacaccgag 1140 gagaatgtca agaggcgaac acacaacgtc ttggagcgcc agaggaggaa cgagctaaaa 1200 cggagctttt ttgccctgcg tgaccagatc ccggagttgg aaaacaatga aaaggccccc 1260 aaggtagtta tccttaaaaa agccacagca tacatcctgt ccgtccaagc agaggagcaa 1320 aagctcattt ctgaagagga cttgttgcgg aaacgacgag aacagttgaa acacaaactt 1380 gaacagctac ggaactcttg tgcgctggcg gtgctggcgg cggcgccgta a 1431 <210> 144 <211> 476 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-cMyc-JO-86 MTD Amino Acid Sequence <400> 144 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Leu Asp 1 5 10 15 Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro Ala Thr Met Pro Leu 20 25 30 Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu Asp Tyr Asp Ser Val 35 40 45 Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn Phe Tyr Gln Gln Gln 50 55 60 Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser Glu Asp Ile Trp Lys 65 70 75 80 Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser Pro Ser Arg Arg Ser 85 90 95 Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr Pro Phe Ser Leu Arg 100 105 110 Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser Thr Ala Asp Gln Leu 115 120 125 Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met Val Asn Gln Ser Phe 130 135 140 Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys Asn Ile Ile Ile Gln 145 150 155 160 Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala Lys Leu Val Ser Glu 165 170 175 Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp Ser Gly Ser Pro Asn 180 185 190 Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser Ser Leu Tyr Leu Gln 195 200 205 Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp Pro Ser Val Val Phe 210 215 220 Pro Tyr Pro Leu Asn Asp Ser Ser Ser Pro Lys Ser Cys Ala Ser Gln 225 230 235 240 Asp Ser Ser Ala Phe Ser Pro Ser Ser Asp Ser Leu Leu Ser Ser Thr 245 250 255 Glu Ser Ser Pro Gln Gly Ser Pro Glu Pro Leu Val Leu His Glu Glu 260 265 270 Thr Pro Pro Thr Thr Ser Ser Asp Ser Glu Glu Glu Gln Glu Asp Glu 275 280 285 Glu Glu Ile Asp Val Val Ser Val Glu Lys Arg Gln Ala Pro Gly Lys 290 295 300 Arg Ser Glu Ser Gly Ser Pro Ser Ala Gly Gly His Ser Lys Pro Pro 305 310 315 320 His Ser Pro Leu Val Leu Lys Arg Cys His Val Ser Thr His Gln His 325 330 335 Asn Tyr Ala Ala Pro Pro Ser Thr Arg Lys Asp Tyr Pro Ala Ala Lys 340 345 350 Arg Val Lys Leu Asp Ser Val Arg Val Leu Arg Gln Ile Ser Asn Asn 355 360 365 Arg Lys Cys Thr Ser Pro Arg Ser Ser Asp Thr Glu Glu Asn Val Lys 370 375 380 Arg Arg Thr His Asn Val Leu Glu Arg Gln Arg Arg Asn Glu Leu Lys 385 390 395 400 Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro Glu Leu Glu Asn Asn 405 410 415 Glu Lys Ala Pro Lys Val Val Ile Leu Lys Lys Ala Thr Ala Tyr Ile 420 425 430 Leu Ser Val Gln Ala Glu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 435 440 445 Leu Arg Lys Arg Arg Glu Gln Leu Lys His Lys Leu Glu Gln Leu Arg 450 455 460 Asn Ser Cys Ala Leu Ala Val Leu Ala Ala Ala Pro 465 470 475 <210> 145 <211> 1443 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-cMyc cDNA Sequence <400> 145 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggattttttt cgggtagtgg aaaaccagca gcctcccgcg 120 acgatgcccc tcaacgttag cttcaccaac aggaactatg acctcgacta cgactcggtg 180 cagccgtatt tctactgcga cgaggaggag aacttctacc agcagcagca gcagagcgag 240 ctgcagcccc cggcgcccag cgaggatatc tggaagaaat tcgagctgct gcccaccccg 300 cccctgtccc ctagccgccg ctccgggctc tgctcgccct cctacgttgc ggtcacaccc 360 ttctcccttc ggggagacaa cgacggcggt ggcgggagct tctccacggc cgaccagctg 420 gagatggtga ccgagctgct gggaggagac atggtgaacc agagtttcat ctgcgacccg 480 gacgacgaga ccttcatcaa aaacatcatc atccaggact gtatgtggag cggcttctcg 540 gccgccgcca agctcgtctc agagaagctg gcctcctacc aggctgcgcg caaagacagc 600 ggcagcccga accccgcccg cggccacagc gtctgctcca cctccagctt gtacctgcag 660 gatctgagcg ccgccgcctc agagtgcatc gacccctcgg tggtcttccc ctaccctctc 720 aacgacagca gctcgcccaa gtcctgcgcc tcgcaagact ccagcgcctt ctctccgtcc 780 tcggattctc tgctctcctc gacggagtcc tccccgcagg gcagccccga gcccctggtg 840 ctccatgagg agacaccgcc caccaccagc agcgactctg aggaggaaca agaagatgag 900 gaagaaatcg atgttgtttc tgtggaaaag aggcaggctc ctggcaaaag gtcagagtct 960 ggatcacctt ctgctggagg ccacagcaaa cctcctcaca gcccactggt cctcaagagg 1020 tgccacgtct ccacacatca gcacaactac gcagcgcctc cctccactcg gaaggactat 1080 cctgctgcca agagggtcaa gttggacagt gtcagagtcc tgagacagat cagcaacaac 1140 cgaaaatgca ccagccccag gtcctcggac accgaggaga atgtcaagag gcgaacacac 1200 aacgtcttgg agcgccagag gaggaacgag ctaaaacgga gcttttttgc cctgcgtgac 1260 cagatcccgg agttggaaaa caatgaaaag gcccccaagg tagttatcct taaaaaagcc 1320 acagcataca tcctgtccgt ccaagcagag gagcaaaagc tcatttctga agaggacttg 1380 ttgcggaaac gacgagaaca gttgaaacac aaacttgaac agctacggaa ctcttgtgcg 1440 taa 1443 <210> 146 <211> 480 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-cMyc Amino Acid Sequence <400> 146 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Asp Phe Phe Arg Val 20 25 30 Val Glu Asn Gln Gln Pro Pro Ala Thr Met Pro Leu Asn Val Ser Phe 35 40 45 Thr Asn Arg Asn Tyr Asp Leu Asp Tyr Asp Ser Val Gln Pro Tyr Phe 50 55 60 Tyr Cys Asp Glu Glu Glu Asn Phe Tyr Gln Gln Gln Gln Gln Ser Glu 65 70 75 80 Leu Gln Pro Pro Ala Pro Ser Glu Asp Ile Trp Lys Lys Phe Glu Leu 85 90 95 Leu Pro Thr Pro Pro Leu Ser Pro Ser Arg Arg Ser Gly Leu Cys Ser 100 105 110 Pro Ser Tyr Val Ala Val Thr Pro Phe Ser Leu Arg Gly Asp Asn Asp 115 120 125 Gly Gly Gly Gly Ser Phe Ser Thr Ala Asp Gln Leu Glu Met Val Thr 130 135 140 Glu Leu Leu Gly Gly Asp Met Val Asn Gln Ser Phe Ile Cys Asp Pro 145 150 155 160 Asp Asp Glu Thr Phe Ile Lys Asn Ile Ile Ile Gln Asp Cys Met Trp 165 170 175 Ser Gly Phe Ser Ala Ala Ala Lys Leu Val Ser Glu Lys Leu Ala Ser 180 185 190 Tyr Gln Ala Ala Arg Lys Asp Ser Gly Ser Pro Asn Pro Ala Arg Gly 195 200 205 His Ser Val Cys Ser Thr Ser Ser Leu Tyr Leu Gln Asp Leu Ser Ala 210 215 220 Ala Ala Ser Glu Cys Ile Asp Pro Ser Val Val Phe Pro Tyr Pro Leu 225 230 235 240 Asn Asp Ser Ser Ser Pro Lys Ser Cys Ala Ser Gln Asp Ser Ser Ala 245 250 255 Phe Ser Pro Ser Ser Asp Ser Leu Leu Ser Ser Thr Glu Ser Ser Pro 260 265 270 Gln Gly Ser Pro Glu Pro Leu Val Leu His Glu Glu Thr Pro Pro Thr 275 280 285 Thr Ser Ser Asp Ser Glu Glu Glu Gln Glu Asp Glu Glu Glu Ile Asp 290 295 300 Val Val Ser Val Glu Lys Arg Gln Ala Pro Gly Lys Arg Ser Glu Ser 305 310 315 320 Gly Ser Pro Ser Ala Gly Gly His Ser Lys Pro Pro His Ser Pro Leu 325 330 335 Val Leu Lys Arg Cys His Val Ser Thr His Gln His Asn Tyr Ala Ala 340 345 350 Pro Pro Ser Thr Arg Lys Asp Tyr Pro Ala Ala Lys Arg Val Lys Leu 355 360 365 Asp Ser Val Arg Val Leu Arg Gln Ile Ser Asn Asn Arg Lys Cys Thr 370 375 380 Ser Pro Arg Ser Ser Asp Thr Glu Glu Asn Val Lys Arg Arg Thr His 385 390 395 400 Asn Val Leu Glu Arg Gln Arg Arg Asn Glu Leu Lys Arg Ser Phe Phe 405 410 415 Ala Leu Arg Asp Gln Ile Pro Glu Leu Glu Asn Asn Glu Lys Ala Pro 420 425 430 Lys Val Val Ile Leu Lys Lys Ala Thr Ala Tyr Ile Leu Ser Val Gln 435 440 445 Ala Glu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Leu Arg Lys Arg 450 455 460 Arg Glu Gln Leu Lys His Lys Leu Glu Gln Leu Arg Asn Ser Cys Ala 465 470 475 480 <210> 147 <211> 1470 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-cMyc cDNA Sequence <400> 147 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgctgga tttttttcgg 120 gtagtggaaa accagcagcc tcccgcgacg atgcccctca acgttagctt caccaacagg 180 aactatgacc tcgactacga ctcggtgcag ccgtatttct actgcgacga ggaggagaac 240 ttctaccagc agcagcagca gagcgagctg cagcccccgg cgcccagcga ggatatctgg 300 aagaaattcg agctgctgcc caccccgccc ctgtccccta gccgccgctc cgggctctgc 360 tcgccctcct acgttgcggt cacacccttc tcccttcggg gagacaacga cggcggtggc 420 gggagcttct ccacggccga ccagctggag atggtgaccg agctgctggg aggagacatg 480 gtgaaccaga gtttcatctg cgacccggac gacgagacct tcatcaaaaa catcatcatc 540 caggactgta tgtggagcgg cttctcggcc gccgccaagc tcgtctcaga gaagctggcc 600 tcctaccagg ctgcgcgcaa agacagcggc agcccgaacc ccgcccgcgg ccacagcgtc 660 tgctccacct ccagcttgta cctgcaggat ctgagcgccg ccgcctcaga gtgcatcgac 720 ccctcggtgg tcttccccta ccctctcaac gacagcagct cgcccaagtc ctgcgcctcg 780 caagactcca gcgccttctc tccgtcctcg gattctctgc tctcctcgac ggagtcctcc 840 ccgcagggca gccccgagcc cctggtgctc catgaggaga caccgcccac caccagcagc 900 gactctgagg aggaacaaga agatgaggaa gaaatcgatg ttgtttctgt ggaaaagagg 960 caggctcctg gcaaaaggtc agagtctgga tcaccttctg ctggaggcca cagcaaacct 1020 cctcacagcc cactggtcct caagaggtgc cacgtctcca cacatcagca caactacgca 1080 gcgcctccct ccactcggaa ggactatcct gctgccaaga gggtcaagtt ggacagtgtc 1140 agagtcctga gacagatcag caacaaccga aaatgcacca gccccaggtc ctcggacacc 1200 gaggagaatg tcaagaggcg aacacacaac gtcttggagc gccagaggag gaacgagcta 1260 aaacggagct tttttgccct gcgtgaccag atcccggagt tggaaaacaa tgaaaaggcc 1320 cccaaggtag ttatccttaa aaaagccaca gcatacatcc tgtccgtcca agcagaggag 1380 caaaagctca tttctgaaga ggacttgttg cggaaacgac gagaacagtt gaaacacaaa 1440 cttgaacagc tacggaactc ttgtgcgtaa 1470 <210> 148 <211> 489 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-cMyc Amino Acid Sequence <400> 148 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Leu Asp Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro 35 40 45 Ala Thr Met Pro Leu Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu 50 55 60 Asp Tyr Asp Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn 65 70 75 80 Phe Tyr Gln Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser 85 90 95 Glu Asp Ile Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser 100 105 110 Pro Ser Arg Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr 115 120 125 Pro Phe Ser Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser 130 135 140 Thr Ala Asp Gln Leu Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met 145 150 155 160 Val Asn Gln Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys 165 170 175 Asn Ile Ile Ile Gln Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala 180 185 190 Lys Leu Val Ser Glu Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp 195 200 205 Ser Gly Ser Pro Asn Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser 210 215 220 Ser Leu Tyr Leu Gln Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp 225 230 235 240 Pro Ser Val Val Phe Pro Tyr Pro Leu Asn Asp Ser Ser Ser Pro Lys 245 250 255 Ser Cys Ala Ser Gln Asp Ser Ser Ala Phe Ser Pro Ser Ser Asp Ser 260 265 270 Leu Leu Ser Ser Thr Glu Ser Ser Pro Gln Gly Ser Pro Glu Pro Leu 275 280 285 Val Leu His Glu Glu Thr Pro Pro Thr Thr Ser Ser Asp Ser Glu Glu 290 295 300 Glu Gln Glu Asp Glu Glu Glu Ile Asp Val Val Ser Val Glu Lys Arg 305 310 315 320 Gln Ala Pro Gly Lys Arg Ser Glu Ser Gly Ser Pro Ser Ala Gly Gly 325 330 335 His Ser Lys Pro Pro His Ser Pro Leu Val Leu Lys Arg Cys His Val 340 345 350 Ser Thr His Gln His Asn Tyr Ala Ala Pro Pro Ser Thr Arg Lys Asp 355 360 365 Tyr Pro Ala Ala Lys Arg Val Lys Leu Asp Ser Val Arg Val Leu Arg 370 375 380 Gln Ile Ser Asn Asn Arg Lys Cys Thr Ser Pro Arg Ser Ser Asp Thr 385 390 395 400 Glu Glu Asn Val Lys Arg Arg Thr His Asn Val Leu Glu Arg Gln Arg 405 410 415 Arg Asn Glu Leu Lys Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro 420 425 430 Glu Leu Glu Asn Asn Glu Lys Ala Pro Lys Val Val Ile Leu Lys Lys 435 440 445 Ala Thr Ala Tyr Ile Leu Ser Val Gln Ala Glu Glu Gln Lys Leu Ile 450 455 460 Ser Glu Glu Asp Leu Leu Arg Lys Arg Arg Glu Gln Leu Lys His Lys 465 470 475 480 Leu Glu Gln Leu Arg Asn Ser Cys Ala 485 <210> 149 <211> 1470 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-cMyc-JO-84 MTD cDNA Sequence <400> 149 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggattttttt cgggtagtgg aaaaccagca gcctcccgcg 120 acgatgcccc tcaacgttag cttcaccaac aggaactatg acctcgacta cgactcggtg 180 cagccgtatt tctactgcga cgaggaggag aacttctacc agcagcagca gcagagcgag 240 ctgcagcccc cggcgcccag cgaggatatc tggaagaaat tcgagctgct gcccaccccg 300 cccctgtccc ctagccgccg ctccgggctc tgctcgccct cctacgttgc ggtcacaccc 360 ttctcccttc ggggagacaa cgacggcggt ggcgggagct tctccacggc cgaccagctg 420 gagatggtga ccgagctgct gggaggagac atggtgaacc agagtttcat ctgcgacccg 480 gacgacgaga ccttcatcaa aaacatcatc atccaggact gtatgtggag cggcttctcg 540 gccgccgcca agctcgtctc agagaagctg gcctcctacc aggctgcgcg caaagacagc 600 ggcagcccga accccgcccg cggccacagc gtctgctcca cctccagctt gtacctgcag 660 gatctgagcg ccgccgcctc agagtgcatc gacccctcgg tggtcttccc ctaccctctc 720 aacgacagca gctcgcccaa gtcctgcgcc tcgcaagact ccagcgcctt ctctccgtcc 780 tcggattctc tgctctcctc gacggagtcc tccccgcagg gcagccccga gcccctggtg 840 ctccatgagg agacaccgcc caccaccagc agcgactctg aggaggaaca agaagatgag 900 gaagaaatcg atgttgtttc tgtggaaaag aggcaggctc ctggcaaaag gtcagagtct 960 ggatcacctt ctgctggagg ccacagcaaa cctcctcaca gcccactggt cctcaagagg 1020 tgccacgtct ccacacatca gcacaactac gcagcgcctc cctccactcg gaaggactat 1080 cctgctgcca agagggtcaa gttggacagt gtcagagtcc tgagacagat cagcaacaac 1140 cgaaaatgca ccagccccag gtcctcggac accgaggaga atgtcaagag gcgaacacac 1200 aacgtcttgg agcgccagag gaggaacgag ctaaaacgga gcttttttgc cctgcgtgac 1260 cagatcccgg agttggaaaa caatgaaaag gcccccaagg tagttatcct taaaaaagcc 1320 acagcataca tcctgtccgt ccaagcagag gagcaaaagc tcatttctga agaggacttg 1380 ttgcggaaac gacgagaaca gttgaaacac aaacttgaac agctacggaa ctcttgtgcg 1440 ctggtggcgg cgctgctggc ggtgctgtaa 1470 <210> 150 <211> 489 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-cMyc-JO-84 MTD Amino Acid Sequence <400> 150 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Asp Phe Phe Arg Val 20 25 30 Val Glu Asn Gln Gln Pro Pro Ala Thr Met Pro Leu Asn Val Ser Phe 35 40 45 Thr Asn Arg Asn Tyr Asp Leu Asp Tyr Asp Ser Val Gln Pro Tyr Phe 50 55 60 Tyr Cys Asp Glu Glu Glu Asn Phe Tyr Gln Gln Gln Gln Gln Ser Glu 65 70 75 80 Leu Gln Pro Pro Ala Pro Ser Glu Asp Ile Trp Lys Lys Phe Glu Leu 85 90 95 Leu Pro Thr Pro Pro Leu Ser Pro Ser Arg Arg Ser Gly Leu Cys Ser 100 105 110 Pro Ser Tyr Val Ala Val Thr Pro Phe Ser Leu Arg Gly Asp Asn Asp 115 120 125 Gly Gly Gly Gly Ser Phe Ser Thr Ala Asp Gln Leu Glu Met Val Thr 130 135 140 Glu Leu Leu Gly Gly Asp Met Val Asn Gln Ser Phe Ile Cys Asp Pro 145 150 155 160 Asp Asp Glu Thr Phe Ile Lys Asn Ile Ile Ile Gln Asp Cys Met Trp 165 170 175 Ser Gly Phe Ser Ala Ala Ala Lys Leu Val Ser Glu Lys Leu Ala Ser 180 185 190 Tyr Gln Ala Ala Arg Lys Asp Ser Gly Ser Pro Asn Pro Ala Arg Gly 195 200 205 His Ser Val Cys Ser Thr Ser Ser Leu Tyr Leu Gln Asp Leu Ser Ala 210 215 220 Ala Ala Ser Glu Cys Ile Asp Pro Ser Val Val Phe Pro Tyr Pro Leu 225 230 235 240 Asn Asp Ser Ser Ser Pro Lys Ser Cys Ala Ser Gln Asp Ser Ser Ala 245 250 255 Phe Ser Pro Ser Ser Asp Ser Leu Leu Ser Ser Thr Glu Ser Ser Pro 260 265 270 Gln Gly Ser Pro Glu Pro Leu Val Leu His Glu Glu Thr Pro Pro Thr 275 280 285 Thr Ser Ser Asp Ser Glu Glu Glu Gln Glu Asp Glu Glu Glu Ile Asp 290 295 300 Val Val Ser Val Glu Lys Arg Gln Ala Pro Gly Lys Arg Ser Glu Ser 305 310 315 320 Gly Ser Pro Ser Ala Gly Gly His Ser Lys Pro Pro His Ser Pro Leu 325 330 335 Val Leu Lys Arg Cys His Val Ser Thr His Gln His Asn Tyr Ala Ala 340 345 350 Pro Pro Ser Thr Arg Lys Asp Tyr Pro Ala Ala Lys Arg Val Lys Leu 355 360 365 Asp Ser Val Arg Val Leu Arg Gln Ile Ser Asn Asn Arg Lys Cys Thr 370 375 380 Ser Pro Arg Ser Ser Asp Thr Glu Glu Asn Val Lys Arg Arg Thr His 385 390 395 400 Asn Val Leu Glu Arg Gln Arg Arg Asn Glu Leu Lys Arg Ser Phe Phe 405 410 415 Ala Leu Arg Asp Gln Ile Pro Glu Leu Glu Asn Asn Glu Lys Ala Pro 420 425 430 Lys Val Val Ile Leu Lys Lys Ala Thr Ala Tyr Ile Leu Ser Val Gln 435 440 445 Ala Glu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Leu Arg Lys Arg 450 455 460 Arg Glu Gln Leu Lys His Lys Leu Glu Gln Leu Arg Asn Ser Cys Ala 465 470 475 480 Leu Val Ala Ala Leu Leu Ala Val Leu 485 <210> 151 <211> 1497 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-cMyc-JO-84 MTD cDNA Sequence <400> 151 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgctgga tttttttcgg 120 gtagtggaaa accagcagcc tcccgcgacg atgcccctca acgttagctt caccaacagg 180 aactatgacc tcgactacga ctcggtgcag ccgtatttct actgcgacga ggaggagaac 240 ttctaccagc agcagcagca gagcgagctg cagcccccgg cgcccagcga ggatatctgg 300 aagaaattcg agctgctgcc caccccgccc ctgtccccta gccgccgctc cgggctctgc 360 tcgccctcct acgttgcggt cacacccttc tcccttcggg gagacaacga cggcggtggc 420 gggagcttct ccacggccga ccagctggag atggtgaccg agctgctggg aggagacatg 480 gtgaaccaga gtttcatctg cgacccggac gacgagacct tcatcaaaaa catcatcatc 540 caggactgta tgtggagcgg cttctcggcc gccgccaagc tcgtctcaga gaagctggcc 600 tcctaccagg ctgcgcgcaa agacagcggc agcccgaacc ccgcccgcgg ccacagcgtc 660 tgctccacct ccagcttgta cctgcaggat ctgagcgccg ccgcctcaga gtgcatcgac 720 ccctcggtgg tcttccccta ccctctcaac gacagcagct cgcccaagtc ctgcgcctcg 780 caagactcca gcgccttctc tccgtcctcg gattctctgc tctcctcgac ggagtcctcc 840 ccgcagggca gccccgagcc cctggtgctc catgaggaga caccgcccac caccagcagc 900 gactctgagg aggaacaaga agatgaggaa gaaatcgatg ttgtttctgt ggaaaagagg 960 caggctcctg gcaaaaggtc agagtctgga tcaccttctg ctggaggcca cagcaaacct 1020 cctcacagcc cactggtcct caagaggtgc cacgtctcca cacatcagca caactacgca 1080 gcgcctccct ccactcggaa ggactatcct gctgccaaga gggtcaagtt ggacagtgtc 1140 agagtcctga gacagatcag caacaaccga aaatgcacca gccccaggtc ctcggacacc 1200 gaggagaatg tcaagaggcg aacacacaac gtcttggagc gccagaggag gaacgagcta 1260 aaacggagct tttttgccct gcgtgaccag atcccggagt tggaaaacaa tgaaaaggcc 1320 cccaaggtag ttatccttaa aaaagccaca gcatacatcc tgtccgtcca agcagaggag 1380 caaaagctca tttctgaaga ggacttgttg cggaaacgac gagaacagtt gaaacacaaa 1440 cttgaacagc tacggaactc ttgtgcgctg gtggcggcgc tgctggcggt gctgtaa 1497 <210> 152 <211> 498 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-cMyc-JO-84 MTD Amino Acid Sequence <400> 152 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Leu Asp Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro 35 40 45 Ala Thr Met Pro Leu Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu 50 55 60 Asp Tyr Asp Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn 65 70 75 80 Phe Tyr Gln Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser 85 90 95 Glu Asp Ile Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser 100 105 110 Pro Ser Arg Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr 115 120 125 Pro Phe Ser Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser 130 135 140 Thr Ala Asp Gln Leu Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met 145 150 155 160 Val Asn Gln Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys 165 170 175 Asn Ile Ile Ile Gln Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala 180 185 190 Lys Leu Val Ser Glu Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp 195 200 205 Ser Gly Ser Pro Asn Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser 210 215 220 Ser Leu Tyr Leu Gln Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp 225 230 235 240 Pro Ser Val Val Phe Pro Tyr Pro Leu Asn Asp Ser Ser Ser Pro Lys 245 250 255 Ser Cys Ala Ser Gln Asp Ser Ser Ala Phe Ser Pro Ser Ser Asp Ser 260 265 270 Leu Leu Ser Ser Thr Glu Ser Ser Pro Gln Gly Ser Pro Glu Pro Leu 275 280 285 Val Leu His Glu Glu Thr Pro Pro Thr Thr Ser Ser Asp Ser Glu Glu 290 295 300 Glu Gln Glu Asp Glu Glu Glu Ile Asp Val Val Ser Val Glu Lys Arg 305 310 315 320 Gln Ala Pro Gly Lys Arg Ser Glu Ser Gly Ser Pro Ser Ala Gly Gly 325 330 335 His Ser Lys Pro Pro His Ser Pro Leu Val Leu Lys Arg Cys His Val 340 345 350 Ser Thr His Gln His Asn Tyr Ala Ala Pro Pro Ser Thr Arg Lys Asp 355 360 365 Tyr Pro Ala Ala Lys Arg Val Lys Leu Asp Ser Val Arg Val Leu Arg 370 375 380 Gln Ile Ser Asn Asn Arg Lys Cys Thr Ser Pro Arg Ser Ser Asp Thr 385 390 395 400 Glu Glu Asn Val Lys Arg Arg Thr His Asn Val Leu Glu Arg Gln Arg 405 410 415 Arg Asn Glu Leu Lys Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro 420 425 430 Glu Leu Glu Asn Asn Glu Lys Ala Pro Lys Val Val Ile Leu Lys Lys 435 440 445 Ala Thr Ala Tyr Ile Leu Ser Val Gln Ala Glu Glu Gln Lys Leu Ile 450 455 460 Ser Glu Glu Asp Leu Leu Arg Lys Arg Arg Glu Gln Leu Lys His Lys 465 470 475 480 Leu Glu Gln Leu Arg Asn Ser Cys Ala Leu Val Ala Ala Leu Leu Ala 485 490 495 Val Leu <210> 153 <211> 1467 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-cMyc cDNA Sequence <400> 153 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgctggattt ttttcgggta 120 gtggaaaacc agcagcctcc cgcgacgatg cccctcaacg ttagcttcac caacaggaac 180 tatgacctcg actacgactc ggtgcagccg tatttctact gcgacgagga ggagaacttc 240 taccagcagc agcagcagag cgagctgcag cccccggcgc ccagcgagga tatctggaag 300 aaattcgagc tgctgcccac cccgcccctg tcccctagcc gccgctccgg gctctgctcg 360 ccctcctacg ttgcggtcac acccttctcc cttcggggag acaacgacgg cggtggcggg 420 agcttctcca cggccgacca gctggagatg gtgaccgagc tgctgggagg agacatggtg 480 aaccagagtt tcatctgcga cccggacgac gagaccttca tcaaaaacat catcatccag 540 gactgtatgt ggagcggctt ctcggccgcc gccaagctcg tctcagagaa gctggcctcc 600 taccaggctg cgcgcaaaga cagcggcagc ccgaaccccg cccgcggcca cagcgtctgc 660 tccacctcca gcttgtacct gcaggatctg agcgccgccg cctcagagtg catcgacccc 720 tcggtggtct tcccctaccc tctcaacgac agcagctcgc ccaagtcctg cgcctcgcaa 780 gactccagcg ccttctctcc gtcctcggat tctctgctct cctcgacgga gtcctccccg 840 cagggcagcc ccgagcccct ggtgctccat gaggagacac cgcccaccac cagcagcgac 900 tctgaggagg aacaagaaga tgaggaagaa atcgatgttg tttctgtgga aaagaggcag 960 gctcctggca aaaggtcaga gtctggatca ccttctgctg gaggccacag caaacctcct 1020 cacagcccac tggtcctcaa gaggtgccac gtctccacac atcagcacaa ctacgcagcg 1080 cctccctcca ctcggaagga ctatcctgct gccaagaggg tcaagttgga cagtgtcaga 1140 gtcctgagac agatcagcaa caaccgaaaa tgcaccagcc ccaggtcctc ggacaccgag 1200 gagaatgtca agaggcgaac acacaacgtc ttggagcgcc agaggaggaa cgagctaaaa 1260 cggagctttt ttgccctgcg tgaccagatc ccggagttgg aaaacaatga aaaggccccc 1320 aaggtagtta tccttaaaaa agccacagca tacatcctgt ccgtccaagc agaggagcaa 1380 aagctcattt ctgaagagga cttgttgcgg aaacgacgag aacagttgaa acacaaactt 1440 gaacagctac ggaactcttg tgcgtaa 1467 <210> 154 <211> 488 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-cMyc Amino Acid Sequence <400> 154 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Leu Asp Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro Ala 35 40 45 Thr Met Pro Leu Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu Asp 50 55 60 Tyr Asp Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn Phe 65 70 75 80 Tyr Gln Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser Glu 85 90 95 Asp Ile Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser Pro 100 105 110 Ser Arg Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr Pro 115 120 125 Phe Ser Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser Thr 130 135 140 Ala Asp Gln Leu Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met Val 145 150 155 160 Asn Gln Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys Asn 165 170 175 Ile Ile Ile Gln Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala Lys 180 185 190 Leu Val Ser Glu Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp Ser 195 200 205 Gly Ser Pro Asn Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser Ser 210 215 220 Leu Tyr Leu Gln Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp Pro 225 230 235 240 Ser Val Val Phe Pro Tyr Pro Leu Asn Asp Ser Ser Ser Pro Lys Ser 245 250 255 Cys Ala Ser Gln Asp Ser Ser Ala Phe Ser Pro Ser Ser Asp Ser Leu 260 265 270 Leu Ser Ser Thr Glu Ser Ser Pro Gln Gly Ser Pro Glu Pro Leu Val 275 280 285 Leu His Glu Glu Thr Pro Pro Thr Thr Ser Ser Asp Ser Glu Glu Glu 290 295 300 Gln Glu Asp Glu Glu Glu Ile Asp Val Val Ser Val Glu Lys Arg Gln 305 310 315 320 Ala Pro Gly Lys Arg Ser Glu Ser Gly Ser Pro Ser Ala Gly Gly His 325 330 335 Ser Lys Pro Pro His Ser Pro Leu Val Leu Lys Arg Cys His Val Ser 340 345 350 Thr His Gln His Asn Tyr Ala Ala Pro Pro Ser Thr Arg Lys Asp Tyr 355 360 365 Pro Ala Ala Lys Arg Val Lys Leu Asp Ser Val Arg Val Leu Arg Gln 370 375 380 Ile Ser Asn Asn Arg Lys Cys Thr Ser Pro Arg Ser Ser Asp Thr Glu 385 390 395 400 Glu Asn Val Lys Arg Arg Thr His Asn Val Leu Glu Arg Gln Arg Arg 405 410 415 Asn Glu Leu Lys Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro Glu 420 425 430 Leu Glu Asn Asn Glu Lys Ala Pro Lys Val Val Ile Leu Lys Lys Ala 435 440 445 Thr Ala Tyr Ile Leu Ser Val Gln Ala Glu Glu Gln Lys Leu Ile Ser 450 455 460 Glu Glu Asp Leu Leu Arg Lys Arg Arg Glu Gln Leu Lys His Lys Leu 465 470 475 480 Glu Gln Leu Arg Asn Ser Cys Ala 485 <210> 155 <211> 1467 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-cMyc-JO-86 MTD cDNA Sequence <400> 155 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggattttttt cgggtagtgg aaaaccagca gcctcccgcg 120 acgatgcccc tcaacgttag cttcaccaac aggaactatg acctcgacta cgactcggtg 180 cagccgtatt tctactgcga cgaggaggag aacttctacc agcagcagca gcagagcgag 240 ctgcagcccc cggcgcccag cgaggatatc tggaagaaat tcgagctgct gcccaccccg 300 cccctgtccc ctagccgccg ctccgggctc tgctcgccct cctacgttgc ggtcacaccc 360 ttctcccttc ggggagacaa cgacggcggt ggcgggagct tctccacggc cgaccagctg 420 gagatggtga ccgagctgct gggaggagac atggtgaacc agagtttcat ctgcgacccg 480 gacgacgaga ccttcatcaa aaacatcatc atccaggact gtatgtggag cggcttctcg 540 gccgccgcca agctcgtctc agagaagctg gcctcctacc aggctgcgcg caaagacagc 600 ggcagcccga accccgcccg cggccacagc gtctgctcca cctccagctt gtacctgcag 660 gatctgagcg ccgccgcctc agagtgcatc gacccctcgg tggtcttccc ctaccctctc 720 aacgacagca gctcgcccaa gtcctgcgcc tcgcaagact ccagcgcctt ctctccgtcc 780 tcggattctc tgctctcctc gacggagtcc tccccgcagg gcagccccga gcccctggtg 840 ctccatgagg agacaccgcc caccaccagc agcgactctg aggaggaaca agaagatgag 900 gaagaaatcg atgttgtttc tgtggaaaag aggcaggctc ctggcaaaag gtcagagtct 960 ggatcacctt ctgctggagg ccacagcaaa cctcctcaca gcccactggt cctcaagagg 1020 tgccacgtct ccacacatca gcacaactac gcagcgcctc cctccactcg gaaggactat 1080 cctgctgcca agagggtcaa gttggacagt gtcagagtcc tgagacagat cagcaacaac 1140 cgaaaatgca ccagccccag gtcctcggac accgaggaga atgtcaagag gcgaacacac 1200 aacgtcttgg agcgccagag gaggaacgag ctaaaacgga gcttttttgc cctgcgtgac 1260 cagatcccgg agttggaaaa caatgaaaag gcccccaagg tagttatcct taaaaaagcc 1320 acagcataca tcctgtccgt ccaagcagag gagcaaaagc tcatttctga agaggacttg 1380 ttgcggaaac gacgagaaca gttgaaacac aaacttgaac agctacggaa ctcttgtgcg 1440 ctggcggtgc tggcggcggc gccgtaa 1467 <210> 156 <211> 488 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-cMyc-JO-86 MTD Amino Acid Sequence <400> 156 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Asp Phe Phe Arg Val 20 25 30 Val Glu Asn Gln Gln Pro Pro Ala Thr Met Pro Leu Asn Val Ser Phe 35 40 45 Thr Asn Arg Asn Tyr Asp Leu Asp Tyr Asp Ser Val Gln Pro Tyr Phe 50 55 60 Tyr Cys Asp Glu Glu Glu Asn Phe Tyr Gln Gln Gln Gln Gln Ser Glu 65 70 75 80 Leu Gln Pro Pro Ala Pro Ser Glu Asp Ile Trp Lys Lys Phe Glu Leu 85 90 95 Leu Pro Thr Pro Pro Leu Ser Pro Ser Arg Arg Ser Gly Leu Cys Ser 100 105 110 Pro Ser Tyr Val Ala Val Thr Pro Phe Ser Leu Arg Gly Asp Asn Asp 115 120 125 Gly Gly Gly Gly Ser Phe Ser Thr Ala Asp Gln Leu Glu Met Val Thr 130 135 140 Glu Leu Leu Gly Gly Asp Met Val Asn Gln Ser Phe Ile Cys Asp Pro 145 150 155 160 Asp Asp Glu Thr Phe Ile Lys Asn Ile Ile Ile Gln Asp Cys Met Trp 165 170 175 Ser Gly Phe Ser Ala Ala Ala Lys Leu Val Ser Glu Lys Leu Ala Ser 180 185 190 Tyr Gln Ala Ala Arg Lys Asp Ser Gly Ser Pro Asn Pro Ala Arg Gly 195 200 205 His Ser Val Cys Ser Thr Ser Ser Leu Tyr Leu Gln Asp Leu Ser Ala 210 215 220 Ala Ala Ser Glu Cys Ile Asp Pro Ser Val Val Phe Pro Tyr Pro Leu 225 230 235 240 Asn Asp Ser Ser Ser Pro Lys Ser Cys Ala Ser Gln Asp Ser Ser Ala 245 250 255 Phe Ser Pro Ser Ser Asp Ser Leu Leu Ser Ser Thr Glu Ser Ser Pro 260 265 270 Gln Gly Ser Pro Glu Pro Leu Val Leu His Glu Glu Thr Pro Pro Thr 275 280 285 Thr Ser Ser Asp Ser Glu Glu Glu Gln Glu Asp Glu Glu Glu Ile Asp 290 295 300 Val Val Ser Val Glu Lys Arg Gln Ala Pro Gly Lys Arg Ser Glu Ser 305 310 315 320 Gly Ser Pro Ser Ala Gly Gly His Ser Lys Pro Pro His Ser Pro Leu 325 330 335 Val Leu Lys Arg Cys His Val Ser Thr His Gln His Asn Tyr Ala Ala 340 345 350 Pro Pro Ser Thr Arg Lys Asp Tyr Pro Ala Ala Lys Arg Val Lys Leu 355 360 365 Asp Ser Val Arg Val Leu Arg Gln Ile Ser Asn Asn Arg Lys Cys Thr 370 375 380 Ser Pro Arg Ser Ser Asp Thr Glu Glu Asn Val Lys Arg Arg Thr His 385 390 395 400 Asn Val Leu Glu Arg Gln Arg Arg Asn Glu Leu Lys Arg Ser Phe Phe 405 410 415 Ala Leu Arg Asp Gln Ile Pro Glu Leu Glu Asn Asn Glu Lys Ala Pro 420 425 430 Lys Val Val Ile Leu Lys Lys Ala Thr Ala Tyr Ile Leu Ser Val Gln 435 440 445 Ala Glu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Leu Arg Lys Arg 450 455 460 Arg Glu Gln Leu Lys His Lys Leu Glu Gln Leu Arg Asn Ser Cys Ala 465 470 475 480 Leu Ala Val Leu Ala Ala Ala Pro 485 <210> 157 <211> 1491 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-cMyc-JO-86 MTD cDNA Sequence <400> 157 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgctggattt ttttcgggta 120 gtggaaaacc agcagcctcc cgcgacgatg cccctcaacg ttagcttcac caacaggaac 180 tatgacctcg actacgactc ggtgcagccg tatttctact gcgacgagga ggagaacttc 240 taccagcagc agcagcagag cgagctgcag cccccggcgc ccagcgagga tatctggaag 300 aaattcgagc tgctgcccac cccgcccctg tcccctagcc gccgctccgg gctctgctcg 360 ccctcctacg ttgcggtcac acccttctcc cttcggggag acaacgacgg cggtggcggg 420 agcttctcca cggccgacca gctggagatg gtgaccgagc tgctgggagg agacatggtg 480 aaccagagtt tcatctgcga cccggacgac gagaccttca tcaaaaacat catcatccag 540 gactgtatgt ggagcggctt ctcggccgcc gccaagctcg tctcagagaa gctggcctcc 600 taccaggctg cgcgcaaaga cagcggcagc ccgaaccccg cccgcggcca cagcgtctgc 660 tccacctcca gcttgtacct gcaggatctg agcgccgccg cctcagagtg catcgacccc 720 tcggtggtct tcccctaccc tctcaacgac agcagctcgc ccaagtcctg cgcctcgcaa 780 gactccagcg ccttctctcc gtcctcggat tctctgctct cctcgacgga gtcctccccg 840 cagggcagcc ccgagcccct ggtgctccat gaggagacac cgcccaccac cagcagcgac 900 tctgaggagg aacaagaaga tgaggaagaa atcgatgttg tttctgtgga aaagaggcag 960 gctcctggca aaaggtcaga gtctggatca ccttctgctg gaggccacag caaacctcct 1020 cacagcccac tggtcctcaa gaggtgccac gtctccacac atcagcacaa ctacgcagcg 1080 cctccctcca ctcggaagga ctatcctgct gccaagaggg tcaagttgga cagtgtcaga 1140 gtcctgagac agatcagcaa caaccgaaaa tgcaccagcc ccaggtcctc ggacaccgag 1200 gagaatgtca agaggcgaac acacaacgtc ttggagcgcc agaggaggaa cgagctaaaa 1260 cggagctttt ttgccctgcg tgaccagatc ccggagttgg aaaacaatga aaaggccccc 1320 aaggtagtta tccttaaaaa agccacagca tacatcctgt ccgtccaagc agaggagcaa 1380 aagctcattt ctgaagagga cttgttgcgg aaacgacgag aacagttgaa acacaaactt 1440 gaacagctac ggaactcttg tgcgctggcg gtgctggcgg cggcgccgta a 1491 <210> 158 <211> 496 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-cMyc-JO-86 MTD Amino Acid Sequence <400> 158 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Leu Asp Phe Phe Arg Val Val Glu Asn Gln Gln Pro Pro Ala 35 40 45 Thr Met Pro Leu Asn Val Ser Phe Thr Asn Arg Asn Tyr Asp Leu Asp 50 55 60 Tyr Asp Ser Val Gln Pro Tyr Phe Tyr Cys Asp Glu Glu Glu Asn Phe 65 70 75 80 Tyr Gln Gln Gln Gln Gln Ser Glu Leu Gln Pro Pro Ala Pro Ser Glu 85 90 95 Asp Ile Trp Lys Lys Phe Glu Leu Leu Pro Thr Pro Pro Leu Ser Pro 100 105 110 Ser Arg Arg Ser Gly Leu Cys Ser Pro Ser Tyr Val Ala Val Thr Pro 115 120 125 Phe Ser Leu Arg Gly Asp Asn Asp Gly Gly Gly Gly Ser Phe Ser Thr 130 135 140 Ala Asp Gln Leu Glu Met Val Thr Glu Leu Leu Gly Gly Asp Met Val 145 150 155 160 Asn Gln Ser Phe Ile Cys Asp Pro Asp Asp Glu Thr Phe Ile Lys Asn 165 170 175 Ile Ile Ile Gln Asp Cys Met Trp Ser Gly Phe Ser Ala Ala Ala Lys 180 185 190 Leu Val Ser Glu Lys Leu Ala Ser Tyr Gln Ala Ala Arg Lys Asp Ser 195 200 205 Gly Ser Pro Asn Pro Ala Arg Gly His Ser Val Cys Ser Thr Ser Ser 210 215 220 Leu Tyr Leu Gln Asp Leu Ser Ala Ala Ala Ser Glu Cys Ile Asp Pro 225 230 235 240 Ser Val Val Phe Pro Tyr Pro Leu Asn Asp Ser Ser Ser Pro Lys Ser 245 250 255 Cys Ala Ser Gln Asp Ser Ser Ala Phe Ser Pro Ser Ser Asp Ser Leu 260 265 270 Leu Ser Ser Thr Glu Ser Ser Pro Gln Gly Ser Pro Glu Pro Leu Val 275 280 285 Leu His Glu Glu Thr Pro Pro Thr Thr Ser Ser Asp Ser Glu Glu Glu 290 295 300 Gln Glu Asp Glu Glu Glu Ile Asp Val Val Ser Val Glu Lys Arg Gln 305 310 315 320 Ala Pro Gly Lys Arg Ser Glu Ser Gly Ser Pro Ser Ala Gly Gly His 325 330 335 Ser Lys Pro Pro His Ser Pro Leu Val Leu Lys Arg Cys His Val Ser 340 345 350 Thr His Gln His Asn Tyr Ala Ala Pro Pro Ser Thr Arg Lys Asp Tyr 355 360 365 Pro Ala Ala Lys Arg Val Lys Leu Asp Ser Val Arg Val Leu Arg Gln 370 375 380 Ile Ser Asn Asn Arg Lys Cys Thr Ser Pro Arg Ser Ser Asp Thr Glu 385 390 395 400 Glu Asn Val Lys Arg Arg Thr His Asn Val Leu Glu Arg Gln Arg Arg 405 410 415 Asn Glu Leu Lys Arg Ser Phe Phe Ala Leu Arg Asp Gln Ile Pro Glu 420 425 430 Leu Glu Asn Asn Glu Lys Ala Pro Lys Val Val Ile Leu Lys Lys Ala 435 440 445 Thr Ala Tyr Ile Leu Ser Val Gln Ala Glu Glu Gln Lys Leu Ile Ser 450 455 460 Glu Glu Asp Leu Leu Arg Lys Arg Arg Glu Gln Leu Lys His Lys Leu 465 470 475 480 Glu Gln Leu Arg Asn Ser Cys Ala Leu Ala Val Leu Ala Ala Ala Pro 485 490 495 <210> 159 <211> 645 <212> DNA <213> Artificial Sequence <220> <223> NLS-Lin28 cDNA Sequence <400> 159 atgaagaaga agaggaaggg ctccgtgtcc aaccagcagt ttgcaggtgg ctgcgccaag 60 gcggcagaag aggcgcccga ggaggcgccg gaggacgcgg cccgggcggc ggacgagcct 120 cagctgctgc acggtgcggg catctgtaag tggttcaacg tgcgcatggg gttcggcttc 180 ctgtccatga ccgcccgcgc cggggtcgcg ctcgaccccc cagtggatgt ctttgtgcac 240 cagagtaagc tgcacatgga agggttccgg agcttgaagg agggtgaggc agtggagttc 300 acctttaaga agtcagccaa gggtctggaa tccatccgtg tcaccggacc tggtggagta 360 ttctgtattg ggagtgagag gcggccaaaa ggaaagagca tgcagaagcg cagatcaaaa 420 ggagacaggt gctacaactg tggaggtcta gatcatcatg ccaaggaatg caagctgcca 480 ccccagccca agaagtgcca cttctgccag agcatcagcc atatggtagc ctcatgtccg 540 ctgaaggccc agcagggccc tagtgcacag ggaaagccaa cctactttcg agaggaagaa 600 gaagaaatcc acagccctac cctgctcccg gaggcacaga attga 645 <210> 160 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> NLS-Lin28 Amino Acid Sequence <400> 160 Met Lys Lys Lys Arg Lys Gly Ser Val Ser Asn Gln Gln Phe Ala Gly 1 5 10 15 Gly Cys Ala Lys Ala Ala Glu Glu Ala Pro Glu Glu Ala Pro Glu Asp 20 25 30 Ala Ala Arg Ala Ala Asp Glu Pro Gln Leu Leu His Gly Ala Gly Ile 35 40 45 Cys Lys Trp Phe Asn Val Arg Met Gly Phe Gly Phe Leu Ser Met Thr 50 55 60 Ala Arg Ala Gly Val Ala Leu Asp Pro Pro Val Asp Val Phe Val His 65 70 75 80 Gln Ser Lys Leu His Met Glu Gly Phe Arg Ser Leu Lys Glu Gly Glu 85 90 95 Ala Val Glu Phe Thr Phe Lys Lys Ser Ala Lys Gly Leu Glu Ser Ile 100 105 110 Arg Val Thr Gly Pro Gly Gly Val Phe Cys Ile Gly Ser Glu Arg Arg 115 120 125 Pro Lys Gly Lys Ser Met Gln Lys Arg Arg Ser Lys Gly Asp Arg Cys 130 135 140 Tyr Asn Cys Gly Gly Leu Asp His His Ala Lys Glu Cys Lys Leu Pro 145 150 155 160 Pro Gln Pro Lys Lys Cys His Phe Cys Gln Ser Ile Ser His Met Val 165 170 175 Ala Ser Cys Pro Leu Lys Ala Gln Gln Gly Pro Ser Ala Gln Gly Lys 180 185 190 Pro Thr Tyr Phe Arg Glu Glu Glu Glu Glu Ile His Ser Pro Thr Leu 195 200 205 Leu Pro Glu Ala Gln Asn 210 <210> 161 <211> 672 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Lin28 cDNA Sequence <400> 161 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgggctc cgtgtccaac 60 cagcagtttg caggtggctg cgccaaggcg gcagaagagg cgcccgagga ggcgccggag 120 gacgcggccc gggcggcgga cgagcctcag ctgctgcacg gtgcgggcat ctgtaagtgg 180 ttcaacgtgc gcatggggtt cggcttcctg tccatgaccg cccgcgccgg ggtcgcgctc 240 gaccccccag tggatgtctt tgtgcaccag agtaagctgc acatggaagg gttccggagc 300 ttgaaggagg gtgaggcagt ggagttcacc tttaagaagt cagccaaggg tctggaatcc 360 atccgtgtca ccggacctgg tggagtattc tgtattggga gtgagaggcg gccaaaagga 420 aagagcatgc agaagcgcag atcaaaagga gacaggtgct acaactgtgg aggtctagat 480 catcatgcca aggaatgcaa gctgccaccc cagcccaaga agtgccactt ctgccagagc 540 atcagccata tggtagcctc atgtccgctg aaggcccagc agggccctag tgcacaggga 600 aagccaacct actttcgaga ggaagaagaa gaaatccaca gccctaccct gctcccggag 660 gcacagaatt ga 672 <210> 162 <211> 223 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Lin28 Amino Acid Sequence <400> 162 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Gly 1 5 10 15 Ser Val Ser Asn Gln Gln Phe Ala Gly Gly Cys Ala Lys Ala Ala Glu 20 25 30 Glu Ala Pro Glu Glu Ala Pro Glu Asp Ala Ala Arg Ala Ala Asp Glu 35 40 45 Pro Gln Leu Leu His Gly Ala Gly Ile Cys Lys Trp Phe Asn Val Arg 50 55 60 Met Gly Phe Gly Phe Leu Ser Met Thr Ala Arg Ala Gly Val Ala Leu 65 70 75 80 Asp Pro Pro Val Asp Val Phe Val His Gln Ser Lys Leu His Met Glu 85 90 95 Gly Phe Arg Ser Leu Lys Glu Gly Glu Ala Val Glu Phe Thr Phe Lys 100 105 110 Lys Ser Ala Lys Gly Leu Glu Ser Ile Arg Val Thr Gly Pro Gly Gly 115 120 125 Val Phe Cys Ile Gly Ser Glu Arg Arg Pro Lys Gly Lys Ser Met Gln 130 135 140 Lys Arg Arg Ser Lys Gly Asp Arg Cys Tyr Asn Cys Gly Gly Leu Asp 145 150 155 160 His His Ala Lys Glu Cys Lys Leu Pro Pro Gln Pro Lys Lys Cys His 165 170 175 Phe Cys Gln Ser Ile Ser His Met Val Ala Ser Cys Pro Leu Lys Ala 180 185 190 Gln Gln Gly Pro Ser Ala Gln Gly Lys Pro Thr Tyr Phe Arg Glu Glu 195 200 205 Glu Glu Glu Ile His Ser Pro Thr Leu Leu Pro Glu Ala Gln Asn 210 215 220 <210> 163 <211> 672 <212> DNA <213> Artificial Sequence <220> <223> NLS-Lin28-JO-84 MTD cDNA Sequence <400> 163 atgaagaaga agaggaaggg ctccgtgtcc aaccagcagt ttgcaggtgg ctgcgccaag 60 gcggcagaag aggcgcccga ggaggcgccg gaggacgcgg cccgggcggc ggacgagcct 120 cagctgctgc acggtgcggg catctgtaag tggttcaacg tgcgcatggg gttcggcttc 180 ctgtccatga ccgcccgcgc cggggtcgcg ctcgaccccc cagtggatgt ctttgtgcac 240 cagagtaagc tgcacatgga agggttccgg agcttgaagg agggtgaggc agtggagttc 300 acctttaaga agtcagccaa gggtctggaa tccatccgtg tcaccggacc tggtggagta 360 ttctgtattg ggagtgagag gcggccaaaa ggaaagagca tgcagaagcg cagatcaaaa 420 ggagacaggt gctacaactg tggaggtcta gatcatcatg ccaaggaatg caagctgcca 480 ccccagccca agaagtgcca cttctgccag agcatcagcc atatggtagc ctcatgtccg 540 ctgaaggccc agcagggccc tagtgcacag ggaaagccaa cctactttcg agaggaagaa 600 gaagaaatcc acagccctac cctgctcccg gaggcacaga atctggtggc ggcgctgctg 660 gcggtgctgt ga 672 <210> 164 <211> 223 <212> PRT <213> Artificial Sequence <220> <223> NLS-Lin28-JO-84 MTD Amino Acid Sequence <400> 164 Met Lys Lys Lys Arg Lys Gly Ser Val Ser Asn Gln Gln Phe Ala Gly 1 5 10 15 Gly Cys Ala Lys Ala Ala Glu Glu Ala Pro Glu Glu Ala Pro Glu Asp 20 25 30 Ala Ala Arg Ala Ala Asp Glu Pro Gln Leu Leu His Gly Ala Gly Ile 35 40 45 Cys Lys Trp Phe Asn Val Arg Met Gly Phe Gly Phe Leu Ser Met Thr 50 55 60 Ala Arg Ala Gly Val Ala Leu Asp Pro Pro Val Asp Val Phe Val His 65 70 75 80 Gln Ser Lys Leu His Met Glu Gly Phe Arg Ser Leu Lys Glu Gly Glu 85 90 95 Ala Val Glu Phe Thr Phe Lys Lys Ser Ala Lys Gly Leu Glu Ser Ile 100 105 110 Arg Val Thr Gly Pro Gly Gly Val Phe Cys Ile Gly Ser Glu Arg Arg 115 120 125 Pro Lys Gly Lys Ser Met Gln Lys Arg Arg Ser Lys Gly Asp Arg Cys 130 135 140 Tyr Asn Cys Gly Gly Leu Asp His His Ala Lys Glu Cys Lys Leu Pro 145 150 155 160 Pro Gln Pro Lys Lys Cys His Phe Cys Gln Ser Ile Ser His Met Val 165 170 175 Ala Ser Cys Pro Leu Lys Ala Gln Gln Gly Pro Ser Ala Gln Gly Lys 180 185 190 Pro Thr Tyr Phe Arg Glu Glu Glu Glu Glu Ile His Ser Pro Thr Leu 195 200 205 Leu Pro Glu Ala Gln Asn Leu Val Ala Ala Leu Leu Ala Val Leu 210 215 220 <210> 165 <211> 699 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Lin28-JO-84 MTD cDNA Sequence <400> 165 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgggctc cgtgtccaac 60 cagcagtttg caggtggctg cgccaaggcg gcagaagagg cgcccgagga ggcgccggag 120 gacgcggccc gggcggcgga cgagcctcag ctgctgcacg gtgcgggcat ctgtaagtgg 180 ttcaacgtgc gcatggggtt cggcttcctg tccatgaccg cccgcgccgg ggtcgcgctc 240 gaccccccag tggatgtctt tgtgcaccag agtaagctgc acatggaagg gttccggagc 300 ttgaaggagg gtgaggcagt ggagttcacc tttaagaagt cagccaaggg tctggaatcc 360 atccgtgtca ccggacctgg tggagtattc tgtattggga gtgagaggcg gccaaaagga 420 aagagcatgc agaagcgcag atcaaaagga gacaggtgct acaactgtgg aggtctagat 480 catcatgcca aggaatgcaa gctgccaccc cagcccaaga agtgccactt ctgccagagc 540 atcagccata tggtagcctc atgtccgctg aaggcccagc agggccctag tgcacaggga 600 aagccaacct actttcgaga ggaagaagaa gaaatccaca gccctaccct gctcccggag 660 gcacagaatc tggtggcggc gctgctggcg gtgctgtga 699 <210> 166 <211> 232 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-84 MTD-Lin28-JO-84 MTD Amino Acid Sequence <400> 166 Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu Ala Val Leu Gly 1 5 10 15 Ser Val Ser Asn Gln Gln Phe Ala Gly Gly Cys Ala Lys Ala Ala Glu 20 25 30 Glu Ala Pro Glu Glu Ala Pro Glu Asp Ala Ala Arg Ala Ala Asp Glu 35 40 45 Pro Gln Leu Leu His Gly Ala Gly Ile Cys Lys Trp Phe Asn Val Arg 50 55 60 Met Gly Phe Gly Phe Leu Ser Met Thr Ala Arg Ala Gly Val Ala Leu 65 70 75 80 Asp Pro Pro Val Asp Val Phe Val His Gln Ser Lys Leu His Met Glu 85 90 95 Gly Phe Arg Ser Leu Lys Glu Gly Glu Ala Val Glu Phe Thr Phe Lys 100 105 110 Lys Ser Ala Lys Gly Leu Glu Ser Ile Arg Val Thr Gly Pro Gly Gly 115 120 125 Val Phe Cys Ile Gly Ser Glu Arg Arg Pro Lys Gly Lys Ser Met Gln 130 135 140 Lys Arg Arg Ser Lys Gly Asp Arg Cys Tyr Asn Cys Gly Gly Leu Asp 145 150 155 160 His His Ala Lys Glu Cys Lys Leu Pro Pro Gln Pro Lys Lys Cys His 165 170 175 Phe Cys Gln Ser Ile Ser His Met Val Ala Ser Cys Pro Leu Lys Ala 180 185 190 Gln Gln Gly Pro Ser Ala Gln Gly Lys Pro Thr Tyr Phe Arg Glu Glu 195 200 205 Glu Glu Glu Ile His Ser Pro Thr Leu Leu Pro Glu Ala Gln Asn Leu 210 215 220 Val Ala Ala Leu Leu Ala Val Leu 225 230 <210> 167 <211> 669 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Lin28 cDNA Sequence <400> 167 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgggctccgt gtccaaccag 60 cagtttgcag gtggctgcgc caaggcggca gaagaggcgc ccgaggaggc gccggaggac 120 gcggcccggg cggcggacga gcctcagctg ctgcacggtg cgggcatctg taagtggttc 180 aacgtgcgca tggggttcgg cttcctgtcc atgaccgccc gcgccggggt cgcgctcgac 240 cccccagtgg atgtctttgt gcaccagagt aagctgcaca tggaagggtt ccggagcttg 300 aaggagggtg aggcagtgga gttcaccttt aagaagtcag ccaagggtct ggaatccatc 360 cgtgtcaccg gacctggtgg agtattctgt attgggagtg agaggcggcc aaaaggaaag 420 agcatgcaga agcgcagatc aaaaggagac aggtgctaca actgtggagg tctagatcat 480 catgccaagg aatgcaagct gccaccccag cccaagaagt gccacttctg ccagagcatc 540 agccatatgg tagcctcatg tccgctgaag gcccagcagg gccctagtgc acagggaaag 600 ccaacctact ttcgagagga agaagaagaa atccacagcc ctaccctgct cccggaggca 660 cagaattga 669 <210> 168 <211> 222 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Lin28 Amino Acid Sequence <400> 168 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Gly Ser 1 5 10 15 Val Ser Asn Gln Gln Phe Ala Gly Gly Cys Ala Lys Ala Ala Glu Glu 20 25 30 Ala Pro Glu Glu Ala Pro Glu Asp Ala Ala Arg Ala Ala Asp Glu Pro 35 40 45 Gln Leu Leu His Gly Ala Gly Ile Cys Lys Trp Phe Asn Val Arg Met 50 55 60 Gly Phe Gly Phe Leu Ser Met Thr Ala Arg Ala Gly Val Ala Leu Asp 65 70 75 80 Pro Pro Val Asp Val Phe Val His Gln Ser Lys Leu His Met Glu Gly 85 90 95 Phe Arg Ser Leu Lys Glu Gly Glu Ala Val Glu Phe Thr Phe Lys Lys 100 105 110 Ser Ala Lys Gly Leu Glu Ser Ile Arg Val Thr Gly Pro Gly Gly Val 115 120 125 Phe Cys Ile Gly Ser Glu Arg Arg Pro Lys Gly Lys Ser Met Gln Lys 130 135 140 Arg Arg Ser Lys Gly Asp Arg Cys Tyr Asn Cys Gly Gly Leu Asp His 145 150 155 160 His Ala Lys Glu Cys Lys Leu Pro Pro Gln Pro Lys Lys Cys His Phe 165 170 175 Cys Gln Ser Ile Ser His Met Val Ala Ser Cys Pro Leu Lys Ala Gln 180 185 190 Gln Gly Pro Ser Ala Gln Gly Lys Pro Thr Tyr Phe Arg Glu Glu Glu 195 200 205 Glu Glu Ile His Ser Pro Thr Leu Leu Pro Glu Ala Gln Asn 210 215 220 <210> 169 <211> 669 <212> DNA <213> Artificial Sequence <220> <223> NLS-Lin28-JO-86 MTD cDNA Sequence <400> 169 atgaagaaga agaggaaggg ctccgtgtcc aaccagcagt ttgcaggtgg ctgcgccaag 60 gcggcagaag aggcgcccga ggaggcgccg gaggacgcgg cccgggcggc ggacgagcct 120 cagctgctgc acggtgcggg catctgtaag tggttcaacg tgcgcatggg gttcggcttc 180 ctgtccatga ccgcccgcgc cggggtcgcg ctcgaccccc cagtggatgt ctttgtgcac 240 cagagtaagc tgcacatgga agggttccgg agcttgaagg agggtgaggc agtggagttc 300 acctttaaga agtcagccaa gggtctggaa tccatccgtg tcaccggacc tggtggagta 360 ttctgtattg ggagtgagag gcggccaaaa ggaaagagca tgcagaagcg cagatcaaaa 420 ggagacaggt gctacaactg tggaggtcta gatcatcatg ccaaggaatg caagctgcca 480 ccccagccca agaagtgcca cttctgccag agcatcagcc atatggtagc ctcatgtccg 540 ctgaaggccc agcagggccc tagtgcacag ggaaagccaa cctactttcg agaggaagaa 600 gaagaaatcc acagccctac cctgctcccg gaggcacaga atctggcggt gctggcggcg 660 gcgccgtga 669 <210> 170 <211> 222 <212> PRT <213> Artificial Sequence <220> <223> NLS-Lin28-JO-86 MTD Amino Acid Sequence <400> 170 Met Lys Lys Lys Arg Lys Gly Ser Val Ser Asn Gln Gln Phe Ala Gly 1 5 10 15 Gly Cys Ala Lys Ala Ala Glu Glu Ala Pro Glu Glu Ala Pro Glu Asp 20 25 30 Ala Ala Arg Ala Ala Asp Glu Pro Gln Leu Leu His Gly Ala Gly Ile 35 40 45 Cys Lys Trp Phe Asn Val Arg Met Gly Phe Gly Phe Leu Ser Met Thr 50 55 60 Ala Arg Ala Gly Val Ala Leu Asp Pro Pro Val Asp Val Phe Val His 65 70 75 80 Gln Ser Lys Leu His Met Glu Gly Phe Arg Ser Leu Lys Glu Gly Glu 85 90 95 Ala Val Glu Phe Thr Phe Lys Lys Ser Ala Lys Gly Leu Glu Ser Ile 100 105 110 Arg Val Thr Gly Pro Gly Gly Val Phe Cys Ile Gly Ser Glu Arg Arg 115 120 125 Pro Lys Gly Lys Ser Met Gln Lys Arg Arg Ser Lys Gly Asp Arg Cys 130 135 140 Tyr Asn Cys Gly Gly Leu Asp His His Ala Lys Glu Cys Lys Leu Pro 145 150 155 160 Pro Gln Pro Lys Lys Cys His Phe Cys Gln Ser Ile Ser His Met Val 165 170 175 Ala Ser Cys Pro Leu Lys Ala Gln Gln Gly Pro Ser Ala Gln Gly Lys 180 185 190 Pro Thr Tyr Phe Arg Glu Glu Glu Glu Glu Ile His Ser Pro Thr Leu 195 200 205 Leu Pro Glu Ala Gln Asn Leu Ala Val Leu Ala Ala Ala Pro 210 215 220 <210> 171 <211> 693 <212> DNA <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Lin28-JO-86 MTD cDNA Sequence <400> 171 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgggctccgt gtccaaccag 60 cagtttgcag gtggctgcgc caaggcggca gaagaggcgc ccgaggaggc gccggaggac 120 gcggcccggg cggcggacga gcctcagctg ctgcacggtg cgggcatctg taagtggttc 180 aacgtgcgca tggggttcgg cttcctgtcc atgaccgccc gcgccggggt cgcgctcgac 240 cccccagtgg atgtctttgt gcaccagagt aagctgcaca tggaagggtt ccggagcttg 300 aaggagggtg aggcagtgga gttcaccttt aagaagtcag ccaagggtct ggaatccatc 360 cgtgtcaccg gacctggtgg agtattctgt attgggagtg agaggcggcc aaaaggaaag 420 agcatgcaga agcgcagatc aaaaggagac aggtgctaca actgtggagg tctagatcat 480 catgccaagg aatgcaagct gccaccccag cccaagaagt gccacttctg ccagagcatc 540 agccatatgg tagcctcatg tccgctgaag gcccagcagg gccctagtgc acagggaaag 600 ccaacctact ttcgagagga agaagaagaa atccacagcc ctaccctgct cccggaggca 660 cagaatctgg cggtgctggc ggcggcgccg tga 693 <210> 172 <211> 230 <212> PRT <213> Artificial Sequence <220> <223> NLS-JO-86 MTD-Lin28-JO-86 MTD Amino Acid Sequence <400> 172 Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala Ala Pro Gly Ser 1 5 10 15 Val Ser Asn Gln Gln Phe Ala Gly Gly Cys Ala Lys Ala Ala Glu Glu 20 25 30 Ala Pro Glu Glu Ala Pro Glu Asp Ala Ala Arg Ala Ala Asp Glu Pro 35 40 45 Gln Leu Leu His Gly Ala Gly Ile Cys Lys Trp Phe Asn Val Arg Met 50 55 60 Gly Phe Gly Phe Leu Ser Met Thr Ala Arg Ala Gly Val Ala Leu Asp 65 70 75 80 Pro Pro Val Asp Val Phe Val His Gln Ser Lys Leu His Met Glu Gly 85 90 95 Phe Arg Ser Leu Lys Glu Gly Glu Ala Val Glu Phe Thr Phe Lys Lys 100 105 110 Ser Ala Lys Gly Leu Glu Ser Ile Arg Val Thr Gly Pro Gly Gly Val 115 120 125 Phe Cys Ile Gly Ser Glu Arg Arg Pro Lys Gly Lys Ser Met Gln Lys 130 135 140 Arg Arg Ser Lys Gly Asp Arg Cys Tyr Asn Cys Gly Gly Leu Asp His 145 150 155 160 His Ala Lys Glu Cys Lys Leu Pro Pro Gln Pro Lys Lys Cys His Phe 165 170 175 Cys Gln Ser Ile Ser His Met Val Ala Ser Cys Pro Leu Lys Ala Gln 180 185 190 Gln Gly Pro Ser Ala Gln Gly Lys Pro Thr Tyr Phe Arg Glu Glu Glu 195 200 205 Glu Glu Ile His Ser Pro Thr Leu Leu Pro Glu Ala Gln Asn Leu Ala 210 215 220 Val Leu Ala Ala Ala Pro 225 230 <210> 173 <211> 705 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Lin28 cDNA Sequence <400> 173 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaaggg ctccgtgtcc aaccagcagt ttgcaggtgg ctgcgccaag 120 gcggcagaag aggcgcccga ggaggcgccg gaggacgcgg cccgggcggc ggacgagcct 180 cagctgctgc acggtgcggg catctgtaag tggttcaacg tgcgcatggg gttcggcttc 240 ctgtccatga ccgcccgcgc cggggtcgcg ctcgaccccc cagtggatgt ctttgtgcac 300 cagagtaagc tgcacatgga agggttccgg agcttgaagg agggtgaggc agtggagttc 360 acctttaaga agtcagccaa gggtctggaa tccatccgtg tcaccggacc tggtggagta 420 ttctgtattg ggagtgagag gcggccaaaa ggaaagagca tgcagaagcg cagatcaaaa 480 ggagacaggt gctacaactg tggaggtcta gatcatcatg ccaaggaatg caagctgcca 540 ccccagccca agaagtgcca cttctgccag agcatcagcc atatggtagc ctcatgtccg 600 ctgaaggccc agcagggccc tagtgcacag ggaaagccaa cctactttcg agaggaagaa 660 gaagaaatcc acagccctac cctgctcccg gaggcacaga attga 705 <210> 174 <211> 234 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Lin28 Amino Acid Sequence <400> 174 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Gly Ser Val Ser Asn Gln 20 25 30 Gln Phe Ala Gly Gly Cys Ala Lys Ala Ala Glu Glu Ala Pro Glu Glu 35 40 45 Ala Pro Glu Asp Ala Ala Arg Ala Ala Asp Glu Pro Gln Leu Leu His 50 55 60 Gly Ala Gly Ile Cys Lys Trp Phe Asn Val Arg Met Gly Phe Gly Phe 65 70 75 80 Leu Ser Met Thr Ala Arg Ala Gly Val Ala Leu Asp Pro Pro Val Asp 85 90 95 Val Phe Val His Gln Ser Lys Leu His Met Glu Gly Phe Arg Ser Leu 100 105 110 Lys Glu Gly Glu Ala Val Glu Phe Thr Phe Lys Lys Ser Ala Lys Gly 115 120 125 Leu Glu Ser Ile Arg Val Thr Gly Pro Gly Gly Val Phe Cys Ile Gly 130 135 140 Ser Glu Arg Arg Pro Lys Gly Lys Ser Met Gln Lys Arg Arg Ser Lys 145 150 155 160 Gly Asp Arg Cys Tyr Asn Cys Gly Gly Leu Asp His His Ala Lys Glu 165 170 175 Cys Lys Leu Pro Pro Gln Pro Lys Lys Cys His Phe Cys Gln Ser Ile 180 185 190 Ser His Met Val Ala Ser Cys Pro Leu Lys Ala Gln Gln Gly Pro Ser 195 200 205 Ala Gln Gly Lys Pro Thr Tyr Phe Arg Glu Glu Glu Glu Glu Ile His 210 215 220 Ser Pro Thr Leu Leu Pro Glu Ala Gln Asn 225 230 <210> 175 <211> 732 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Lin28 cDNA Sequence <400> 175 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgggctc cgtgtccaac 120 cagcagtttg caggtggctg cgccaaggcg gcagaagagg cgcccgagga ggcgccggag 180 gacgcggccc gggcggcgga cgagcctcag ctgctgcacg gtgcgggcat ctgtaagtgg 240 ttcaacgtgc gcatggggtt cggcttcctg tccatgaccg cccgcgccgg ggtcgcgctc 300 gaccccccag tggatgtctt tgtgcaccag agtaagctgc acatggaagg gttccggagc 360 ttgaaggagg gtgaggcagt ggagttcacc tttaagaagt cagccaaggg tctggaatcc 420 atccgtgtca ccggacctgg tggagtattc tgtattggga gtgagaggcg gccaaaagga 480 aagagcatgc agaagcgcag atcaaaagga gacaggtgct acaactgtgg aggtctagat 540 catcatgcca aggaatgcaa gctgccaccc cagcccaaga agtgccactt ctgccagagc 600 atcagccata tggtagcctc atgtccgctg aaggcccagc agggccctag tgcacaggga 660 aagccaacct actttcgaga ggaagaagaa gaaatccaca gccctaccct gctcccggag 720 gcacagaatt ga 732 <210> 176 <211> 243 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Lin28 Amino Acid Sequence <400> 176 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Gly Ser Val Ser Asn Gln Gln Phe Ala Gly Gly Cys Ala 35 40 45 Lys Ala Ala Glu Glu Ala Pro Glu Glu Ala Pro Glu Asp Ala Ala Arg 50 55 60 Ala Ala Asp Glu Pro Gln Leu Leu His Gly Ala Gly Ile Cys Lys Trp 65 70 75 80 Phe Asn Val Arg Met Gly Phe Gly Phe Leu Ser Met Thr Ala Arg Ala 85 90 95 Gly Val Ala Leu Asp Pro Pro Val Asp Val Phe Val His Gln Ser Lys 100 105 110 Leu His Met Glu Gly Phe Arg Ser Leu Lys Glu Gly Glu Ala Val Glu 115 120 125 Phe Thr Phe Lys Lys Ser Ala Lys Gly Leu Glu Ser Ile Arg Val Thr 130 135 140 Gly Pro Gly Gly Val Phe Cys Ile Gly Ser Glu Arg Arg Pro Lys Gly 145 150 155 160 Lys Ser Met Gln Lys Arg Arg Ser Lys Gly Asp Arg Cys Tyr Asn Cys 165 170 175 Gly Gly Leu Asp His His Ala Lys Glu Cys Lys Leu Pro Pro Gln Pro 180 185 190 Lys Lys Cys His Phe Cys Gln Ser Ile Ser His Met Val Ala Ser Cys 195 200 205 Pro Leu Lys Ala Gln Gln Gly Pro Ser Ala Gln Gly Lys Pro Thr Tyr 210 215 220 Phe Arg Glu Glu Glu Glu Glu Ile His Ser Pro Thr Leu Leu Pro Glu 225 230 235 240 Ala Gln Asn <210> 177 <211> 732 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Lin28-JO-84 MTD cDNA Sequence <400> 177 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaaggg ctccgtgtcc aaccagcagt ttgcaggtgg ctgcgccaag 120 gcggcagaag aggcgcccga ggaggcgccg gaggacgcgg cccgggcggc ggacgagcct 180 cagctgctgc acggtgcggg catctgtaag tggttcaacg tgcgcatggg gttcggcttc 240 ctgtccatga ccgcccgcgc cggggtcgcg ctcgaccccc cagtggatgt ctttgtgcac 300 cagagtaagc tgcacatgga agggttccgg agcttgaagg agggtgaggc agtggagttc 360 acctttaaga agtcagccaa gggtctggaa tccatccgtg tcaccggacc tggtggagta 420 ttctgtattg ggagtgagag gcggccaaaa ggaaagagca tgcagaagcg cagatcaaaa 480 ggagacaggt gctacaactg tggaggtcta gatcatcatg ccaaggaatg caagctgcca 540 ccccagccca agaagtgcca cttctgccag agcatcagcc atatggtagc ctcatgtccg 600 ctgaaggccc agcagggccc tagtgcacag ggaaagccaa cctactttcg agaggaagaa 660 gaagaaatcc acagccctac cctgctcccg gaggcacaga atctggtggc ggcgctgctg 720 gcggtgctgt ga 732 <210> 178 <211> 243 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Lin28-JO-84 MTD Amino Acid Sequence <400> 178 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Gly Ser Val Ser Asn Gln 20 25 30 Gln Phe Ala Gly Gly Cys Ala Lys Ala Ala Glu Glu Ala Pro Glu Glu 35 40 45 Ala Pro Glu Asp Ala Ala Arg Ala Ala Asp Glu Pro Gln Leu Leu His 50 55 60 Gly Ala Gly Ile Cys Lys Trp Phe Asn Val Arg Met Gly Phe Gly Phe 65 70 75 80 Leu Ser Met Thr Ala Arg Ala Gly Val Ala Leu Asp Pro Pro Val Asp 85 90 95 Val Phe Val His Gln Ser Lys Leu His Met Glu Gly Phe Arg Ser Leu 100 105 110 Lys Glu Gly Glu Ala Val Glu Phe Thr Phe Lys Lys Ser Ala Lys Gly 115 120 125 Leu Glu Ser Ile Arg Val Thr Gly Pro Gly Gly Val Phe Cys Ile Gly 130 135 140 Ser Glu Arg Arg Pro Lys Gly Lys Ser Met Gln Lys Arg Arg Ser Lys 145 150 155 160 Gly Asp Arg Cys Tyr Asn Cys Gly Gly Leu Asp His His Ala Lys Glu 165 170 175 Cys Lys Leu Pro Pro Gln Pro Lys Lys Cys His Phe Cys Gln Ser Ile 180 185 190 Ser His Met Val Ala Ser Cys Pro Leu Lys Ala Gln Gln Gly Pro Ser 195 200 205 Ala Gln Gly Lys Pro Thr Tyr Phe Arg Glu Glu Glu Glu Glu Ile His 210 215 220 Ser Pro Thr Leu Leu Pro Glu Ala Gln Asn Leu Val Ala Ala Leu Leu 225 230 235 240 Ala Val Leu <210> 179 <211> 759 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Lin28-JO-84 MTD cDNA Sequence <400> 179 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggtggcggcg ctgctggcgg tgctgggctc cgtgtccaac 120 cagcagtttg caggtggctg cgccaaggcg gcagaagagg cgcccgagga ggcgccggag 180 gacgcggccc gggcggcgga cgagcctcag ctgctgcacg gtgcgggcat ctgtaagtgg 240 ttcaacgtgc gcatggggtt cggcttcctg tccatgaccg cccgcgccgg ggtcgcgctc 300 gaccccccag tggatgtctt tgtgcaccag agtaagctgc acatggaagg gttccggagc 360 ttgaaggagg gtgaggcagt ggagttcacc tttaagaagt cagccaaggg tctggaatcc 420 atccgtgtca ccggacctgg tggagtattc tgtattggga gtgagaggcg gccaaaagga 480 aagagcatgc agaagcgcag atcaaaagga gacaggtgct acaactgtgg aggtctagat 540 catcatgcca aggaatgcaa gctgccaccc cagcccaaga agtgccactt ctgccagagc 600 atcagccata tggtagcctc atgtccgctg aaggcccagc agggccctag tgcacaggga 660 aagccaacct actttcgaga ggaagaagaa gaaatccaca gccctaccct gctcccggag 720 gcacagaatc tggtggcggc gctgctggcg gtgctgtga 759 <210> 180 <211> 252 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-84 MTD-Lin28-JO-84 MTD Amino Acid Sequence <400> 180 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Val Ala Ala Leu Leu 20 25 30 Ala Val Leu Gly Ser Val Ser Asn Gln Gln Phe Ala Gly Gly Cys Ala 35 40 45 Lys Ala Ala Glu Glu Ala Pro Glu Glu Ala Pro Glu Asp Ala Ala Arg 50 55 60 Ala Ala Asp Glu Pro Gln Leu Leu His Gly Ala Gly Ile Cys Lys Trp 65 70 75 80 Phe Asn Val Arg Met Gly Phe Gly Phe Leu Ser Met Thr Ala Arg Ala 85 90 95 Gly Val Ala Leu Asp Pro Pro Val Asp Val Phe Val His Gln Ser Lys 100 105 110 Leu His Met Glu Gly Phe Arg Ser Leu Lys Glu Gly Glu Ala Val Glu 115 120 125 Phe Thr Phe Lys Lys Ser Ala Lys Gly Leu Glu Ser Ile Arg Val Thr 130 135 140 Gly Pro Gly Gly Val Phe Cys Ile Gly Ser Glu Arg Arg Pro Lys Gly 145 150 155 160 Lys Ser Met Gln Lys Arg Arg Ser Lys Gly Asp Arg Cys Tyr Asn Cys 165 170 175 Gly Gly Leu Asp His His Ala Lys Glu Cys Lys Leu Pro Pro Gln Pro 180 185 190 Lys Lys Cys His Phe Cys Gln Ser Ile Ser His Met Val Ala Ser Cys 195 200 205 Pro Leu Lys Ala Gln Gln Gly Pro Ser Ala Gln Gly Lys Pro Thr Tyr 210 215 220 Phe Arg Glu Glu Glu Glu Glu Ile His Ser Pro Thr Leu Leu Pro Glu 225 230 235 240 Ala Gln Asn Leu Val Ala Ala Leu Leu Ala Val Leu 245 250 <210> 181 <211> 729 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Lin28 cDNA Sequence <400> 181 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgggctccgt gtccaaccag 120 cagtttgcag gtggctgcgc caaggcggca gaagaggcgc ccgaggaggc gccggaggac 180 gcggcccggg cggcggacga gcctcagctg ctgcacggtg cgggcatctg taagtggttc 240 aacgtgcgca tggggttcgg cttcctgtcc atgaccgccc gcgccggggt cgcgctcgac 300 cccccagtgg atgtctttgt gcaccagagt aagctgcaca tggaagggtt ccggagcttg 360 aaggagggtg aggcagtgga gttcaccttt aagaagtcag ccaagggtct ggaatccatc 420 cgtgtcaccg gacctggtgg agtattctgt attgggagtg agaggcggcc aaaaggaaag 480 agcatgcaga agcgcagatc aaaaggagac aggtgctaca actgtggagg tctagatcat 540 catgccaagg aatgcaagct gccaccccag cccaagaagt gccacttctg ccagagcatc 600 agccatatgg tagcctcatg tccgctgaag gcccagcagg gccctagtgc acagggaaag 660 ccaacctact ttcgagagga agaagaagaa atccacagcc ctaccctgct cccggaggca 720 cagaattga 729 <210> 182 <211> 242 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Lin28 Amino Acid Sequence <400> 182 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Gly Ser Val Ser Asn Gln Gln Phe Ala Gly Gly Cys Ala Lys 35 40 45 Ala Ala Glu Glu Ala Pro Glu Glu Ala Pro Glu Asp Ala Ala Arg Ala 50 55 60 Ala Asp Glu Pro Gln Leu Leu His Gly Ala Gly Ile Cys Lys Trp Phe 65 70 75 80 Asn Val Arg Met Gly Phe Gly Phe Leu Ser Met Thr Ala Arg Ala Gly 85 90 95 Val Ala Leu Asp Pro Pro Val Asp Val Phe Val His Gln Ser Lys Leu 100 105 110 His Met Glu Gly Phe Arg Ser Leu Lys Glu Gly Glu Ala Val Glu Phe 115 120 125 Thr Phe Lys Lys Ser Ala Lys Gly Leu Glu Ser Ile Arg Val Thr Gly 130 135 140 Pro Gly Gly Val Phe Cys Ile Gly Ser Glu Arg Arg Pro Lys Gly Lys 145 150 155 160 Ser Met Gln Lys Arg Arg Ser Lys Gly Asp Arg Cys Tyr Asn Cys Gly 165 170 175 Gly Leu Asp His His Ala Lys Glu Cys Lys Leu Pro Pro Gln Pro Lys 180 185 190 Lys Cys His Phe Cys Gln Ser Ile Ser His Met Val Ala Ser Cys Pro 195 200 205 Leu Lys Ala Gln Gln Gly Pro Ser Ala Gln Gly Lys Pro Thr Tyr Phe 210 215 220 Arg Glu Glu Glu Glu Glu Ile His Ser Pro Thr Leu Leu Pro Glu Ala 225 230 235 240 Gln Asn <210> 183 <211> 729 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-Lin28-JO-86 MTD cDNA Sequence <400> 183 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaaggg ctccgtgtcc aaccagcagt ttgcaggtgg ctgcgccaag 120 gcggcagaag aggcgcccga ggaggcgccg gaggacgcgg cccgggcggc ggacgagcct 180 cagctgctgc acggtgcggg catctgtaag tggttcaacg tgcgcatggg gttcggcttc 240 ctgtccatga ccgcccgcgc cggggtcgcg ctcgaccccc cagtggatgt ctttgtgcac 300 cagagtaagc tgcacatgga agggttccgg agcttgaagg agggtgaggc agtggagttc 360 acctttaaga agtcagccaa gggtctggaa tccatccgtg tcaccggacc tggtggagta 420 ttctgtattg ggagtgagag gcggccaaaa ggaaagagca tgcagaagcg cagatcaaaa 480 ggagacaggt gctacaactg tggaggtcta gatcatcatg ccaaggaatg caagctgcca 540 ccccagccca agaagtgcca cttctgccag agcatcagcc atatggtagc ctcatgtccg 600 ctgaaggccc agcagggccc tagtgcacag ggaaagccaa cctactttcg agaggaagaa 660 gaagaaatcc acagccctac cctgctcccg gaggcacaga atctggcggt gctggcggcg 720 gcgccgtga 729 <210> 184 <211> 242 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-Lin28-JO-86 MTD Amino Acid Sequence <400> 184 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Gly Ser Val Ser Asn Gln 20 25 30 Gln Phe Ala Gly Gly Cys Ala Lys Ala Ala Glu Glu Ala Pro Glu Glu 35 40 45 Ala Pro Glu Asp Ala Ala Arg Ala Ala Asp Glu Pro Gln Leu Leu His 50 55 60 Gly Ala Gly Ile Cys Lys Trp Phe Asn Val Arg Met Gly Phe Gly Phe 65 70 75 80 Leu Ser Met Thr Ala Arg Ala Gly Val Ala Leu Asp Pro Pro Val Asp 85 90 95 Val Phe Val His Gln Ser Lys Leu His Met Glu Gly Phe Arg Ser Leu 100 105 110 Lys Glu Gly Glu Ala Val Glu Phe Thr Phe Lys Lys Ser Ala Lys Gly 115 120 125 Leu Glu Ser Ile Arg Val Thr Gly Pro Gly Gly Val Phe Cys Ile Gly 130 135 140 Ser Glu Arg Arg Pro Lys Gly Lys Ser Met Gln Lys Arg Arg Ser Lys 145 150 155 160 Gly Asp Arg Cys Tyr Asn Cys Gly Gly Leu Asp His His Ala Lys Glu 165 170 175 Cys Lys Leu Pro Pro Gln Pro Lys Lys Cys His Phe Cys Gln Ser Ile 180 185 190 Ser His Met Val Ala Ser Cys Pro Leu Lys Ala Gln Gln Gly Pro Ser 195 200 205 Ala Gln Gly Lys Pro Thr Tyr Phe Arg Glu Glu Glu Glu Glu Ile His 210 215 220 Ser Pro Thr Leu Leu Pro Glu Ala Gln Asn Leu Ala Val Leu Ala Ala 225 230 235 240 Ala Pro <210> 185 <211> 753 <212> DNA <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Lin28-JO-86 MTD cDNA Sequence <400> 185 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat 60 atgaagaaga agaggaagct ggcggtgctg gcggcggcgc cgggctccgt gtccaaccag 120 cagtttgcag gtggctgcgc caaggcggca gaagaggcgc ccgaggaggc gccggaggac 180 gcggcccggg cggcggacga gcctcagctg ctgcacggtg cgggcatctg taagtggttc 240 aacgtgcgca tggggttcgg cttcctgtcc atgaccgccc gcgccggggt cgcgctcgac 300 cccccagtgg atgtctttgt gcaccagagt aagctgcaca tggaagggtt ccggagcttg 360 aaggagggtg aggcagtgga gttcaccttt aagaagtcag ccaagggtct ggaatccatc 420 cgtgtcaccg gacctggtgg agtattctgt attgggagtg agaggcggcc aaaaggaaag 480 agcatgcaga agcgcagatc aaaaggagac aggtgctaca actgtggagg tctagatcat 540 catgccaagg aatgcaagct gccaccccag cccaagaagt gccacttctg ccagagcatc 600 agccatatgg tagcctcatg tccgctgaag gcccagcagg gccctagtgc acagggaaag 660 ccaacctact ttcgagagga agaagaagaa atccacagcc ctaccctgct cccggaggca 720 cagaatctgg cggtgctggc ggcggcgccg tga 753 <210> 186 <211> 250 <212> PRT <213> Artificial Sequence <220> <223> His-NLS-JO-86 MTD-Lin28-JO-86 MTD Amino Acid Sequence <400> 186 Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 1 5 10 15 Arg Gly Ser His Met Lys Lys Lys Arg Lys Leu Ala Val Leu Ala Ala 20 25 30 Ala Pro Gly Ser Val Ser Asn Gln Gln Phe Ala Gly Gly Cys Ala Lys 35 40 45 Ala Ala Glu Glu Ala Pro Glu Glu Ala Pro Glu Asp Ala Ala Arg Ala 50 55 60 Ala Asp Glu Pro Gln Leu Leu His Gly Ala Gly Ile Cys Lys Trp Phe 65 70 75 80 Asn Val Arg Met Gly Phe Gly Phe Leu Ser Met Thr Ala Arg Ala Gly 85 90 95 Val Ala Leu Asp Pro Pro Val Asp Val Phe Val His Gln Ser Lys Leu 100 105 110 His Met Glu Gly Phe Arg Ser Leu Lys Glu Gly Glu Ala Val Glu Phe 115 120 125 Thr Phe Lys Lys Ser Ala Lys Gly Leu Glu Ser Ile Arg Val Thr Gly 130 135 140 Pro Gly Gly Val Phe Cys Ile Gly Ser Glu Arg Arg Pro Lys Gly Lys 145 150 155 160 Ser Met Gln Lys Arg Arg Ser Lys Gly Asp Arg Cys Tyr Asn Cys Gly 165 170 175 Gly Leu Asp His His Ala Lys Glu Cys Lys Leu Pro Pro Gln Pro Lys 180 185 190 Lys Cys His Phe Cys Gln Ser Ile Ser His Met Val Ala Ser Cys Pro 195 200 205 Leu Lys Ala Gln Gln Gly Pro Ser Ala Gln Gly Lys Pro Thr Tyr Phe 210 215 220 Arg Glu Glu Glu Glu Glu Ile His Ser Pro Thr Leu Leu Pro Glu Ala 225 230 235 240 Gln Asn Leu Ala Val Leu Ala Ala Ala Pro 245 250 <210> 187 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> HhN-5' <400> 187 ccgcatatga agaagaagag gaagagtgtg gatccagctt gtccc 45 <210> 188 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> HhN-3' <400> 188 ccgcatatgt cacacgtctt caggttgcat gttcat 36 <210> 189 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> HM1hN-5' <400> 189 ccgcatatga agaagaagag gaagctggtg gcggcgctgc tggcggtgct gagtgtggat 60 ccagcttgtc cccaa 75 <210> 190 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> HhNM1-3' <400> 190 ccgcatatgt cacagcaccg ccagcagcgc cgccaccagc acgtcttcag gttgcatgtt 60 cat 63 <210> 191 <211> 72 <212> DNA <213> Artificial Sequence <220> <223> HM2hN-5' <400> 191 ccgcatatga agaagaagag gaagctggcg gtgctggcgg cggcgccgag tgtggatcca 60 gcttgtcccc aa 72 <210> 192 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> HhNM2-3' <400> 192 ccgcatatgt cacggcgccg ccgccagcac cgccagcacg tcttcaggtt gcatgttcat 60 60 <210> 193 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> HhO-5' <400> 193 ccgcatatga agaagaagag gaaggcggga cacctggctt cggat 45 <210> 194 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> HhO-3' <400> 194 ccgcatatgt cagtttgaat gcatgggaga gcccag 36 <210> 195 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> HM1hO-5' <400> 195 ccgcatatga agaagaagag gaagctggtg gcggcgctgc tggcggtgct ggcgggacac 60 ctggcttcgg atttc 75 <210> 196 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> HhOM1-3' <400> 196 ccgcatatgt cacagcaccg ccagcagcgc cgccaccagg tttgaatgca tgggagagcc 60 cag 63 <210> 197 <211> 72 <212> DNA <213> Artificial Sequence <220> <223> HM2hO-5' <400> 197 ccgcatatga agaagaagag gaagctggcg gtgctggcgg cggcgccggc gggacacctg 60 gcttcggatt tc 72 <210> 198 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> HhOM2-3' <400> 198 ccgcatatgt cacggcgccg ccgccagcac cgccaggttt gaatgcatgg gagagcccag 60 60 <210> 199 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> HhS-5' <400> 199 ccgcatatga agaagaagag gaagtacaac atgatggaga cggag 45 <210> 200 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> HhS-3' <400> 200 ccgcatatgt cacatgtgtg agaggggcag tgtgcc 36 <210> 201 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> HM1hS-5' <400> 201 ccgcatatga agaagaagag gaagctggtg gcggcgctgc tggcggtgct gtacaacatg 60 atggagacgg agctg 75 <210> 202 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> HhSM1-3' <400> 202 ccgcatatgt cacagcaccg ccagcagcgc cgccaccagc atgtgtgaga ggggcagtgt 60 gcc 63 <210> 203 <211> 72 <212> DNA <213> Artificial Sequence <220> <223> HM2hS-5' <400> 203 ccgcatatga agaagaagag gaagctggcg gtgctggcgg cggcgccgta caacatgatg 60 gagacggagc tg 72 <210> 204 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> HhSM2-3' <400> 204 ccgcatatgt cacggcgccg ccgccagcac cgccagcatg tgtgagaggg gcagtgtgcc 60 60 <210> 205 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> HhK-5' <400> 205 ccgcatatga agaagaagag gaaggctgtc agcgacgcgc tgctc 45 <210> 206 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> HhK-3' <400> 206 ccgcatatgt taaaaatgcc tcttcatgtg taaggc 36 <210> 207 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> HM1hK-5' <400> 207 ccgcatatga agaagaagag gaagctggtg gcggcgctgc tggcggtgct ggctgtcagc 60 gacgcgctgc tccca 75 <210> 208 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> HhKM1-3' <400> 208 ccgcatatgt tacagcaccg ccagcagcgc cgccaccaga aaatgcctct tcatgtgtaa 60 ggc 63 <210> 209 <211> 72 <212> DNA <213> Artificial Sequence <220> <223> HM2hK-5' <400> 209 ccgcatatga agaagaagag gaagctggcg gtgctggcgg cggcgccggc tgtcagcgac 60 gcgctgctcc ca 72 <210> 210 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> HhKM2-3' <400> 210 ccgcatatgt tacggcgccg ccgccagcac cgccagaaaa tgcctcttca tgtgtaaggc 60 60 <210> 211 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> HhM-5' <400> 211 ccgcatatga agaagaagag gaaggatttt tttcgggtag tggaa 45 <210> 212 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> HhM-3' <400> 212 ccgcatatgt tacgcacaag agttccgtag ctgttc 36 <210> 213 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> HM1hM-5' <400> 213 ccgcatatga agaagaagag gaagctggtg gcggcgctgc tggcggtgct ggattttttt 60 cgggtagtgg aaaac 75 <210> 214 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> HhMM1-3' <400> 214 ccgcatatgt tacagcaccg ccagcagcgc cgccaccagc gcacaagagt tccgtagctg 60 ttc 63 <210> 215 <211> 72 <212> DNA <213> Artificial Sequence <220> <223> HM2hM-5' <400> 215 ccgcatatga agaagaagag gaagctggcg gtgctggcgg cggcgccgga tttttttcgg 60 gtagtggaaa ac 72 <210> 216 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> HhMM2-3' <400> 216 ccgcatatgt tacggcgccg ccgccagcac cgccagcgca caagagttcc gtagctgttc 60 60 <210> 217 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> HhL-5' <400> 217 ccgggatcca agaagaagag gaagggctcc gtgtccaacc agcag 45 <210> 218 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> HhL-3' <400> 218 ccgggatcct caattctgag cctccgggag cagggt 36 <210> 219 <211> 75 <212> DNA <213> Artificial Sequence <220> <223> HM1hL-5' <400> 219 ccgggatcca agaagaagag gaagctggtg gcggcgctgc tggcggtgct gggctccgtg 60 tccaaccagc agttt 75 <210> 220 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> HhLM1-3' <400> 220 ccgggatcct tacagcaccg ccagcagcgc cgccaccagt caattctgag cctccgggag 60 cag 63 <210> 221 <211> 72 <212> DNA <213> Artificial Sequence <220> <223> HM2hL-5' <400> 221 ccgggatcca agaagaagag gaagctggcg gtgctggcgg cggcgccggg ctccgtgtcc 60 aaccagcagt tt 72 <210> 222 <211> 60 <212> DNA <213> Artificial Sequence <220> <223> HhLM2-3' <400> 222 ccgggatcct tacggcgccg ccgccagcac cgccagtcaa ttctgagcct ccgggagcag 60 60 <210> 223 <211> 48 <212> DNA <213> Artificial Sequence <220> <223> JO-10 MTD cDNA Sequence <400> 223 ctgggcggcg cggtggtggc ggcgccggtg gcggcggcgg tggcgccg 48 <210> 224 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> JO-10 MTD Amino Acid Sequence Sequence <400> 224 Leu Gly Gly Ala Val Val Ala Ala Pro Val Ala Ala Ala Val Ala Pro 1 5 10 15 <210> 225 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> JO-52 MTD cDNA Sequence <400> 225 ccgctgctgc tgctgctgcc ggcgctg 27 <210> 226 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> JO-52 MTD Amino Acid Sequence Sequence <400> 226 Pro Leu Leu Leu Leu Leu Pro Ala Leu 1 5 <210> 227 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> JO-132 MTD cDNA Sequence <400> 227 gcggtggtgg tgccggcgat tgtgctggcg gcgccg 36 <210> 228 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> JO-132 MTD Amino Acid Sequence Sequence <400> 228 Ala Val Val Val Pro Ala Ile Val Leu Ala Ala Pro 1 5 10 <210> 229 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> JO-145 MTD cDNA Sequence <400> 229 gcggcggcgc cggtgctgct gctgctgctg 30 <210> 230 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> JO-145 MTD Amino Acid Sequence Sequence <400> 230 Ala Ala Ala Pro Val Leu Leu Leu Leu Leu 1 5 10 <210> 231 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> JO-173 MTD cDNA Sequence <400> 231 gcggtgattc cgattctggc ggtgccg 27 <210> 232 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> JO-173 MTD Amino Acid Sequence Sequence <400> 232 Ala Val Ile Pro Ile Leu Ala Val Pro 1 5 <210> 233 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> JO-181 MTD cDNA Sequence <400> 233 gcggtgctgc tgctgccggc ggcggcg 27 <210> 234 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> JO-181 MTD Amino Acid Sequence Sequence <400> 234 Ala Val Leu Leu Leu Pro Ala Ala Ala 1 5 <210> 235 <211> 960 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-Nanog cDNA Sequence <400> 235 atgggttcaa gtcatcatca tcatcatcat aaaaaaaaac gcaaaagtgt ggatccggca 60 tgcccgcagt cactgccgtg ttttgaagcg tcggactgca aagaaagctc tccgatgccg 120 gttatttgtg gcccggaaga aaactatccg tcactgcaga tgagttccgc cgaaatgccg 180 cataccgaaa cggtcagccc gctgccgtca tcgatggatc tgctgatcca agattcaccg 240 gacagctcta cgtcgccgaa aggtaaacag ccgaccagtg cagaaaactc cgttgctaaa 300 aaagaagata aagtgccggt taaaaaacaa aaaacccgta cggtctttag ttccacgcag 360 ctgtgcgtgc tgaatgaccg tttccagcgc caaaaatatc tgagcctgca gcaaatgcaa 420 gaactgagca acattctgaa tctgtcttac aaacaggtga aaacctggtt ccagaaccaa 480 cgtatgaaaa gtaaacgctg gcagaaaaac aattggccga aaaactccaa tggcgttacg 540 cagaaagcga gtgccccgac ctacccgtcc ctgtattcat cgtaccatca gggctgtctg 600 gtcaacccga ccggtaatct gccgatgtgg tctaatcaga cctggaacaa tagtacgtgg 660 tccaaccaga cccaaaatat tcagagctgg tctaaccaca gctggaatac ccagacgtgg 720 tgcacccaaa gctggaacaa tcaggcatgg aactctccgt tttataattg tggtgaagaa 780 tcactgcagt cgtgcatgca gttccaaccg aacagcccgg cttctgatct ggaagcggcc 840 ctggaagcag ctggcgaagg tctgaatgtt atccagcaaa ccacgcgcta ctttagcacc 900 ccgcaaacga tggacctgtt tctgaattac tcgatgaata tgcaaccgga agatgtgtaa 960 960 <210> 236 <211> 1008 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-10 MTD Nanog cDNA Sequence <400> 236 atgggttcaa gtcatcatca tcatcatcat aaaaaaaaac gcaaactggg cggcgcggtc 60 gtggcggctc cggtggcagc agcggtggca ccgagtgtgg atccggcatg cccgcagtca 120 ctgccgtgtt ttgaagcgtc ggactgcaaa gaaagctctc cgatgccggt tatttgtggc 180 ccggaagaaa actatccgtc actgcagatg agttccgccg aaatgccgca taccgaaacg 240 gtcagcccgc tgccgtcatc gatggatctg ctgatccaag attcaccgga cagctctacg 300 tcgccgaaag gtaaacagcc gaccagtgca gaaaactccg ttgctaaaaa agaagataaa 360 gtgccggtta aaaaacaaaa aacccgtacg gtctttagtt ccacgcagct gtgcgtgctg 420 aatgaccgtt tccagcgcca aaaatatctg agcctgcagc aaatgcaaga actgagcaac 480 attctgaatc tgtcttacaa acaggtgaaa acctggttcc agaaccaacg tatgaaaagt 540 aaacgctggc agaaaaacaa ttggccgaaa aactccaatg gcgttacgca gaaagcgagt 600 gccccgacct acccgtccct gtattcatcg taccatcagg gctgtctggt caacccgacc 660 ggtaatctgc cgatgtggtc taatcagacc tggaacaata gtacgtggtc caaccagacc 720 caaaatattc agagctggtc taaccacagc tggaataccc agacgtggtg cacccaaagc 780 tggaacaatc aggcatggaa ctctccgttt tataattgtg gtgaagaatc actgcagtcg 840 tgcatgcagt tccaaccgaa cagcccggct tctgatctgg aagcggccct ggaagcagct 900 ggcgaaggtc tgaatgttat ccagcaaacc acgcgctact ttagcacccc gcaaacgatg 960 gacctgtttc tgaattactc gatgaatatg caaccggaag atgtgtaa 1008 <210> 237 <211> 335 <212> PRT <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-10 MTD Nanog Amino Acid Sequence <400> 237 Met Gly Ser Ser His His His His His His Lys Lys Lys Arg Lys Leu 1 5 10 15 Gly Gly Ala Val Val Ala Ala Pro Val Ala Ala Ala Val Ala Pro Ser 20 25 30 Val Asp Pro Ala Cys Pro Gln Ser Leu Pro Cys Phe Glu Ala Ser Asp 35 40 45 Cys Lys Glu Ser Ser Pro Met Pro Val Ile Cys Gly Pro Glu Glu Asn 50 55 60 Tyr Pro Ser Leu Gln Met Ser Ser Ala Glu Met Pro His Thr Glu Thr 65 70 75 80 Val Ser Pro Leu Pro Ser Ser Met Asp Leu Leu Ile Gln Asp Ser Pro 85 90 95 Asp Ser Ser Thr Ser Pro Lys Gly Lys Gln Pro Thr Ser Ala Glu Asn 100 105 110 Ser Val Ala Lys Lys Glu Asp Lys Val Pro Val Lys Lys Gln Lys Thr 115 120 125 Arg Thr Val Phe Ser Ser Thr Gln Leu Cys Val Leu Asn Asp Arg Phe 130 135 140 Gln Arg Gln Lys Tyr Leu Ser Leu Gln Gln Met Gln Glu Leu Ser Asn 145 150 155 160 Ile Leu Asn Leu Ser Tyr Lys Gln Val Lys Thr Trp Phe Gln Asn Gln 165 170 175 Arg Met Lys Ser Lys Arg Trp Gln Lys Asn Asn Trp Pro Lys Asn Ser 180 185 190 Asn Gly Val Thr Gln Lys Ala Ser Ala Pro Thr Tyr Pro Ser Leu Tyr 195 200 205 Ser Ser Tyr His Gln Gly Cys Leu Val Asn Pro Thr Gly Asn Leu Pro 210 215 220 Met Trp Ser Asn Gln Thr Trp Asn Asn Ser Thr Trp Ser Asn Gln Thr 225 230 235 240 Gln Asn Ile Gln Ser Trp Ser Asn His Ser Trp Asn Thr Gln Thr Trp 245 250 255 Cys Thr Gln Ser Trp Asn Asn Gln Ala Trp Asn Ser Pro Phe Tyr Asn 260 265 270 Cys Gly Glu Glu Ser Leu Gln Ser Cys Met Gln Phe Gln Pro Asn Ser 275 280 285 Pro Ala Ser Asp Leu Glu Ala Ala Leu Glu Ala Ala Gly Glu Gly Leu 290 295 300 Asn Val Ile Gln Gln Thr Thr Arg Tyr Phe Ser Thr Pro Gln Thr Met 305 310 315 320 Asp Leu Phe Leu Asn Tyr Ser Met Asn Met Gln Pro Glu Asp Val 325 330 335 <210> 238 <211> 1125 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-Oct4 cDNA Sequence <400> 238 atgggttcgt cgcatcatca tcatcatcat aaaaaaaaac gcaaagctgg tcacctggct 60 tccgattttg ctttctcacc gccgccgggc ggtggcggtg atggtccggg cggtccggaa 120 ccgggttggg tggacccgcg tacctggctg tccttccagg gtccgccggg cggtccgggt 180 attggtccgg gtgttggtcc gggctcagaa gtctggggta tcccgccgtg cccgccgccg 240 tatgaatttt gcggtggtat ggcatactgt ggtccgcagg tcggtgtggg tctggttccg 300 cagggcggtc tggaaacgtc gcaaccggaa ggcgaagcag gtgttggcgt cgaatcgaac 360 agcgatggcg ctagcccgga accgtgtacc gtgacgccgg gtgcggttaa actggaaaaa 420 gaaaaactgg aacagaatcc ggaagaaagc caagatatta aagcgctgca gaaagaactg 480 gaacaattcg ccaaactgct gaaacagaaa cgcatcaccc tgggttatac gcaagcggac 540 gttggtctga ccctgggcgt cctgtttggt aaagtgttct cgcagaccac gatttgccgc 600 tttgaagctc tgcaactgag cttcaaaaac atgtgtaaac tgcgtccgct gctgcagaaa 660 tgggtcgaag aagcggataa caatgaaaat ctgcaggaaa tttgcaaagc agaaaccctg 720 gtgcaagctc gtaaacgcaa acgtacgtct atcgaaaacc gcgttcgtgg caacctggaa 780 aatctgttcc tgcagtgccc gaaaccgacc ctgcagcaaa ttagccatat cgcccagcaa 840 ctgggcctgg aaaaagacgt ggttcgtgtg tggttttgta atcgtcgcca gaaaggtaaa 900 cgcagctcta gtgattacgc acagcgtgaa gactttgaag cagcaggttc tccgttcagt 960 ggcggtccgg tctcttttcc gctggcaccg ggtccgcatt ttggtacccc gggttatggc 1020 agtccgcact tcacggcact gtactcctca gtgccgtttc cggaaggcga agcgttcccg 1080 ccggtctccg tcaccaccct gggtagtccg atgcacagta attaa 1125 <210> 239 <211> 1152 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-52 MTD Oct4 cDNA Sequence <400> 239 atgggttcgt cgcatcatca tcatcatcat aaaaaaaaac gcaaaccgct gctgctgctg 60 ctgccggccc tggctggtca cctggcttcc gattttgctt tctcaccgcc gccgggcggt 120 ggcggtgatg gtccgggcgg tccggaaccg ggttgggtgg acccgcgtac ctggctgtcc 180 ttccagggtc cgccgggcgg tccgggtatt ggtccgggtg ttggtccggg ctcagaagtc 240 tggggtatcc cgccgtgccc gccgccgtat gaattttgcg gtggtatggc atactgtggt 300 ccgcaggtcg gtgtgggtct ggttccgcag ggcggtctgg aaacgtcgca accggaaggc 360 gaagcaggtg ttggcgtcga atcgaacagc gatggcgcta gcccggaacc gtgtaccgtg 420 acgccgggtg cggttaaact ggaaaaagaa aaactggaac agaatccgga agaaagccaa 480 gatattaaag cgctgcagaa agaactggaa caattcgcca aactgctgaa acagaaacgc 540 atcaccctgg gttatacgca agcggacgtt ggtctgaccc tgggcgtcct gtttggtaaa 600 gtgttctcgc agaccacgat ttgccgcttt gaagctctgc aactgagctt caaaaacatg 660 tgtaaactgc gtccgctgct gcagaaatgg gtcgaagaag cggataacaa tgaaaatctg 720 caggaaattt gcaaagcaga aaccctggtg caagctcgta aacgcaaacg tacgtctatc 780 gaaaaccgcg ttcgtggcaa cctggaaaat ctgttcctgc agtgcccgaa accgaccctg 840 cagcaaatta gccatatcgc ccagcaactg ggcctggaaa aagacgtggt tcgtgtgtgg 900 ttttgtaatc gtcgccagaa aggtaaacgc agctctagtg attacgcaca gcgtgaagac 960 tttgaagcag caggttctcc gttcagtggc ggtccggtct cttttccgct ggcaccgggt 1020 ccgcattttg gtaccccggg ttatggcagt ccgcacttca cggcactgta ctcctcagtg 1080 ccgtttccgg aaggcgaagc gttcccgccg gtctccgtca ccaccctggg tagtccgatg 1140 cacagtaatt aa 1152 <210> 240 <211> 383 <212> PRT <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-52 MTD Oct4 Amino Acid Sequence <400> 240 Met Gly Ser Ser His His His His His His Lys Lys Lys Arg Lys Pro 1 5 10 15 Leu Leu Leu Leu Leu Pro Ala Leu Ala Gly His Leu Ala Ser Asp Phe 20 25 30 Ala Phe Ser Pro Pro Pro Gly Gly Gly Gly Asp Gly Pro Gly Gly Pro 35 40 45 Glu Pro Gly Trp Val Asp Pro Arg Thr Trp Leu Ser Phe Gln Gly Pro 50 55 60 Pro Gly Gly Pro Gly Ile Gly Pro Gly Val Gly Pro Gly Ser Glu Val 65 70 75 80 Trp Gly Ile Pro Pro Cys Pro Pro Pro Tyr Glu Phe Cys Gly Gly Met 85 90 95 Ala Tyr Cys Gly Pro Gln Val Gly Val Gly Leu Val Pro Gln Gly Gly 100 105 110 Leu Glu Thr Ser Gln Pro Glu Gly Glu Ala Gly Val Gly Val Glu Ser 115 120 125 Asn Ser Asp Gly Ala Ser Pro Glu Pro Cys Thr Val Thr Pro Gly Ala 130 135 140 Val Lys Leu Glu Lys Glu Lys Leu Glu Gln Asn Pro Glu Glu Ser Gln 145 150 155 160 Asp Ile Lys Ala Leu Gln Lys Glu Leu Glu Gln Phe Ala Lys Leu Leu 165 170 175 Lys Gln Lys Arg Ile Thr Leu Gly Tyr Thr Gln Ala Asp Val Gly Leu 180 185 190 Thr Leu Gly Val Leu Phe Gly Lys Val Phe Ser Gln Thr Thr Ile Cys 195 200 205 Arg Phe Glu Ala Leu Gln Leu Ser Phe Lys Asn Met Cys Lys Leu Arg 210 215 220 Pro Leu Leu Gln Lys Trp Val Glu Glu Ala Asp Asn Asn Glu Asn Leu 225 230 235 240 Gln Glu Ile Cys Lys Ala Glu Thr Leu Val Gln Ala Arg Lys Arg Lys 245 250 255 Arg Thr Ser Ile Glu Asn Arg Val Arg Gly Asn Leu Glu Asn Leu Phe 260 265 270 Leu Gln Cys Pro Lys Pro Thr Leu Gln Gln Ile Ser His Ile Ala Gln 275 280 285 Gln Leu Gly Leu Glu Lys Asp Val Val Arg Val Trp Phe Cys Asn Arg 290 295 300 Arg Gln Lys Gly Lys Arg Ser Ser Ser Asp Tyr Ala Gln Arg Glu Asp 305 310 315 320 Phe Glu Ala Ala Gly Ser Pro Phe Ser Gly Gly Pro Val Ser Phe Pro 325 330 335 Leu Ala Pro Gly Pro His Phe Gly Thr Pro Gly Tyr Gly Ser Pro His 340 345 350 Phe Thr Ala Leu Tyr Ser Ser Val Pro Phe Pro Glu Gly Glu Ala Phe 355 360 365 Pro Pro Val Ser Val Thr Thr Leu Gly Ser Pro Met His Ser Asn 370 375 380 <210> 241 <211> 996 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-Sox2 cDNA Sequence <400> 241 atgggctcgt cgcaccacca ccaccaccac aaaaaaaaac gcaaatataa catgatggaa 60 accgaactga aaccgccggg tccgcagcaa acctcgggcg gtggcggtgg caacagcacg 120 gcggcggcgg cgggtggcaa ccagaaaaat tcaccggatc gtgtgaaacg cccgatgaat 180 gcatttatgg tttggagtcg tggtcagcgt cgcaaaatgg ctcaagaaaa cccgaaaatg 240 cataactctg aaatcagtaa acgtctgggc gcggaatgga aactgctgag cgaaaccgaa 300 aaacgcccgt tcatcgatga agcgaaacgt ctgcgcgccc tgcatatgaa agaacacccg 360 gactataaat accgcccgcg tcgcaaaacg aaaacgctga tgaaaaaaga taaatatacg 420 ctgccgggtg gcctgctggc accgggtggc aacagcatgg cgtcaggtgt tggtgtcggt 480 gcaggtctgg gtgcaggtgt caatcagcgt atggattctt acgcccatat gaacggttgg 540 agtaatggca gttattccat gatgcaggac caactgggtt acccgcagca tccgggtctg 600 aacgcacacg gcgcagcaca gatgcaaccg atgcaccgct atgacgtttc cgccctgcag 660 tacaactcaa tgaccagctc tcaaacgtat atgaatggct ctccgaccta ttcaatgtcg 720 tacagccagc agggtacgcc gggtatggca ctgggttcga tgggcagcgt ggttaaaagc 780 gaagccagtt cctcaccgcc ggtcgtgacc tcgagctctc actcccgtgc accgtgccag 840 gctggtgatc tgcgcgacat gattagcatg tatctgccgg gtgcagaagt gccggaaccg 900 gcagctccgt ctcgtctgca tatgagtcag cactatcaat cgggtccggt tccgggtacg 960 gctatcaacg gtacgctgcc gctgtcgcac atgtaa 996 <210> 242 <211> 1023 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-181 MTD Sox2 cDNA Sequence <400> 242 atgggctcgt cgcaccacca ccaccaccac aaaaaaaaac gcaaagcggt cctgctgctg 60 ccggctgctg cctataacat gatggaaacc gaactgaaac cgccgggtcc gcagcaaacc 120 tcgggcggtg gcggtggcaa cagcacggcg gcggcggcgg gtggcaacca gaaaaattca 180 ccggatcgtg tgaaacgccc gatgaatgca tttatggttt ggagtcgtgg tcagcgtcgc 240 aaaatggctc aagaaaaccc gaaaatgcat aactctgaaa tcagtaaacg tctgggcgcg 300 gaatggaaac tgctgagcga aaccgaaaaa cgcccgttca tcgatgaagc gaaacgtctg 360 cgcgccctgc atatgaaaga acacccggac tataaatacc gcccgcgtcg caaaacgaaa 420 acgctgatga aaaaagataa atatacgctg ccgggtggcc tgctggcacc gggtggcaac 480 agcatggcgt caggtgttgg tgtcggtgca ggtctgggtg caggtgtcaa tcagcgtatg 540 gattcttacg cccatatgaa cggttggagt aatggcagtt attccatgat gcaggaccaa 600 ctgggttacc cgcagcatcc gggtctgaac gcacacggcg cagcacagat gcaaccgatg 660 caccgctatg acgtttccgc cctgcagtac aactcaatga ccagctctca aacgtatatg 720 aatggctctc cgacctattc aatgtcgtac agccagcagg gtacgccggg tatggcactg 780 ggttcgatgg gcagcgtggt taaaagcgaa gccagttcct caccgccggt cgtgacctcg 840 agctctcact cccgtgcacc gtgccaggct ggtgatctgc gcgacatgat tagcatgtat 900 ctgccgggtg cagaagtgcc ggaaccggca gctccgtctc gtctgcatat gagtcagcac 960 tatcaatcgg gtccggttcc gggtacggct atcaacggta cgctgccgct gtcgcacatg 1020 taa 1023 <210> 243 <211> 340 <212> PRT <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-181 MTD Sox2 Amino Acid Sequence <400> 243 Met Gly Ser Ser His His His His His His Lys Lys Lys Arg Lys Ala 1 5 10 15 Val Leu Leu Leu Pro Ala Ala Ala Tyr Asn Met Met Glu Thr Glu Leu 20 25 30 Lys Pro Pro Gly Pro Gln Gln Thr Ser Gly Gly Gly Gly Gly Asn Ser 35 40 45 Thr Ala Ala Ala Ala Gly Gly Asn Gln Lys Asn Ser Pro Asp Arg Val 50 55 60 Lys Arg Pro Met Asn Ala Phe Met Val Trp Ser Arg Gly Gln Arg Arg 65 70 75 80 Lys Met Ala Gln Glu Asn Pro Lys Met His Asn Ser Glu Ile Ser Lys 85 90 95 Arg Leu Gly Ala Glu Trp Lys Leu Leu Ser Glu Thr Glu Lys Arg Pro 100 105 110 Phe Ile Asp Glu Ala Lys Arg Leu Arg Ala Leu His Met Lys Glu His 115 120 125 Pro Asp Tyr Lys Tyr Arg Pro Arg Arg Lys Thr Lys Thr Leu Met Lys 130 135 140 Lys Asp Lys Tyr Thr Leu Pro Gly Gly Leu Leu Ala Pro Gly Gly Asn 145 150 155 160 Ser Met Ala Ser Gly Val Gly Val Gly Ala Gly Leu Gly Ala Gly Val 165 170 175 Asn Gln Arg Met Asp Ser Tyr Ala His Met Asn Gly Trp Ser Asn Gly 180 185 190 Ser Tyr Ser Met Met Gln Asp Gln Leu Gly Tyr Pro Gln His Pro Gly 195 200 205 Leu Asn Ala His Gly Ala Ala Gln Met Gln Pro Met His Arg Tyr Asp 210 215 220 Val Ser Ala Leu Gln Tyr Asn Ser Met Thr Ser Ser Gln Thr Tyr Met 225 230 235 240 Asn Gly Ser Pro Thr Tyr Ser Met Ser Tyr Ser Gln Gln Gly Thr Pro 245 250 255 Gly Met Ala Leu Gly Ser Met Gly Ser Val Val Lys Ser Glu Ala Ser 260 265 270 Ser Ser Pro Pro Val Val Thr Ser Ser Ser His Ser Arg Ala Pro Cys 275 280 285 Gln Ala Gly Asp Leu Arg Asp Met Ile Ser Met Tyr Leu Pro Gly Ala 290 295 300 Glu Val Pro Glu Pro Ala Ala Pro Ser Arg Leu His Met Ser Gln His 305 310 315 320 Tyr Gln Ser Gly Pro Val Pro Gly Thr Ala Ile Asn Gly Thr Leu Pro 325 330 335 Leu Ser His Met 340 <210> 244 <211> 1455 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-Klf4 cDNA Sequence <400> 244 atgggttcga gtcatcatca tcatcatcat aaaaaaaaac gtaaagcagt gagcgacgca 60 ctgctgccgt cgtttagcac gttcgctagt ggtccggcag gtcgtgaaaa aaccctgcgt 120 caggcaggtg caccgaacaa tcgttggcgc gaagaactgt cccacatgaa acgtctgccg 180 ccggtgctgc cgggtcgtcc gtatgatctg gcggcggcaa cggttgcaac cgacctggaa 240 tcaggcggtg ctggtgctgc atgcggtggt tcgaacctgg caccgctgcc gcgtcgcgaa 300 accgaagaat ttaacgatct gctggatctg gacttcattc tgtctaatag tctgacgcat 360 ccgccggaaa gcgtcgcagc aaccgtgagc tctagtgctt ctgcgtcctc atcgagctct 420 ccgagttcct caggtccggc atcggcaccg agcacgtgtt cttttaccta tccgatccgt 480 gcaggcaatg atccgggtgt tgctccgggc ggtaccggcg gtggcctgct gtacggtcgt 540 gaaagcgctc cgccgccgac cgcaccgttt aacctggccg atattaatga cgtttccccg 600 tcaggtggct tcgtcgcaga actgctgcgt ccggaactgg acccggtcta tatcccgccg 660 cagcaaccgc aaccgccggg tggcggtctg atgggtaaat ttgtgctgaa agcctcgctg 720 agcgcaccgg gcagcgaata tggttctccg agtgtgattt ccgtttcaaa aggcagtccg 780 gatggttccc acccggtggt tgtcgcaccg tacaacggtg gtccgccgcg tacgtgcccg 840 aaaatcaaac aagaagccgt gtcgagctgt acccatctgg gtgcaggtcc gccgctgtca 900 aatggtcacc gtccggctgc acatgatttc ccgctgggtc gtcagctgcc gtctcgtacc 960 acgccgaccc tgggtctgga agaagttctg tctagtcgtg actgccatcc ggcactgccg 1020 ctgccgccgg gctttcatcc gcacccgggt ccgaactatc cgtctttcct gccggatcag 1080 atgcaaccgc aggtcccgcc gctgcactac caggaactga tgccgccggg cagttgtatg 1140 ccggaagaac cgaaaccgaa acgtggtcgt cgctcctggc cgcgtaaacg taccgcaacg 1200 catacctgcg actatgccgg ctgtggtaaa acgtacacca aatcctcaca tctgaaagcg 1260 cacctgcgca cgcataccgg cgaaaaaccg tatcactgcg attgggacgg ctgtggttgg 1320 aaatttgccc gtagcgatga actgacgcgt cattaccgca aacataccgg tcaccgcccg 1380 ttccaatgcc agaaatgcga ccgcgccttc tcccgttccg accacctggc actgcacatg 1440 aaacgtcact tctaa 1455 <210> 245 <211> 1482 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-173 MTD Klf4 cDNA Sequence <400> 245 atgggttcga gtcatcatca tcatcatcat aaaaaaaaac gtaaagcagt catcccgatt 60 ctggcggtcc cggcagtgag cgacgcactg ctgccgtcgt ttagcacgtt cgctagtggt 120 ccggcaggtc gtgaaaaaac cctgcgtcag gcaggtgcac cgaacaatcg ttggcgcgaa 180 gaactgtccc acatgaaacg tctgccgccg gtgctgccgg gtcgtccgta tgatctggcg 240 gcggcaacgg ttgcaaccga cctggaatca ggcggtgctg gtgctgcatg cggtggttcg 300 aacctggcac cgctgccgcg tcgcgaaacc gaagaattta acgatctgct ggatctggac 360 ttcattctgt ctaatagtct gacgcatccg ccggaaagcg tcgcagcaac cgtgagctct 420 agtgcttctg cgtcctcatc gagctctccg agttcctcag gtccggcatc ggcaccgagc 480 acgtgttctt ttacctatcc gatccgtgca ggcaatgatc cgggtgttgc tccgggcggt 540 accggcggtg gcctgctgta cggtcgtgaa agcgctccgc cgccgaccgc accgtttaac 600 ctggccgata ttaatgacgt ttccccgtca ggtggcttcg tcgcagaact gctgcgtccg 660 gaactggacc cggtctatat cccgccgcag caaccgcaac cgccgggtgg cggtctgatg 720 ggtaaatttg tgctgaaagc ctcgctgagc gcaccgggca gcgaatatgg ttctccgagt 780 gtgatttccg tttcaaaagg cagtccggat ggttcccacc cggtggttgt cgcaccgtac 840 aacggtggtc cgccgcgtac gtgcccgaaa atcaaacaag aagccgtgtc gagctgtacc 900 catctgggtg caggtccgcc gctgtcaaat ggtcaccgtc cggctgcaca tgatttcccg 960 ctgggtcgtc agctgccgtc tcgtaccacg ccgaccctgg gtctggaaga agttctgtct 1020 agtcgtgact gccatccggc actgccgctg ccgccgggct ttcatccgca cccgggtccg 1080 aactatccgt ctttcctgcc ggatcagatg caaccgcagg tcccgccgct gcactaccag 1140 gaactgatgc cgccgggcag ttgtatgccg gaagaaccga aaccgaaacg tggtcgtcgc 1200 tcctggccgc gtaaacgtac cgcaacgcat acctgcgact atgccggctg tggtaaaacg 1260 tacaccaaat cctcacatct gaaagcgcac ctgcgcacgc ataccggcga aaaaccgtat 1320 cactgcgatt gggacggctg tggttggaaa tttgcccgta gcgatgaact gacgcgtcat 1380 taccgcaaac ataccggtca ccgcccgttc caatgccaga aatgcgaccg cgccttctcc 1440 cgttccgacc acctggcact gcacatgaaa cgtcacttct aa 1482 <210> 246 <211> 493 <212> PRT <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-173 MTD Klf4 Amino Acid Sequence <400> 246 Met Gly Ser Ser His His His His His His Lys Lys Lys Arg Lys Ala 1 5 10 15 Val Ile Pro Ile Leu Ala Val Pro Ala Val Ser Asp Ala Leu Leu Pro 20 25 30 Ser Phe Ser Thr Phe Ala Ser Gly Pro Ala Gly Arg Glu Lys Thr Leu 35 40 45 Arg Gln Ala Gly Ala Pro Asn Asn Arg Trp Arg Glu Glu Leu Ser His 50 55 60 Met Lys Arg Leu Pro Pro Val Leu Pro Gly Arg Pro Tyr Asp Leu Ala 65 70 75 80 Ala Ala Thr Val Ala Thr Asp Leu Glu Ser Gly Gly Ala Gly Ala Ala 85 90 95 Cys Gly Gly Ser Asn Leu Ala Pro Leu Pro Arg Arg Glu Thr Glu Glu 100 105 110 Phe Asn Asp Leu Leu Asp Leu Asp Phe Ile Leu Ser Asn Ser Leu Thr 115 120 125 His Pro Pro Glu Ser Val Ala Ala Thr Val Ser Ser Ser Ala Ser Ala 130 135 140 Ser Ser Ser Ser Ser Pro Ser Ser Ser Gly Pro Ala Ser Ala Pro Ser 145 150 155 160 Thr Cys Ser Phe Thr Tyr Pro Ile Arg Ala Gly Asn Asp Pro Gly Val 165 170 175 Ala Pro Gly Gly Thr Gly Gly Gly Leu Leu Tyr Gly Arg Glu Ser Ala 180 185 190 Pro Pro Pro Thr Ala Pro Phe Asn Leu Ala Asp Ile Asn Asp Val Ser 195 200 205 Pro Ser Gly Gly Phe Val Ala Glu Leu Leu Arg Pro Glu Leu Asp Pro 210 215 220 Val Tyr Ile Pro Pro Gln Gln Pro Gln Pro Pro Gly Gly Gly Leu Met 225 230 235 240 Gly Lys Phe Val Leu Lys Ala Ser Leu Ser Ala Pro Gly Ser Glu Tyr 245 250 255 Gly Ser Pro Ser Val Ile Ser Val Ser Lys Gly Ser Pro Asp Gly Ser 260 265 270 His Pro Val Val Val Ala Pro Tyr Asn Gly Gly Pro Pro Arg Thr Cys 275 280 285 Pro Lys Ile Lys Gln Glu Ala Val Ser Ser Cys Thr His Leu Gly Ala 290 295 300 Gly Pro Pro Leu Ser Asn Gly His Arg Pro Ala Ala His Asp Phe Pro 305 310 315 320 Leu Gly Arg Gln Leu Pro Ser Arg Thr Thr Pro Thr Leu Gly Leu Glu 325 330 335 Glu Val Leu Ser Ser Arg Asp Cys His Pro Ala Leu Pro Leu Pro Pro 340 345 350 Gly Phe His Pro His Pro Gly Pro Asn Tyr Pro Ser Phe Leu Pro Asp 355 360 365 Gln Met Gln Pro Gln Val Pro Pro Leu His Tyr Gln Glu Leu Met Pro 370 375 380 Pro Gly Ser Cys Met Pro Glu Glu Pro Lys Pro Lys Arg Gly Arg Arg 385 390 395 400 Ser Trp Pro Arg Lys Arg Thr Ala Thr His Thr Cys Asp Tyr Ala Gly 405 410 415 Cys Gly Lys Thr Tyr Thr Lys Ser Ser His Leu Lys Ala His Leu Arg 420 425 430 Thr His Thr Gly Glu Lys Pro Tyr His Cys Asp Trp Asp Gly Cys Gly 435 440 445 Trp Lys Phe Ala Arg Ser Asp Glu Leu Thr Arg His Tyr Arg Lys His 450 455 460 Thr Gly His Arg Pro Phe Gln Cys Gln Lys Cys Asp Arg Ala Phe Ser 465 470 475 480 Arg Ser Asp His Leu Ala Leu His Met Lys Arg His Phe 485 490 <210> 247 <211> 1410 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-cMyc cDNA Sequence <400> 247 atgggttcgt cgcaccacca ccaccatcat aaaaaaaaac gcaaactgga cttcttccgt 60 gtggttgaaa accagcaacc gccggcgacc atgccgctga atgtgtcctt tacgaaccgc 120 aattatgatc tggactacga ttcagttcaa ccgtattttt actgcgacga agaagaaaac 180 ttctatcagc aacagcaaca gtccgaactg cagccgccgg caccgtcaga agatatctgg 240 aaaaaattcg aactgctgcc gaccccgccg ctgtcgccga gccgtcgctc cggtctgtgt 300 tctccgagtt acgtcgccgt gacgccgttt agtctgcgtg gtgacaatga tggcggtggc 360 ggttccttct caaccgcaga tcaactggaa atggttacgg aactgctggg cggtgacatg 420 gtcaaccaga gctttatctg cgatccggat gacgaaacct tcatcaaaaa catcatcatt 480 caggactgta tgtggtcggg ctttagcgcg gcggcaaaac tggtctctga aaaactggca 540 agttatcagg ctgcgcgtaa agattcgggt agcccgaacc cggcacgtgg tcattctgtt 600 tgcagtacca gctctctgta tctgcaggac ctgtcggccg cagctagcga atgtatcgat 660 ccgagcgtcg tgtttccgta cccgctgaat gatagttcct caccgaaatc ctgcgcatca 720 caagactcga gcgctttctc cccgtctagt gattcactgc tgtcctcaac cgaatcgagc 780 ccgcagggtt ctccggaacc gctggtgctg cacgaagaaa cgccgccgac cacgtctagt 840 gattctgaag aagaacaaga agacgaagaa gaaattgatg ttgtcagcgt tgaaaaacgt 900 caggcgccgg gcaaacgctc tgaaagtggt tccccgtcag ccggcggtca ttcgaaaccg 960 ccgcacagcc cgctggtgct gaaacgttgc catgtttcta cccatcagca caactatgcg 1020 gccccgccga gtacgcgtaa agactacccg gcagctaaac gcgtgaaact ggatagcgtt 1080 cgtgtcctgc gccagatttc taacaatcgt aaatgtacca gtccgcgctc ctcagatacg 1140 gaagaaaacg tcaaacgtcg cacccacaat gtgctggaac gccaacgtcg caatgaactg 1200 aaacgtagct ttttcgcact gcgcgatcag atcccggaac tggaaaacaa tgaaaaagct 1260 ccgaaagtgg ttatcctgaa aaaagcgacc gcctatattc tgtcggtgca agcggaagaa 1320 cagaaactga ttagcgaaga agatctgctg cgtaaacgtc gtgaacaact gaaacacaaa 1380 ctggaacaac tgcgtaattc ttgcgcttaa 1410 <210> 248 <211> 1440 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-145 MTD cMyc cDNA Sequence <400> 248 atgggttcgt cgcaccacca ccaccatcat aaaaaaaaac gcaaagcggc tgccccggtt 60 ctgctgctgc tgctgctgga cttcttccgt gtggttgaaa accagcaacc gccggcgacc 120 atgccgctga atgtgtcctt tacgaaccgc aattatgatc tggactacga ttcagttcaa 180 ccgtattttt actgcgacga agaagaaaac ttctatcagc aacagcaaca gtccgaactg 240 cagccgccgg caccgtcaga agatatctgg aaaaaattcg aactgctgcc gaccccgccg 300 ctgtcgccga gccgtcgctc cggtctgtgt tctccgagtt acgtcgccgt gacgccgttt 360 agtctgcgtg gtgacaatga tggcggtggc ggttccttct caaccgcaga tcaactggaa 420 atggttacgg aactgctggg cggtgacatg gtcaaccaga gctttatctg cgatccggat 480 gacgaaacct tcatcaaaaa catcatcatt caggactgta tgtggtcggg ctttagcgcg 540 gcggcaaaac tggtctctga aaaactggca agttatcagg ctgcgcgtaa agattcgggt 600 agcccgaacc cggcacgtgg tcattctgtt tgcagtacca gctctctgta tctgcaggac 660 ctgtcggccg cagctagcga atgtatcgat ccgagcgtcg tgtttccgta cccgctgaat 720 gatagttcct caccgaaatc ctgcgcatca caagactcga gcgctttctc cccgtctagt 780 gattcactgc tgtcctcaac cgaatcgagc ccgcagggtt ctccggaacc gctggtgctg 840 cacgaagaaa cgccgccgac cacgtctagt gattctgaag aagaacaaga agacgaagaa 900 gaaattgatg ttgtcagcgt tgaaaaacgt caggcgccgg gcaaacgctc tgaaagtggt 960 tccccgtcag ccggcggtca ttcgaaaccg ccgcacagcc cgctggtgct gaaacgttgc 1020 catgtttcta cccatcagca caactatgcg gccccgccga gtacgcgtaa agactacccg 1080 gcagctaaac gcgtgaaact ggatagcgtt cgtgtcctgc gccagatttc taacaatcgt 1140 aaatgtacca gtccgcgctc ctcagatacg gaagaaaacg tcaaacgtcg cacccacaat 1200 gtgctggaac gccaacgtcg caatgaactg aaacgtagct ttttcgcact gcgcgatcag 1260 atcccggaac tggaaaacaa tgaaaaagct ccgaaagtgg ttatcctgaa aaaagcgacc 1320 gcctatattc tgtcggtgca agcggaagaa cagaaactga ttagcgaaga agatctgctg 1380 cgtaaacgtc gtgaacaact gaaacacaaa ctggaacaac tgcgtaattc ttgcgcttaa 1440 1440 <210> 249 <211> 479 <212> PRT <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-145 MTD cMyc Amino Acid Sequence <400> 249 Met Gly Ser Ser His His His His His His Lys Lys Lys Arg Lys Ala 1 5 10 15 Ala Ala Pro Val Leu Leu Leu Leu Leu Leu Asp Phe Phe Arg Val Val 20 25 30 Glu Asn Gln Gln Pro Pro Ala Thr Met Pro Leu Asn Val Ser Phe Thr 35 40 45 Asn Arg Asn Tyr Asp Leu Asp Tyr Asp Ser Val Gln Pro Tyr Phe Tyr 50 55 60 Cys Asp Glu Glu Glu Asn Phe Tyr Gln Gln Gln Gln Gln Ser Glu Leu 65 70 75 80 Gln Pro Pro Ala Pro Ser Glu Asp Ile Trp Lys Lys Phe Glu Leu Leu 85 90 95 Pro Thr Pro Pro Leu Ser Pro Ser Arg Arg Ser Gly Leu Cys Ser Pro 100 105 110 Ser Tyr Val Ala Val Thr Pro Phe Ser Leu Arg Gly Asp Asn Asp Gly 115 120 125 Gly Gly Gly Ser Phe Ser Thr Ala Asp Gln Leu Glu Met Val Thr Glu 130 135 140 Leu Leu Gly Gly Asp Met Val Asn Gln Ser Phe Ile Cys Asp Pro Asp 145 150 155 160 Asp Glu Thr Phe Ile Lys Asn Ile Ile Ile Gln Asp Cys Met Trp Ser 165 170 175 Gly Phe Ser Ala Ala Ala Lys Leu Val Ser Glu Lys Leu Ala Ser Tyr 180 185 190 Gln Ala Ala Arg Lys Asp Ser Gly Ser Pro Asn Pro Ala Arg Gly His 195 200 205 Ser Val Cys Ser Thr Ser Ser Leu Tyr Leu Gln Asp Leu Ser Ala Ala 210 215 220 Ala Ser Glu Cys Ile Asp Pro Ser Val Val Phe Pro Tyr Pro Leu Asn 225 230 235 240 Asp Ser Ser Ser Pro Lys Ser Cys Ala Ser Gln Asp Ser Ser Ala Phe 245 250 255 Ser Pro Ser Ser Asp Ser Leu Leu Ser Ser Thr Glu Ser Ser Pro Gln 260 265 270 Gly Ser Pro Glu Pro Leu Val Leu His Glu Glu Thr Pro Pro Thr Thr 275 280 285 Ser Ser Asp Ser Glu Glu Glu Gln Glu Asp Glu Glu Glu Ile Asp Val 290 295 300 Val Ser Val Glu Lys Arg Gln Ala Pro Gly Lys Arg Ser Glu Ser Gly 305 310 315 320 Ser Pro Ser Ala Gly Gly His Ser Lys Pro Pro His Ser Pro Leu Val 325 330 335 Leu Lys Arg Cys His Val Ser Thr His Gln His Asn Tyr Ala Ala Pro 340 345 350 Pro Ser Thr Arg Lys Asp Tyr Pro Ala Ala Lys Arg Val Lys Leu Asp 355 360 365 Ser Val Arg Val Leu Arg Gln Ile Ser Asn Asn Arg Lys Cys Thr Ser 370 375 380 Pro Arg Ser Ser Asp Thr Glu Glu Asn Val Lys Arg Arg Thr His Asn 385 390 395 400 Val Leu Glu Arg Gln Arg Arg Asn Glu Leu Lys Arg Ser Phe Phe Ala 405 410 415 Leu Arg Asp Gln Ile Pro Glu Leu Glu Asn Asn Glu Lys Ala Pro Lys 420 425 430 Val Val Ile Leu Lys Lys Ala Thr Ala Tyr Ile Leu Ser Val Gln Ala 435 440 445 Glu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Leu Arg Lys Arg Arg 450 455 460 Glu Gln Leu Lys His Lys Leu Glu Gln Leu Arg Asn Ser Cys Ala 465 470 475 <210> 250 <211> 672 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-Lin28 cDNA Sequence <400> 250 atgggctcgt cgcatcatca tcatcatcat aaaaaaaaac gcaaaggttc ggttagcaac 60 cagcaatttg cgggcggttg cgccaaagcg gccgaagaag caccggaaga agctccggaa 120 gatgcagctc gtgcagcaga cgaaccgcag ctgctgcatg gcgcaggtat ttgtaaatgg 180 ttcaatgtcc gtatgggctt tggtttcctg tctatgaccg cacgtgctgg tgtggcactg 240 gatccgccgg tggacgtttt tgtccatcaa agcaaactgc acatggaagg cttccgttct 300 ctgaaagaag gtgaagctgt tgaatttacc ttcaaaaaat ctgccaaagg cctggaatcc 360 attcgcgtga cgggtccggg cggtgtgttt tgcatcggca gcgaacgtcg cccgaagggt 420 aaatcaatgc agaaacgtcg ctcgaaaggt gatcgctgct ataactgtgg cggtctggac 480 catcacgcca aagaatgcaa actgccgccg cagccgaaaa aatgccattt ctgtcaaagc 540 atctctcaca tggtcgcgag ttgtccgctg aaagcccagc aaggcccgtc cgcacagggt 600 aaaccgacgt actttcgtga agaagaagaa gaaatccact ccccgacgct gctgccggaa 660 gcccagaact aa 672 <210> 251 <211> 708 <212> DNA <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-132 MTD Lin28 cDNA Sequence <400> 251 atgggctcgt cgcatcatca tcatcatcat aaaaaaaaac gcaaagcagt cgtcgtcccg 60 gcaatcgtcc tggcggcacc gggttcggtt agcaaccagc aatttgcggg cggttgcgcc 120 aaagcggccg aagaagcacc ggaagaagct ccggaagatg cagctcgtgc agcagacgaa 180 ccgcagctgc tgcatggcgc aggtatttgt aaatggttca atgtccgtat gggctttggt 240 ttcctgtcta tgaccgcacg tgctggtgtg gcactggatc cgccggtgga cgtttttgtc 300 catcaaagca aactgcacat ggaaggcttc cgttctctga aagaaggtga agctgttgaa 360 tttaccttca aaaaatctgc caaaggcctg gaatccattc gcgtgacggg tccgggcggt 420 gtgttttgca tcggcagcga acgtcgcccg aagggtaaat caatgcagaa acgtcgctcg 480 aaaggtgatc gctgctataa ctgtggcggt ctggaccatc acgccaaaga atgcaaactg 540 ccgccgcagc cgaaaaaatg ccatttctgt caaagcatct ctcacatggt cgcgagttgt 600 ccgctgaaag cccagcaagg cccgtccgca cagggtaaac cgacgtactt tcgtgaagaa 660 gaagaagaaa tccactcccc gacgctgctg ccggaagccc agaactaa 708 <210> 252 <211> 235 <212> PRT <213> Artificial Sequence <220> <223> Codon optimized His-NLS-JO-132 MTD Lin28 Amino Acid Sequence <400> 252 Met Gly Ser Ser His His His His His His Lys Lys Lys Arg Lys Ala 1 5 10 15 Val Val Val Pro Ala Ile Val Leu Ala Ala Pro Gly Ser Val Ser Asn 20 25 30 Gln Gln Phe Ala Gly Gly Cys Ala Lys Ala Ala Glu Glu Ala Pro Glu 35 40 45 Glu Ala Pro Glu Asp Ala Ala Arg Ala Ala Asp Glu Pro Gln Leu Leu 50 55 60 His Gly Ala Gly Ile Cys Lys Trp Phe Asn Val Arg Met Gly Phe Gly 65 70 75 80 Phe Leu Ser Met Thr Ala Arg Ala Gly Val Ala Leu Asp Pro Pro Val 85 90 95 Asp Val Phe Val His Gln Ser Lys Leu His Met Glu Gly Phe Arg Ser 100 105 110 Leu Lys Glu Gly Glu Ala Val Glu Phe Thr Phe Lys Lys Ser Ala Lys 115 120 125 Gly Leu Glu Ser Ile Arg Val Thr Gly Pro Gly Gly Val Phe Cys Ile 130 135 140 Gly Ser Glu Arg Arg Pro Lys Gly Lys Ser Met Gln Lys Arg Arg Ser 145 150 155 160 Lys Gly Asp Arg Cys Tyr Asn Cys Gly Gly Leu Asp His His Ala Lys 165 170 175 Glu Cys Lys Leu Pro Pro Gln Pro Lys Lys Cys His Phe Cys Gln Ser 180 185 190 Ile Ser His Met Val Ala Ser Cys Pro Leu Lys Ala Gln Gln Gly Pro 195 200 205 Ser Ala Gln Gly Lys Pro Thr Tyr Phe Arg Glu Glu Glu Glu Glu Ile 210 215 220 His Ser Pro Thr Leu Leu Pro Glu Ala Gln Asn 225 230 235

Claims

서열번호: 2, 서열번호: 4, 서열번호: 6, 서열번호: 8, 서열번호: 10 및 서열번호: 12 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 역분화 전사인자 단백질의 한쪽 또는 양쪽 말단에 서열번호: 16 및 서열번호 18 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 거대분자 전달 도메인 (macromolecule transduction domain, MTD)이 융합되어 있는 세포투과성 역분화 전사인자 재조합 단백질.
제 1 항에 있어서,
상기 거대분자 전달 도메인이 서열번호: 16 의 아미노산 서열을 갖는 JO-84 MTD 및 서열번호: 18 의 아미노산 서열을 갖는 JO-86 MTD 중의 어느 하나인 것을 특징으로 하는 세포투과성 역분화 전사인자 재조합 단백질.
제 1 항에 있어서,
상기 재조합 단백질의 한쪽 말단에 세포핵 배치 서열 (nuclear localization sequence: NLS) 및 히스티딘-표지 (histidine-tag) 친화성 도메인으로부터 선택되는 1 종 이상이 융합되어 있는 것을 특징으로 하는 세포투과성 역분화 전사인자 재조합 단백질.
제 1 항 내지 제 3 항 중의 어느 한 항에 있어서,
상기 재조합 단백질이 하기로 구성된 군으로부터 선택되는 것을 특징으로 하는 세포투과성 역분화 전사인자 재조합 단백질:
서열번호: 2, 4, 6, 8, 10, 및 12 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 역분화 전사인자 N-말단에
서열번호: 16 의 아미노산 서열을 갖는 JO-84 MTD 가 융합된 재조합 단백질;
서열번호: 2, 4, 6, 8, 10, 및 12 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 역분화 전사인자 C-말단에
서열번호: 16 의 아미노산 서열을 갖는 JO-84 MTD 가 융합된 재조합 단백질;
서열번호: 2, 4, 6, 8, 10, 및 12 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 역분화 전사인자 양 말단에
서열번호: 16 의 아미노산 서열을 갖는 JO-84 MTD 가 융합된 재조합 단백질;
서열번호: 2, 4, 6, 8, 10, 및 12 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 역분화 전사인자 N-말단에
서열번호: 18 의 아미노산 서열을 갖는 JO-86 MTD 가 융합된 재조합 단백질;
서열번호: 2, 4, 6, 8, 10, 및 12 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 역분화 전사인자 C-말단에
서열번호: 18 의 아미노산 서열을 갖는 JO-86 MTD 가 융합된 재조합 단백질;
및
서열번호: 2, 4, 6, 8, 10, 및 12 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 역분화 전사인자 양 말단에
서열번호: 18 의 아미노산 서열을 갖는 JO-86 MTD 가 융합된 재조합 단백질.
제 1 항 내지 제 4 항 중의 어느 한 항에 있어서,
상기 역분화 전사인자 재조합 단백질이 Nanog 서열번호: 22, 24, 26, 28, 30 및 32, Oct4 서열번호: 50, 52, 54, 56, 58 및 60, Sox2 서열번호: 78, 80, 82, 84, 86 및 88, Klf4 서열번호: 106, 108, 110, 112, 114 및 116, cMyc 서열번호: 134, 136, 138, 140, 142 및 144, 및 Lin28 서열번호: 162, 164, 166, 168, 170 및 172 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 것을 특징으로 하는 세포투과성 역분화 전사인자 재조합 단백질.
제 1 항 내지 제 4 항 중의 어느 한 항에 있어서,
상기 역분화 전사인자 재조합 단백질이 Nanog 서열번호: 36, 38, 40, 42, 44 및 46, Oct4 서열번호: 64, 66, 68, 70, 72 및 74, Sox2 서열번호: 92, 94, 96, 98, 100 및 102, Klf4 서열번호: 120, 122, 124, 126, 128 및 130, cMyc 서열번호: 148, 150, 152, 154, 156 및 158, 및 Lin28 서열번호: 176, 178, 180, 182, 184 및 186 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 것을 특징으로 하는 세포투과성 역분화 전사인자 재조합 단백질.
제1항의 세포투과성 역분화 전사인자 재조합 단백질을 코딩하는 폴리뉴클레오티드.
제 7 항에 있어서,
상기 Nanog 폴리뉴클레오티드가 서열번호: 21, 23, 25, 27, 29 및 31, Oct4 서열번호: 49, 51, 53, 55, 57 및 59, Sox2 서열번호: 77, 79, 81, 83, 85 및 87, Klf4 서열번호: 105, 107, 109, 111, 113 및 115, cMyc 서열번호: 133, 135, 137, 139, 141 및 143, Lin28 서열번호: 161, 163, 165, 167, 169 및 171 의 염기서열로 구성된 군으로부터 선택되는 염기서열을 갖는 것을 특징으로 하는 폴리뉴클레오티드.
제 7 항에 있어서,
상기 폴리뉴클레오티드가 Nanog 서열번호: 35, 37,39, 41, 43 및 45, Oct4 서열번호: 63, 65, 67, 69, 71 및 73, Sox2 서열번호: 91, 93, 95, 97, 99 및 101, Klf4 서열번호: 119, 121, 123, 125, 127 및 129, cMyc 서열번호: 147, 149, 151, 153, 155 및 157, Lin28 서열번호: 175, 177, 179, 181, 183 및 185 의 염기서열로 구성된 군으로부터 선택되는 염기서열을 갖는 것을 특징으로 하는 폴리뉴클레오티드.
제 7 항의 폴리뉴클레오티드를 포함하는 재조합 발현벡터.
제 10 항에 있어서,
상기 재조합 발현벡터가 pET28a(+)-HNM₈₄Nanog, pET28a(+)-HNNanogM₈₄, pET28a(+)-HNM₈₄NanogM₈₄, pET28a(+)-HNM₈₆Nanog, pET28a(+)-HNNanogM₈₆, pET28a(+)-HNM₈₆NanogM₈₆, pET28a(+)-HNM₈₄Oct4, pET28a(+)-HNOct4M₈₄, pET28a(+)-HNM₈₄Oct4M₈₄, pET28a(+)-HNM₈₆Oct4, pET28a(+)-HNOct4M₈₆, pET28a(+)-HNM₈₆Oct4M₈₆, pET28a(+)-HNM₈₄Sox2, pET28a(+)-HNSox2M₈₄, pET28a(+)-HNM₈₄Sox2M₈₄, pET28a(+)-HNM₈₆Sox2, pET28a(+)-HNSox2M₈₆, pET28a(+)-HNM₈₆Sox2M₈₆, pET28a(+)-HNM₈₄Klf4, pET28a(+)-HNKlf4M₈₄, pET28a(+)-HNM₈₄Klf4M₈₄, pET28a(+)-HNM₈₆Klf4, pET28a(+)-HNKlf4M₈₆, pET28a(+)-HNM₈₆Klf4M₈₆, pET28a(+)-HNM₈₄cMyc, pET28a(+)-HNcMycM₈₄, pET28a(+)-HNM₈₄cMycM₈₄, pET28a(+)-HNM₈₆cMyc, pET28a(+)-HNcMycM₈₆, pET28a(+)-HNM₈₆cMycM₈₆, pET28a(+)-HNM₈₄Lin28, pET28a(+)-HNLin28M₈₄, pET28a(+)-HNM₈₄Lin28M₈₄, pET28a(+)-HNM₈₆Lin28, pET28a(+)-HNLin28M₈₆, 및 pET28a(+)-HNM₈₆Lin28M₈₆ 로 구성된 군으로부터 선택되는 것을 특징으로 하는 재조합 발현벡터.
제 10 항의 재조합 발현벡터로 형질전환된 형질전환 세균.
제 12 항에 있어서,
형질전환 세균이 대장균 DH5α/HNM₈₆Oct4 (기탁번호: KCTC 11640BP) 또는 DH5α/HNM₈₆cMyc (기탁번호: KCTC 11661BP)인 것을 특징으로 하는 형질전환 세균.
하기 단계를 포함하는 제1항의 세포투과성 역분화 전사인자 재조합 단백질을 생산하는 방법:
1) 제12항의 형질전환 세균을 배양하여 세포투과성 역분화 전사인자 재조합 단백질을 발현시키는 단계; 및
2) 발현된 세포투과성 역분화 전사인자 재조합 단백질을 배양액으로부터 회수하는 단계.
서열번호: 2 의 N 말단에 서열번호 224, 서열번호: 4 의 N 말단에 서열번호: 226, 서열번호: 6 의 N 말단에 서열번호: 234, 서열번호: 8 의 N 말단에 서열번호: 232, 서열번호: 10 의 N 말단에 서열번호: 230, 서열번호: 12 의 N 말단에 서열번호: 228 의 아미노산 서열을 갖는 거대분자 전달 도메인 (macromolecule transduction domain, MTD)이 융합되어 있는 세포투과성 역분화 전사인자 재조합 단백질.
제 15 항에 있어서,
상기 거대분자 전달 도메인이 서열번호: 224 의 아미노산 서열을 갖는 JO-10 MTD, 서열번호: 226 의 아미노산 서열을 갖는 JO-52 MTD, 서열번호: 228 의 아미노산 서열을 갖는 JO-132 MTD 및 서열번호: 230 의 아미노산 서열을 갖는 JO-145 MTD, 서열번호: 232 의 아미노산 서열을 갖는 JO-173 MTD, 서열번호: 234 의 아미노산 서열을 갖는 JO-181 MTD 의 어느 하나인 것을 특징으로 하는 세포투과성 역분화 전사인자 재조합 단백질.
제 15 항에 있어서,
상기 재조합 단백질의 한쪽 말단에 세포핵 배치 서열 (nuclear localization sequence: NLS) 및 히스티딘-표지 (histidine-tag) 친화성 도메인으로부터 선택되는 1 종 이상이 융합되어 있는 것을 특징으로 하는 세포투과성 역분화 전사인자 재조합 단백질.
제 15 항 내지 제 17 항 중의 어느 한 항에 있어서,
상기 역분화 전사인자 재조합 단백질이 서열번호: 237, 240, 243, 246, 249 및 252 의 아미노산 서열로 구성된 군으로부터 선택되는 아미노산 서열을 갖는 것을 특징으로 하는 세포투과성 역분화 전사인자 재조합 단백질.
제15항의 세포투과성 역분화 전사인자 재조합 단백질을 코딩하는 폴리뉴클레오티드.
제 19 항에 있어서,
상기 폴리뉴클레오티드가 서열번호: 236, 239, 242, 245, 248 및 251 의 염기서열로 구성된 군으로부터 선택되는 염기서열을 갖는 것을 특징으로 하는 폴리뉴클레오티드.
제 19 항의 폴리뉴클레오티드를 포함하는 재조합 발현벡터.
제 21 항에 있어서,
상기 재조합 발현벡터가 pET28a(+)-HNM₁₀Nanog, pET28a(+)-HNM₅₂Oct4, pET28a(+)-HNM₁₈₁Sox2, pET28a(+)-HNM₁₇₃Klf4, pET28a(+)-HNM₁₄₅cMyc, pET28a(+)-HNM₁₃₂Lin28 로 구성된 군으로부터 선택되는 것을 특징으로 하는 재조합 발현벡터.
제 21 항의 재조합 발현벡터로 형질전환된 형질전환 세균.
제 23 항에 있어서,
형질전환 세균이 대장균 DH5α/HNM₁₀Nanog (기탁번호: KCTC 11660BP), DH5α/HNM₁₈₁Sox2 (기탁번호: KCTC 11659BP), DH5α/HNM₁₇₃Klf4 (기탁번호: KCTC 11662BP), 또는 DH5α/HNM₁₃₂Lin28 (기탁번호: KCTC 11663BP)인 것을 특징으로 하는 형질전환 세균.
하기 단계를 포함하는 제15항의 세포투과성 역분화 전사인자 재조합 단백질을 생산하는 방법:
1) 제23항의 형질전환 세균을 배양하여 세포투과성 역분화 전사인자 재조합 단백질을 발현시키는 단계; 및
2) 발현된 세포투과성 역분화 전사인자 재조합 단백질을 배양액으로부터 회수하는 단계.