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질문 루젠SCI 유전자합성서비스는 어떻게 진행되나요?
작성자 lugensci
작성일자 2022-12-26

루젠에스씨아이의 유전자합성(Gene Synthesis) 서비스를 소개합니다.

 

GC-Rich 유전자도 합성이 가능한가요?

 

Ø  GC-rich는 물론 AT-rich, toxic sequences도

합성이 가능합니다.

 

유전자 길이는 얼마까지 합성이 가능한가요?

 

Ø  기본적으로 3Kb까지는 2주내에 합성완료되며 

6Kb이상까지도 검토후 합성진행 가능합니다.

 

단백질 서열정보만으로 유전자 합성이 가능한가요?

 

Ø  발현 호스트를 알려주시면 추가 비용 없이 코돈(codon) 최적화를 진행하여유전자를 합성해드립니다.

 

합성 유전자 정보는 어떻게 확인 가능한가요?

 

Ø  유전자 합성후 재조합 효소와 Sanger sequencing분석을

동시에 진행하여결과 파일을 유저분께 보내드립니다.

 

유전자 합성후 연계된 다른서비스가 있나요?

Ø  유전자 발현 및 정제 서비스는 물론 렌티바이러스 생성 및

형질전환 렌티바이러스 구축 서비스를 추가 진행 가능합니다.

 

발현 벡터에 클로닝이 가능한가요?

 

Ø  유전자 합성서비스를 이용하신 유저분들에게 

Standard Cloning Vector는 물론 

Baculovirus/Insect, Mammalian, Yeast, Bacterial


발현벡터 클로닝 해드립니다. 

Vector List

Standard Cloning Vector
Vector NameResistancePromoter
pUC57AmpRNA
pUC57-KanKanRNA
pUC57-BsaI-FreeAmpRNA
pUC19AmpRNA
pUC18AmpRNA
pUC57-1.8KAmpRNA
pUC57-SimpleAmpRNA
pBlueScript II SK(+)AmpRT7/T3

Baculovirus/Insect ________________Expression________________ Vectors
Vector NameResistancePromoter
pFastBac1AmpR/GmRpolyhedrin
pFastBac-DualAmpR/GmRp10 & polyhedrin

Mammalian ________________Expression________________ Vectors
Vector NameResistancePromoterSpecial Instructions
pcDNA3.1(+)NeoR/KanR/AmpRCMV /T7
pcDNA3.1(-)NeoR/KanR/AmpRCMV /T7
pcDNA3.1(+) myc-His ANeoR/KanR/AmpRCMV /T7C- Myc tag and 6xHis tag
pcDNA3.1(+) myc-His BNeoR/KanR/AmpRCMV /T7C- Myc tag and 6xHis tag
pcDNA3.1(+) myc-His CNeoR/KanR/AmpRCMV /T7C- Myc tag and 6xHis tag
pcDNA3.1 myc-His(-) ANeoR/KanR/AmpRCMV /T7C- Myc tag and 6xHis tag
pcDNA3.1 myc-His(-) CNeoR/KanR/AmpRCMV /T7C- Myc tag and 6xHis tag
pcDNA3.1(+)-EGFPNeoR/KanR/AmpRCMV /T7C- EGFP tag
pcDNA3.1/Hygro(+)HygR/AmpRCMV /T7
pcDNA3.1/Hygro(-)HygR/AmpRCMV /T7
pcDNA3.1/Zeo (+)BleoR/AmpRCMV /T7
pcDNA3.1/Zeo (-)BleoR/AmpRCMV /T7
psiCHECK2AmpRHSV TK /T7
pEGFP-N1NeoR/KanRCMVC- EGFP tag
pEGFP-C1NeoR/KanRCMVN- EGFP tag
pGL3-basicAmpRNAC- luciferase tag
pGL3-PromoterAmpRSV40C- luciferase tag

Yeast ________________Expression________________ Vectors
Vector NameResistancePromoterSpecial Instructions
pPIC 3.5kAmpRAOX1
pPIC9AmpRAOX1N- α-factor secretion signal
pPIC9KAmpR/KanRAOX1N- α-factor secretion signal
pPICZAZeocinAOX1C- Myc and 6*His
pPICZBZeocinAOX1C- Myc and 6*His
pPICZCZeocinAOX1C- Myc and 6*His
pPICZaAZeocinAOX1N- α-factor secretion signal C- Myc and 6*His
pPICZalphaBZeocinAOX1N- α-factor secretion signal C- Myc and 6*His
pPICZalphaCZeocinAOX1N- α-factor secretion signal C- Myc and 6*His

Bacterial ________________Expression________________ Vectors
Vector NameResistancePromoterTags(N-Terminal)Tags(C-Terminal)Protease Cleavage Sites
pET-3aAmpR

T7

N- T7.Tag
pET-3dAmpRT7N- T7.Tag
pET-9aKanR

T7

N- T7.Tag
pET-11aAmpRT7N- T7.Tag
pET-11bAmpR

T7

N- T7.Tag
pET-11dAmpRT7N- T7.Tag
pET-15bAmpR

T7

N- His.TagThrombin site
pET-16bAmpRT7N- His.TagFactor Xa site
pET-17bAmpR

T7

N- T7.Tag
pET-19bAmpRT7N- His.TagEnterokinase site
pET-20b(+)AmpR

T7

N- His.TagC- Pelb signal
pET-21a(+)AmpRT7N- T7.TagC- His.Tag
pET-21b(+)AmpR

T7

N- T7.TagC- His.Tag
pET-21d(+)AmpRT7N- T7.TagC- His.Tag
pET-22b(+)AmpR

T7

N- Pelb signalC- His.Tag
pET-23a(+)AmpRT7N- T7.TagC- His.Tag
pET-24a(+)KanR

T7

N- T7.TagC- His.Tag
pET-24b(+)KanRT7N- T7.TagC- His.Tag
pET-24c(+)KanR

T7

N- T7.TagC- His.Tag
pET-24d(+)KanRT7N- T7.TagC- His.Tag
pET-25b(+)AmpR

T7

N- Pelb signalC- Hsv.Tag and His.Tag
pET-26b(+)KanRT7N- Pelb signalC- His.Tag
pET-27b(+)KanR

T7

N- Pelb signalC- Hsv.Tag and His.Tag
pET-28a(+)KanRT7N- His.Tag and T7.TagC- His.TagThrombin site
pET-28b(+)KanR

T7

N- His.Tag and T7.TagC- His.TagThrombin site
pET-29a(+)KanRT7N- S.TagC- His.TagThrombin site
pET-29b(+)KanR

T7

N- S.TagC- His.TagThrombin site
pET-30a(+)KanRT7N- His.Tag and S.TagC- His.TagThrombin site and Enterokinase site
pET-30b(+)KanR

T7

N- His.Tag and S.TagC- His.TagThrombin site and Enterokinase site
pET-31b(+)AmpRT7N- KSI and His.TagC- His.Tag
pET-32a(+)AmpR

T7

N- TrxA and His.Tag and S.TagC- His.TagThrombin site and Enterokinase site
pET-32b(+)AmpRT7N- TrxA and His.Tag and S.TagC- His.TagThrombin site and Enterokinase site
pET-41a(+)KanR

T7

N- GST.Tag and His.Tag and S.TagC- 8*His.TagThrombin site and Enterokinase site
pET-41b(+)KanRT7N- GST.Tag and His.Tag and S.TagC- 8*His.TagThrombin site and Enterokinase site
pET-42b(+)KanR

T7

N- GST.Tag and His.Tag and S.TagC- 8*His.TagThrombin site and Factor Xa site
pET-43.1a(+)AmpRT7N- NusA and His.Tag and S.TagC- Hsv.Tag and His.TagThrombin site
pET-45b(+)AmpR

T7

N- His.TagC- S.TagEnterokinase site
pET-51b(+)AmpRT7N- Strep-Tag IIC- 10*His.TagHRV 3C site
pET-52b(+)AmpR

T7

N- Strep-Tag IIC- 10*His.TagHRV 3C site
pGEX-4T-1AmpRtacN- GST.TagThrombin site
pGEX-4T-2AmpR

tac

N- GST.TagThrombin site
pGEX-5X-1AmpRtacN- GST.TagFactor Xa site
pGEX-6P-1AmpR

tac

N- GST.TagPreScission site
pGEX-6P-3AmpRtacN- GST.TagPreScission site
pCDFDuet-1SmR

T7

C- S.Tag
pMAL-c4xAmpRtacN- MBPFactor Xa site
pMAL-c5EAmpR

tac

N- MBPEnterokinase site
pMAL-c5xAmpRtacN- MBPFactor Xa site
pMAL-p5EAmpR

tac

N- MBPEnterokinase site
pMAl-p5xAmpRtacN- MBPFactor Xa site
pQE-30AmpR

T5

N- His.Tag
pQE-60AmpRT5N- His.Tag
pGS-21aAmpRT7N- His.Tag and GST.TagEnterokinase site


1. Conversion of nucleic acids

(1) Moles and mass

1 ug 1,000bp DNA = 1.52 pmol
1 ug SV40 DNA (5,243bp) = 0.29 pmol
1 ug FX174 DNA (5,386bp) = 0.28 pmol
1 ug lambda phage DNA (48,502bp) = 0.03 pmol
1 ug M13mp18/19 DNA (7,249bp) = 0.21 pmol
1 ug pBR322 DNA (4,361bp) = 0.35 pmol
1 ug pUC18/19 DNA (2,686bp) = 0.57 pmol
1 pmol 1,000bp DNA = 0.66 ug
1 pmol M13mp18/19 DNA (7,249bp) = 4.78 ug
1 pmol SV40 DNA (5,243bp) = 3.46 ug
1 pmol pUC18/19 DNA (2,686bp) = 1.77 ug
1 pmol pBR322 DNA (4,361bp) = 2.88 ug
1 pmol lambda phage DNA (48,502bp) = 32.01 ug
1 pmol FX174 DNA (5,386bp) = 3.55 ug

(2) Absorbance and concentration

1 OD260 dsDNA = 50 mg/ml
1 OD260 ssDNA = 33 mg/ml
1 OD260 ssRNA = 40 mg/ml

(3) Molecular weight (MW)

Average MW of dsDNA base pair = 600 Daltons
Average MW of ssDNA base pair = 333 Daltons
Average molecular weight of RNA base is 340 Daltons

(4) Concentration of nucleic acid end

Linear DNA：pmol ends = pmol DNA´ (number of cuts ´2 + 2)
Circular DNA：pmol ends = pmol DNA´ number of cuts ´2
1 ug linear 1,000bp DNA = 3.04 pmol ends
1 ug linear FX174 DNA (5,386bp) = 0.56 pmol ends
1 ug linear pUC18/19 DNA (2,686bp) = 1.14 pmol ends
1 ug linear SV40 DNA (5,243bp) = 0.58 pmol ends
1 ug linear pBR322 DNA (4,361bp) = 0.7 pmol ends
1 ug linear lambda phage DNA (48,502bp) = 0.06 pmol ends
1 ug linear M13mp18/19 DNA (7,249bp) = 0.42 pmol ends

2. Conversion of nucleotides

(1) Molecular weight (MW)

MW (Da) = 333’N (number of bases)

(2) Concentration

C (mmol/L or pmol/ml) = OD260 / (0.01´N) C (ng/ml) = OD260’ MW / (0.01´N)
Tips: OD260—absorbance at 260 nm

(3) Double-stranded DNA with a melting point of oligonucleotides：

Shorter than 20bp double-stranded oligonucleotide: Tm = 2 (A + T) + 4 (G + C)
Longer than 20bp double-stranded oligonucleotide: Tm = 81.5 + 16.6 ( lg[J+] ) + 0.41 (%GC) (600/N)
Tips: N—Length of the primer (calculate the number of bases)；J+ —— Monovalent cation concentration

(4) Protein/DNA Conversions

1 kb DNA = 333 amino acid ≈ 3.7´104 Da
10,000 Da Protein ≈ 270 bp DNA
30,000 Da Protein ≈ 810 bp DNA
50,000 Da Protein ≈ 1350 bp DNA
100,000 Da Protein ≈ 2700 bp DNA
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