루젠에스씨아이의 유전자합성(Gene Synthesis) 서비스를 소개합니다.
Ø GC-rich는 물론 AT-rich, toxic sequences도
합성이 가능합니다.
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.
Ø 유전자 합성후 재조합 효소와 Sanger sequencing분석을
동시에 진행하여결과 파일을 유저분께 보내드립니다.
Ø 유전자 발현 및 정제 서비스는 물론 렌티바이러스 생성 및
형질전환 렌티바이러스 구축 서비스를
추가 진행 가능합니다.
Ø 유전자 합성서비스를 이용하신 유저분들에게
Standard Cloning Vector는 물론
Baculovirus/Insect, Mammalian, Yeast, Bacterial
발현벡터 클로닝 해드립니다.
Vector List
Standard Cloning Vector| Vector Name | Resistance | Promoter |
|---|
| pUC57 | AmpR | NA |
| pUC57-Kan | KanR | NA |
| pUC57-BsaI-Free | AmpR | NA |
| pUC19 | AmpR | NA |
| pUC18 | AmpR | NA |
| pUC57-1.8K | AmpR | NA |
| pUC57-Simple | AmpR | NA |
| pBlueScript II SK(+) | AmpR | T7/T3 |
Baculovirus/Insect ________________Expression________________ VectorsMammalian ________________Expression________________ VectorsYeast ________________Expression________________ Vectors| Vector Name | Resistance | Promoter | Special Instructions |
|---|
| pPIC 3.5k | AmpR | AOX1 | |
| pPIC9 | AmpR | AOX1 | N- α-factor secretion signal |
| pPIC9K | AmpR/KanR | AOX1 | N- α-factor secretion signal |
| pPICZA | Zeocin | AOX1 | C- Myc and 6*His |
| pPICZB | Zeocin | AOX1 | C- Myc and 6*His |
| pPICZC | Zeocin | AOX1 | C- Myc and 6*His |
| pPICZaA | Zeocin | AOX1 | N- α-factor secretion signal C- Myc and 6*His |
| pPICZalphaB | Zeocin | AOX1 | N- α-factor secretion signal C- Myc and 6*His |
| pPICZalphaC | Zeocin | AOX1 | N- α-factor secretion signal C- Myc and 6*His |
Bacterial ________________Expression________________ Vectors| Vector Name | Resistance | Promoter | Tags(N-Terminal) | Tags(C-Terminal) | Protease Cleavage Sites |
|---|
| pET-3a | AmpR | T7
| N- T7.Tag | | |
| pET-3d | AmpR | T7 | N- T7.Tag | | |
| pET-9a | KanR | T7
| N- T7.Tag | | |
| pET-11a | AmpR | T7 | N- T7.Tag | | |
| pET-11b | AmpR | T7
| N- T7.Tag | | |
| pET-11d | AmpR | T7 | N- T7.Tag | | |
| pET-15b | AmpR | T7
| N- His.Tag | | Thrombin site |
| pET-16b | AmpR | T7 | N- His.Tag | | Factor Xa site |
| pET-17b | AmpR | T7
| N- T7.Tag | | |
| pET-19b | AmpR | T7 | N- His.Tag | | Enterokinase site |
| pET-20b(+) | AmpR | T7
| N- His.Tag | C- Pelb signal | |
| pET-21a(+) | AmpR | T7 | N- T7.Tag | C- His.Tag | |
| pET-21b(+) | AmpR | T7
| N- T7.Tag | C- His.Tag | |
| pET-21d(+) | AmpR | T7 | N- T7.Tag | C- His.Tag | |
| pET-22b(+) | AmpR | T7
| N- Pelb signal | C- His.Tag | |
| pET-23a(+) | AmpR | T7 | N- T7.Tag | C- His.Tag | |
| pET-24a(+) | KanR | T7
| N- T7.Tag | C- His.Tag | |
| pET-24b(+) | KanR | T7 | N- T7.Tag | C- His.Tag | |
| pET-24c(+) | KanR | T7
| N- T7.Tag | C- His.Tag | |
| pET-24d(+) | KanR | T7 | N- T7.Tag | C- His.Tag | |
| pET-25b(+) | AmpR | T7
| N- Pelb signal | C- Hsv.Tag and His.Tag | |
| pET-26b(+) | KanR | T7 | N- Pelb signal | C- His.Tag | |
| pET-27b(+) | KanR | T7
| N- Pelb signal | C- Hsv.Tag and His.Tag | |
| pET-28a(+) | KanR | T7 | N- His.Tag and T7.Tag | C- His.Tag | Thrombin site |
| pET-28b(+) | KanR | T7
| N- His.Tag and T7.Tag | C- His.Tag | Thrombin site |
| pET-29a(+) | KanR | T7 | N- S.Tag | C- His.Tag | Thrombin site |
| pET-29b(+) | KanR | T7
| N- S.Tag | C- His.Tag | Thrombin site |
| pET-30a(+) | KanR | T7 | N- His.Tag and S.Tag | C- His.Tag | Thrombin site and Enterokinase site |
| pET-30b(+) | KanR | T7
| N- His.Tag and S.Tag | C- His.Tag | Thrombin site and Enterokinase site |
| pET-31b(+) | AmpR | T7 | N- KSI and His.Tag | C- His.Tag | |
| pET-32a(+) | AmpR | T7
| N- TrxA and His.Tag and S.Tag | C- His.Tag | Thrombin site and Enterokinase site |
| pET-32b(+) | AmpR | T7 | N- TrxA and His.Tag and S.Tag | C- His.Tag | Thrombin site and Enterokinase site |
| pET-41a(+) | KanR | T7
| N- GST.Tag and His.Tag and S.Tag | C- 8*His.Tag | Thrombin site and Enterokinase site |
| pET-41b(+) | KanR | T7 | N- GST.Tag and His.Tag and S.Tag | C- 8*His.Tag | Thrombin site and Enterokinase site |
| pET-42b(+) | KanR | T7
| N- GST.Tag and His.Tag and S.Tag | C- 8*His.Tag | Thrombin site and Factor Xa site |
| pET-43.1a(+) | AmpR | T7 | N- NusA and His.Tag and S.Tag | C- Hsv.Tag and His.Tag | Thrombin site |
| pET-45b(+) | AmpR | T7
| N- His.Tag | C- S.Tag | Enterokinase site |
| pET-51b(+) | AmpR | T7 | N- Strep-Tag II | C- 10*His.Tag | HRV 3C site |
| pET-52b(+) | AmpR | T7
| N- Strep-Tag II | C- 10*His.Tag | HRV 3C site |
| pGEX-4T-1 | AmpR | tac | N- GST.Tag | | Thrombin site |
| pGEX-4T-2 | AmpR | tac
| N- GST.Tag | | Thrombin site |
| pGEX-5X-1 | AmpR | tac | N- GST.Tag | | Factor Xa site |
| pGEX-6P-1 | AmpR | tac
| N- GST.Tag | | PreScission site |
| pGEX-6P-3 | AmpR | tac | N- GST.Tag | | PreScission site |
| pCDFDuet-1 | SmR | T7
| | C- S.Tag | |
| pMAL-c4x | AmpR | tac | N- MBP | | Factor Xa site |
| pMAL-c5E | AmpR | tac
| N- MBP | | Enterokinase site |
| pMAL-c5x | AmpR | tac | N- MBP | | Factor Xa site |
| pMAL-p5E | AmpR | tac
| N- MBP | | Enterokinase site |
| pMAl-p5x | AmpR | tac | N- MBP | | Factor Xa site |
| pQE-30 | AmpR | T5
| N- His.Tag | | |
| pQE-60 | AmpR | T5 | N- His.Tag | | |
| pGS-21a | AmpR | T7 | N- His.Tag and GST.Tag | | Enterokinase 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
