i90~210¹ Sizej
Code No. 160­25611 (20Êg)
5'­­TGGCAAAGAATTAACCCTCACTAAAGGGGTACCGGGCCCCCCCTCGAG
Kpn I
Xho I
GTCGAC GGTATCGATAAGCTTGATATCGAATTCCTGCAGCCCGGGGGATCC
Sal I/Acc I
EcoRV
BamH I
for Genetic Research
Wako
pCAG­Hyg
ACTAGTTCTAGAGCGGCCGCGACT­­3'
Spe I
Not I
pCAG is gene expression vector for mammalian cells.
pCAG includes CAG promoter in itself. CAG promoter is known to be
able to artificially induce high expression of interesting genes in mam­
malian cells.
œ
kMCS sequencing primersl
Forward primer : ttattgtgctgtctcatc
Reverse primer : GTGGTATGGCTGATTATG
kPurification methodl
Cesium chloride density­gradient centrifugation
kFeaturesl
œ
kMultiple cloning site (MCS)l
High expression by CAG promoter
Optimized expression level of drug resistance gene
kVector Mapl
10 mmol/L Tris­HCl (pH8.0), 1 mmol/L EDTA
kDNA Concentrationl
SR Promoter
1Êg/ÊL
Hygromycin BR
SV40 PolyA
kFormulationl
kSelection marker of E. coli and Mammalian cellsl
Hygromycin B phosphotransferase
MCS
pCAG-Hyg
kNuclease contamination checkl
5,925bp
Checked by electrophoresis
CAG Promoter
pUC ori
kHost mammalian cellsl
Primates (HeLa, HEK293, HepG2, COS­7, Vero, Jurkat, HL­60, etc),
Dog (MDCK), Rodents (NIH­3T3, FM3A, SCC131, PC12, CHO, etc).
kPropagation in E. coli l
DH1 E. coli strain (XL­1, XL­1 species, DH5¿), JM109 and XL10­Gold
is used for propagation of plasmid.
kSelection of E. coli transformant by Hygromycin Bl
Transformation of E. coli by heat shock methods
After heat shock procedures, pre­culture the E. coli in SOC liquid medium
at 37ú for 30|90 min.
kMCS promoterl
CAG
kReplication originl
E. coli : pUC ori
kVector elementsl
CAG promoter
MCS(Multiple cloning site)
SV40 early mRNA polyadenylation signal
1­1703
1708­1820
1826­2053
SR¿promoter for expression of Hygromycin B
phosphotransferase coding sequences
2668­3457
Hygromycin B phosphotransferase coding sequences
pUC plasmid replication origin
3596­4633
5259­5897
«
kStoragel
«
kPackagel
Spread pre­cultured E. coli onto LB agar medium including 50|60
Êg/mL of Hygromycin B.
|20ú
20Êg
Select some clones.
kTransfection into mammalian cellsl
We recommend you to use generally available transfection reagents. The
appropriate concentration of Hygromycin B for clone selection will be
optimized for your research cell lines.
kNotice to Purchaserl
Wako Products (the Products) are intended for research use only.
If you have required for sales and/or development of new vector which
were constructed by DNA fragments and/or sequence information of the
Products, please contact us. (labchem­tec—wako­chem.co.jp)
kVector sizel
Approx. 5.9kbp
| 1/6 |
| 2/6 |
R[h No. 160­25611 (20Êg)
â`q¤†p
pCAG­Hyg
pCAGÍA®¨×EÅgp·éâ`q­»xN^[Å·B
CAGv‚[^[Ì­«ÉæèA®¨×Eàɨ¢ÄAâ`q±üÉæ
éÚI^“pN¿Ì‚¢­»ªúÒÅ«Ü·B
kÁ
·l
œ CAGv‚[^[Éæ邭»U±
œ òÜÏ«â`qÌ­»ÊðÅK»
kxN^[}bvl
SR Promoter
Hygromycin BR
SV40 PolyA
MCS
pCAG-Hyg
5,925bp
CAG Promoter
pUC ori
kh宨×El
ì·Þ×E(HeLa, HEK293, HepG2, COS­7, Vero, Jurkat, HL­60ÈÇ)A
Ck×E(MDCK)A°Á•Þ×E(NIH­3T3, FM3A, SCC131, PC12,
CHOÈÇ)ÅgpÅ«Ü·B
kå°ÛÅ̝l
DH1nE. coli(XL­1AXL­1ޏ”ADH5¿)AJM109AܽÍXL10­
GoldÈÇå^xN^[¡»pÌå°Û”Ìgpð„§¢½µÜ·B
kå°Û`¿]·ãÌHygromycin BÉæéI²l
q[gV‡bN@Éæèå°ÛÌ`¿]·ðs¤ÛÉÍAq[gV‡b
NãÌå°ÛðSOCtÌ|nÅvŒJ‹`ƒ[(30`90ªA37ú)µA
50`60Êg/mL Hygromycin B ðÜÞLB¦V|nÉdíµÄ­¾³¢B
Wako Pure Chemical Industries, Ltd.
1­2, Doshomachi 3­Chome, Chuo­Ku, Osaka 540­8605, Japan
Telephone : {81­6­6203­3741
Facsimile : {81­6­6201­5964
http://www.wako­chem.co.jp
Wako Chemicals USA, Inc.
Wako Chemicals GmbH
1600 Bellwood Road
Richmond, VA 23237
U.S.A.
Telephone : {1­804­271­7677
Facsimile : {1­804­271­7791
http://www.wakousa.com
Fuggerstrasse 12
D­41468 Neuss
Germany
Telephone : {49­2131­311­0
Facsimile : {49­2131­311100
http://www.wako­chemicals.de
| 3/6 |
k®¨×EÅÌgpl
â`q±üÉÍêÊIÈg‰“XtFNV‡“Žòð²gpº³¢BÜ
½A²gpÌ×E”ÉæèAN[“I²pÌ Hygromycin B ÌZxª
ÏíèÜ·B{iÌgpOÉÍN[“I²Â\È Hygromycin B ÌI
Zxð 究߲Ÿ¢º³¢B
kxN^[TCYl
ñ5.9kbp
| 4/6 |
kMultiple cloning site (MCS)l
k‰CZ“XÉ¢Äl
5'­­TGGCAAAGAATTAACCCTCACTAAAGGGGTACCGGGCCCCCCCTCGAG
Kpn I
Xho I
GTCGAC GGTATCGATAAGCTTGATATCGAATTCCTGCAGCCCGGGGGATCC
Sal I/Acc I
EcoRV
BamH I
aõƒòHƔ®ïÐÌ»iiȺA{ijð²wüãÉͤ†prÅ
²gpº³¢B{iÌ]„A{iÉRˆ·éDNAfÐܽÍDNAzñ
îñðp¢ÄV½É\z³ê½v‰X~hÌ̄™ðÚIɲgp³ê
éê‡ÉÍA¾Ð¨qlŠkº(labchem­tec—wako­chem.co.jp)ÜŨ
ACTAGTTCTAGAGCGGCCGCGACT­­3'
Spe I
Not I
⇹º³¢B
kMCSV[NG“Xpv‰C}[l
Forward primer : ttattgtgctgtctcatc
Reverse primer : GTGGTATGGCTGATTATG
k¸ » @l
–»ZVE€§xùz“S@
kg
¬l
10 mmol/L Tris­HCl(pH 8.0), 1 mmol/L EDTA
kDNAZxl
1Êg/ÊL
kå°Û¥®¨×EI²}[J[l
Hygromycin B phosphotransferase
kNucleaseŸ`FbNl
dCj®ÅmFÏÝ
kMCSv‚[^[l
CAG
k¡»N_l
å°ÛFpUC ori
kxN^[zñTvl
CAG promoter
MCS(Multiple cloning site)
SV40 early mRNA polyadenylation signal
1­1703
1708­1820
1826­2053
SR¿promoter for expression of Hygromycin B
phosphotransferase coding sequences
2668­3457
Hygromycin B phosphotransferase coding sequences
pUC plasmid replication origin
3596­4633
5259­5897
kÛ¶ðl
|20ú
ke
Êl
STUVW XYZ[\
20Êg
1303K01
| 5/6 |
| 6/6 |

Class-X - angel Education Centre anand

pUC ori CAG Promoter MCS SV40 PolyA SRα Promoter BleomycinR