i90~210¹ Sizej
Code No. 163­25601 (20Êg)
5'­­TGGCAAAGAATTAACCCTCACTAAAGGGGTACCGGGCCCCCCCTCGAG
Kpn I
Xho I
GTCGAC GGTATCGATAAGCTT GATATCGAATTC CTGCAG CCCGGG GGAT
EcoRV
Sal I/Acc I
EcoRI
Pst I
Sma I BamHI
for Genetic Research
Wako
pCAG­Neo
CC 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
SV40 Promoter
NeomycinR
/KanamycinR
SV40 PolyA
kFormulationl
10 mmol/L Tris­HCl (pH8.0), 1 mmol/L EDTA
kDNA Concentrationl
1Êg/ÊL
kSelection marker of E. coli and Mammalian cellsl
Neomycin phosphotransferase (E. coli : Kanamycin, Mammalian cells :
G418)
MCS
pCAG-Neo
5,145bp
kNuclease contamination checkl
Checked by electrophoresis
CAG Promoter
pUC ori
kMCS promoterl
CAG
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 Kanamycinl
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.
«
kReplication originl
E. coli : pUC ori
kVector elementsl
CAG promoter
MCS(Multiple cloning site)
SV40 early mRNA polyadenylation signal
1­1703
1708­1820
1812­2053
SV40 promoter for expression of Neomycin
phosphotransferase coding sequences
2812­3004
Neomycin phosphotransferase coding sequences
pUC plasmid replication origin
3041­3835
4480­5117
Spread pre­cultured E. coli onto LB agar medium including 25|50
Êg/mL of Kanamycin.
kStoragel
Select some clones.
kPackagel
«
|20ú
20Êg
kTransfection into mammalian cellsl
We recommend you to use generally available transfection reagents. The
appropriate concentration of G418 for clone selection will be optimized
for your research cell lines.
kVector sizel
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)
Approx. 5.1kbp
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R[h No. 163­25601 (20Êg)
â`q¤†p
pCAG­Neo
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
SV40 Promoter
NeomycinR
/KanamycinR
SV40 PolyA
MCS
pCAG-Neo
5,145bp
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`¿]·ãÌKanamycinÉæéI²l
q[gV‡bN@Éæèå°ÛÌ`¿]·ðs¤ÛÉÍAq[gV‡b
NãÌå°ÛðSOCtÌ|nÅvŒJ‹`ƒ[(30`90ªA37ú)µA
25|50Êg/mL KanamycinðÜÞLB¦V|nÉdíµÄ­¾³¢B
Wako Pure Chemical Industries, Ltd.
k®¨×EÅÌgpl
â`q±üÉÍêÊIÈg‰“XtFNV‡“Žòð²gpº³¢BÜ
1­2, Doshomachi 3­Chome, Chuo­Ku, Osaka 540­8605, Japan
Telephone : {81­6­6203­3741
Facsimile : {81­6­6201­5964
http://www.wako­chem.co.jp
½A²gpÌ×E”ÉæèAN[“I²pÌG418ÌZxªÏíèÜ
Wako Chemicals USA, Inc.
Wako Chemicals GmbH
²Ÿ¢º³¢B
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
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·B{iÌgpOÉÍN[“I²Â\ÈG418ÌIZxð 究ß
kxN^[TCYl
ñ5.1kbp
| 4/6 |
kMultiple cloning site (MCS)l
k‰CZ“XÉ¢Äl
5'­­TGGCAAAGAATTAACCCTCACTAAAGGGGTACCGGGCCCCCCCTCGAG
Kpn I
Xho I
GTCGAC GGTATCGATAAGCTT GATATCGAATTC CTGCAG CCCGGG GGAT
EcoRV
Sal I/Acc I
EcoRI
Pst I
Sma I BamHI
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
CC 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
Neomycin phosphotransferase(E. coli : Kanamycin, Mammalian cells :
G418j
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
1812­2053
SV40 promoter for expression of Neomycin
phosphotransferase coding sequences
2812­3004
Neomycin phosphotransferase coding sequences
pUC plasmid replication origin
3041­3835
4480­5117
kÛ¶ðl
|20ú
ke
STUVW XYZ[\
Êl
20Êg
1303K01
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( mercury )