pCAGHyg PA tagN

i90~210¹ Sizej
Code No. 164­27711 (20Êg)
For genetic research
pCAG­Hyg PA tag­N
[Transfection into mammalian cells]
We recommend you to use general transfection reagents. (e.g., 293­73201
ScreenF ectTM A) The appropriate concentration of Hygromycin B for
clone selection will be optimized for your cell lines.
[Vector size]
Approx. 6.0kbp
pCAG is a gene expression vector for mammalian cells. pCAG vector
includes CAG promoter in itself. CAG promoter is known to be able to
artificially induce high expression of target recombinant proteins in
mammalian cells. pCAG­Hyg PA tag­N contains PA tag at N­terminal
part of multiple cloning site (MCS) and expresses PA­tagged proteins.
PA tag is consisted of 12 amino acids (GVAMPGAEDDVV) and has
been used as the tag for detection and/or purification of PA­tagged
recombinant proteins.
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164­27711 pCAG­Hyg PA tag­N
5’--TTATTGTGCTGTCTCATCATTTTGGCAAAGAATTAACCCTCACTAAAGG
KpnΣ
PA tag sequence
High expression by CAG promoter.
Optimized expression level of drug resistance gene.
PA tag is already inserted into N­terminal part of multiple cloning site
(MCS).
[Vector Map]
164­27711 pCAG­Hyg PA tag­N
XhoΣ
GGTACCATGGGCGTTGCCATGCCAGGTGCCGAAGATGATGTGGTGCTCGAG
SalΣ
EcoR Χ
ClaΣ
BamH Σ
GTCGACGGTATCGATAAGCTTGATATCGAATTCCTGCAGCCCGGGGGATCC
Spe Σ
[Features]
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[Multiple cloning site (MCS)]
Not Σ
ACTAGTTCTAGAGCGGCCGCGACTCTAGAT--3’
[MCS sequencing primers]
Forward primer : ttattgtgctgtctcatc
Reverse primer : GCAAGTAAAACCTCTACA
[Formulation]
10 mmol/L Tris­HCl, 1 mmol/L EDTA
[DNA Concentration]
1.0Êg/ÊL
[Selection marker of E. coli and Mammalian cells]
Hygromycin B phosphotransferase
[Nuclease contamination check]
Checked by electrophoresis
[MCS promoter]
CAG
[Replication origin]
[Host mammalian cells]
Primates (HeLa, HEK293, HepG2, COS­7, Vero, Jurkat, HL­60, etc),
Dog (MDCK), Rodents (NIH­3T3, FM3A, SCC131, PC12, CHO, etc).
[Replication in E. coli ]
DH1 E. coli strain (XL­1, XL­1 species, DH5¿), JM109 and XL10­Gold
are recommended for replication of plasmid.
[Selection of E. coli transformant by Hygromycin B]
Transformation to E. coli by heat shock methods
After heat shock procedures, pre­culture the E. coli in SOC liquid medium
at 37ú for 30­90 min.
«
Spread the pre­cultured E. coli onto LB agar medium including 50­60
Êg/mL of Hygromycin B.
E. coli : pUC ori
[Vector elements]
pCAG­Hyg PA tag
CAG promoter
PA tag
MCS (Multiple cloning site)
SV40 early mRNA polyadenylation signal
SR¿promoter for expression of Hygromycin B
phosphotransferase coding sequences
Hygromycin B phosphotransferase coding sequences
HSV Thymidine Kinase polyadenylation signal
pUC plasmid replication origin
«
Select some clones.
| 1/6 |
| 2/6 |
-N
1­1703
1738­1773
1774­1843
1856­2083
2698­3487
3626­4663
4657­4931
5290­5927
R[h No. 164­27711 (20Êg)
[Storage]
-20ú
â`q¤†p
[Package]
pCAG­Hyg PA tag­N
20Êg
[Notice to Purchaser]
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)
[Reference]
Fujii, Y. et al ., ``PA tag : A versatile protein tagging system using a super
high affinity antibody against a dodecapeptide derived from human
podoplanin.'', Protein Exp. Purif ., 95, 240­247 (2014).
Product Name
·B
pCAG­Hyg PA tag­NÍAPA tagðMultiple Cloning SiteiMCSjÌ
N –[¤ÉÂxN^[ÅAÚIÌg·¦^“pN¿ÉPA tagðtÁ³
¹Ä­»³¹Ü·BPA tagÍA12A~m_iGVAMPGAEDDVVjÅ
\¬³êĨèAPA tagZ‡g·¦^“pN¿ÌŸo⸻Égp³ê
ĢܷB
[Á
[Related products of PA tag series]
Code No.
pCAGÍA®¨×EÅgpÂ\Èâ`q­»pxN^[Å·BCAGv
‚[^[Ì­«ÉæèA®¨×EàÅÚI^“pN¿ð‚­»Å«Ü
Package
165­26901
pCAG­Hyg PA tag­C
20Êg
162­26911
pCAG­Hyg PA tag­N Signal Plus
20Êg
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CAGv‚[^[Éæ邭»U±
òÜÏ«â`qÌ­»ÊðÅK»
PA tag‹ÚxN^[
[xN^[}bv]
164­27711 pCAG­Hyg PA tag­N
[h宨×E]
ì·Þ×E
iHeLa, HEK293, HepG2, COS­7, Vero, Jurkat, HL­60ÈÇj
A
Ck×EiMDCKjA°Á•Þ×EiNIH­3T3, FM3A, SCC131, PC12,
CHOÈÇjÅgpÅ«Ü·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 |
[å°ÛÅ̝]
DH1 n E. coli i XL­1 A XL­1 ޏ”A DH5¿jA JM109 AܽÍ
XL10­Gold ÈÇå^xN^[¡»pÌå°Û”Ìgpð„§¢½µÜ
·B
[å°Û`¿]·ãÌ Hygromycin BÉ æéI²]
q[gV‡bN@Éæèå°ÛÌ`¿]·ðs¤ÛÉÍAq[gV‡b
NãÌå°ÛðSOCtÌ|nÅvŒJ‹`ƒ[i30­90ªA37újµA
50­60Êg/mL Hygromycin BðÜÞ LB ¦V|nÉdíµÄ­¾³¢B
| 4/6 |
[®¨×EÅÌgp]
[Û¶ð]
â`q±üÉÍêÊIÈg‰“XtFNV‡“Žòð²gpº³¢B
iáF293­73201 ScreenF ectTM AjܽA²gpÌ×E”ÉæèAN[
|20ú
“I²pÌHygromycin BÌZxªÏíèÜ·B
{iÌgpOÉÍN[
“I²Â\ÈHygromycin BÌIZxð 究߲Ÿ¢º³¢B
[e
[xN^[TCY]
[‰CZ“XÉ¢Ä]
ñ 6.0kbp
Ê]
20Êg
aõƒòHƔ®ïÐÌ»iiȺA{ijð²wüãÉͤ†prÅ
²gpº³¢B{iÌ]„A{iÉRˆ·éDNAfÐܽÍDNAzñ
[Multiple cloning siteiMCSj]
îñðp¢ÄV½É\z³ê½v‰X~hÌ̄™ðÚIɲgp³ê
164­27711 pCAG­Hyg PA tag­N
éê‡ÉÍA¾Ð¨qlŠkºilabchem­tec—wako­chem.co.jpjÜÅ
¨â‡¹º³¢B
5’--TTATTGTGCTGTCTCATCATTTTGGCAAAGAATTAACCCTCACTAAAGG
PA tag ㈩೉
KpnΣ
XhoΣ
GGTACCATGGGCGTTGCCATGCCAGGTGCCGAAGATGATGTGGTGCTCGAG
EcoR Χ
ClaΣ
SalΣ
BamH Σ
GTCGACGGTATCGATAAGCTTGATATCGAATTCCTGCAGCCCGGGGGATCC
Spe Σ
high affinity antibody against a dodecapeptide derived from human
podoplanin.'', Protein Exp. Purif ., 95, 240­247 (2014).
Not Σ
ACTAGTTCTAGAGCGGCCGCGACTCTAGAT--3’
[PA tagVŠ[YÖA»i]
R[h No.
[MCSV[NG“Xpv‰C}[]
Forward primer : ttattgtgctgtctcatc
Reverse primer : GCAAGTAAAACCTCTACA
[g
[Ql¶£]
Fujii, Y. et al ., ``PA tag : A versatile protein tagging system using a super
i
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e
Ê
165­26901
pCAG­Hyg PA tag­C
20Êg
162­26911
pCAG­Hyg PA tag­N Signal Plus
20Êg
¬]
10 mmol/L Tris­HCl, 1 mmol/L EDTA
[DNAZx]
1.0Êg/ÊL
[å°Û¥®¨×EI²}[J[]
Hygromycin B phosphotransferase
[NucleaseŸ`FbN]
dCj®ÅmFÏÝ
[MCSv‚[^[]
CAG
[¡»N_]
å°Û F pUC ori
[xN^[zñTv]
pCAG­Hyg PA tag
CAG promoter
PA tag
MCS (Multiple cloning site)
SV40 early mRNA polyadenylation signal
SR¿promoter for expression of Hygromycin B
phosphotransferase coding sequences
Hygromycin B phosphotransferase coding sequences
HSV Thymidine Kinase polyadenylation signal
pUC plasmid replication origin
| 5/6 |
-N
1­1703
1738­1773
1774­1843
1856­2083
2698­3487
3626­4663
4657­4931
5290­5927
STUVW XYZ[\
1606K01
| 6/6 |