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Plasmid Files

phCMV2

Compact mammalian cell vector for high-level expression of N-terminally HA-tagged proteins.

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phCMV2 Sequence and MapphCMV2.dna
Map and Sequence File   
Sequence Author:  MoBiTec
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 AflIII - PciI (4203) ApaLI (3889) EcoO109I (3383) BsaI (3274) PfoI (3060) RsrII (2801) BsrDI (2518) PflFI - Tth111I (2403) FspI (2387) MscI (2367) PluTI (2288) SfoI (2286) NarI (2285) KasI (2284) BspDI * - ClaI * (2125) AseI (7) NdeI (234) SnaBI (340) BsmBI (589) BbsI (634) ATG BspEI (834) NheI (844) BmtI (848) BglII (855) PaeR7I - XhoI (859) HindIII (868) EcoRI (875) Acc65I (891) KpnI (895) PspOMI (899) TspMI - XmaI (902) ApaI (903) SmaI (904) BamHI (906) NotI (929) XbaI * (939) MfeI (1035) HpaI (1048) Bts α I (1124) AflII (1167) DraIII (1401) SexAI * (1874) SfiI (2060) BseRI (2103) StuI (2106) phCMV2 4261 bp
AflIII  (4203)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4203)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (3889)
1 site
G T G C A C C A C G T G
EcoO109I  (3383)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
BsaI  (3274)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
PfoI  (3060)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
RsrII  (2801)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BsrDI  (2518)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PflFI  (2403)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2403)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
FspI  (2387)
1 site
T G C G C A A C G C G T
MscI  (2367)
1 site
T G G C C A A C C G G T
PluTI  (2288)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (2286)
1 site
G G C G C C C C G C G G
NarI  (2285)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
KasI  (2284)
1 site
G G C G C C C C G C G G
BspDI  (2125)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2125)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
SnaBI  (340)
1 site
T A C G T A A T G C A T
BsmBI  (589)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BbsI  (634)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BspEI  (834)
1 site
T C C G G A A G G C C T
NheI  (844)
1 site
G C T A G C C G A T C G
BmtI  (848)
1 site
G C T A G C C G A T C G
BglII  (855)
1 site
A G A T C T T C T A G A
PaeR7I  (859)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (859)
1 site
C T C G A G G A G C T C
HindIII  (868)
1 site
A A G C T T T T C G A A
EcoRI  (875)
1 site
G A A T T C C T T A A G
Acc65I  (891)
1 site
G G T A C C C C A T G G
KpnI  (895)
1 site
G G T A C C C C A T G G
PspOMI  (899)
1 site
G G G C C C C C C G G G
TspMI  (902)
1 site
C C C G G G G G G C C C
XmaI  (902)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
ApaI  (903)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SmaI  (904)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (906)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
NotI  (929)
1 site
G C G G C C G C C G C C G G C G
XbaI  (939)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1035)
1 site
C A A T T G G T T A A C
HpaI  (1048)
1 site
G T T A A C C A A T T G
BtsαI  (1124)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
AflII  (1167)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
DraIII  (1401)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
SexAI  (1874)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (2060)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
BseRI  (2103)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
StuI  (2106)
1 site
A G G C C T T C C G G A
NeoR/KanR
2157 .. 2951  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
2157 .. 2951  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
ori
3559 .. 4147  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3559 .. 4147  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1177 .. 1632  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1177 .. 1632  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
1765 .. 2122  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1765 .. 2122  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
60 .. 364  =  305 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
60 .. 364  =  305 bp
human cytomegalovirus immediate early enhancer
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
1049 .. 1170  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1049 .. 1170  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1659 .. 1763  =  105 bp
AmpR promoter
1659 .. 1763  =  105 bp
CMV intron
704 .. 799  =  96 bp
modified intron from human cytomegalovirus (CMV)
CMV intron
704 .. 799  =  96 bp
modified intron from human cytomegalovirus (CMV)
MCS
855 .. 911  =  57 bp
multiple cloning site
MCS
855 .. 911  =  57 bp
multiple cloning site
HSV TK poly(A) signal
3183 .. 3230  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3183 .. 3230  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HA
820 .. 846  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
HA
820 .. 846  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
ATG
817 .. 819  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
817 .. 819  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 ori
1973 .. 2108  =  136 bp
SV40 origin of replication
SV40 ori
1973 .. 2108  =  136 bp
SV40 origin of replication
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