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Plasmid Files

pCMV-HA-N

Vector for expressing an N-terminally HA-tagged protein in mammalian cells. Identical to pCMV‑HA.

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pCMV-HA-N Sequence and MappCMV-HA-N.dna
Map and Sequence File   
Sequence Author:  Clontech
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 PluTI (3626) SfoI (3624) NarI (3623) KasI (3622) BstAPI (3572) SspI (3204) XmnI (2999) ScaI (2880) NmeAIII (2548) BsrFI (2480) BsaI (2461) AhdI (2400) AlwNI (1923) BfuAI - BspMI (12) SbfI (23) CMV enhancer BtgZI (300) BsaAI - SnaBI (306) Eco53kI (532) SacI (534) BbsI (600) BspEI (617) PpuMI (702) BseRI (730) StuI (769) PspOMI (820) ApaI (824) ATG MscI (865) SfiI (871) EcoRI (878) SalI (885) AccI (886) BglII (893) AvaI - BsoBI - PaeR7I - XhoI (899) BmeT110I (900) Acc65I (904) KpnI (908) EagI - NotI (911) PsiI (1035) MfeI (1064) NheI (1146) BmtI (1150) M13 rev lac operator lac promoter BspQI - SapI (1391) AflIII - PciI (1507) BseYI (1811) PspFI (1815) pCMV-HA-N 3782 bp
PluTI  (3626)
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  (3624)
1 site
G G C G C C C C G C G G
NarI  (3623)
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  (3622)
1 site
G G C G C C C C G C G G
BstAPI  (3572)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
SspI  (3204)
1 site
A A T A T T T T A T A A
XmnI  (2999)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2880)
1 site
A G T A C T T C A T G A
NmeAIII  (2548)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsrFI  (2480)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
BsaI  (2461)
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.
AhdI  (2400)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
AlwNI  (1923)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
BfuAI  (12)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (12)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
SbfI  (23)
1 site
C C T G C A G G G G A C G T C C
BtgZI  (300)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BsaAI  (306)
1 site
Y A C G T R R T G C A Y
SnaBI  (306)
1 site
T A C G T A A T G C A T
Eco53kI  (532)
1 site
G A G C T C C T C G A G
SacI  (534)
1 site
G A G C T C C T C G A G
BbsI  (600)
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  (617)
1 site
T C C G G A A G G C C T
PpuMI  (702)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BseRI  (730)
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  (769)
1 site
A G G C C T T C C G G A
PspOMI  (820)
1 site
G G G C C C C C C G G G
ApaI  (824)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
MscI  (865)
1 site
T G G C C A A C C G G T
SfiI  (871)
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.
EcoRI  (878)
1 site
G A A T T C C T T A A G
SalI  (885)
1 site
G T C G A C C A G C T G
AccI  (886)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
BglII  (893)
1 site
A G A T C T T C T A G A
AvaI  (899)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (899)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
PaeR7I  (899)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (899)
1 site
C T C G A G G A G C T C
BmeT110I  (900)
1 site
C Y C G R G G R G C Y C
Acc65I  (904)
1 site
G G T A C C C C A T G G
KpnI  (908)
1 site
G G T A C C C C A T G G
EagI  (911)
1 site
C G G C C G G C C G G C
NotI  (911)
1 site
G C G G C C G C C G C C G G C G
PsiI  (1035)
1 site
T T A T A A A A T A T T
MfeI  (1064)
1 site
C A A T T G G T T A A C
NheI  (1146)
1 site
G C T A G C C G A T C G
BmtI  (1150)
1 site
G C T A G C C G A T C G
BspQI  (1391)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (1391)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
AflIII  (1507)
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  (1507)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (1811)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
PspFI  (1815)
1 site
C C C A G C G G G T C G
AmpR
2327 .. 3187  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2327 .. 3118  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2327 .. 3187  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3119 .. 3187  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2327 .. 3187  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1568 .. 2156  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1568 .. 2156  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
27 .. 330  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
27 .. 330  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
331 .. 534  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
331 .. 534  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
921 .. 1055  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
921 .. 1055  =  135 bp
SV40 polyadenylation signal
AmpR promoter
3188 .. 3292  =  105 bp
AmpR promoter
3188 .. 3292  =  105 bp
SV40 intron
672 .. 768  =  97 bp
modified SV40 intron with splice donor and acceptor
sites
SV40 intron
672 .. 768  =  97 bp
modified SV40 intron with splice donor and acceptor
sites
MCS
863 .. 917  =  55 bp
multiple cloning site
MCS
863 .. 917  =  55 bp
multiple cloning site
lac promoter
1214 .. 1244  =  31 bp
   Segment 3:  -10  
   1214 .. 1220  =  7 bp
promoter for the E. coli lac operon
lac promoter
1214 .. 1244  =  31 bp
   Segment 2:  
   1221 .. 1238  =  18 bp
promoter for the E. coli lac operon
lac promoter
1214 .. 1244  =  31 bp
   Segment 1:  -35  
   1239 .. 1244  =  6 bp
promoter for the E. coli lac operon
lac promoter
1214 .. 1244  =  31 bp
3 segments
promoter for the E. coli lac operon
HA
832 .. 858  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
HA
832 .. 858  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin)
epitope tag
M13 rev
1166 .. 1182  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
1166 .. 1182  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
1190 .. 1206  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
1190 .. 1206  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
M13 fwd
3766 .. 3782  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
3766 .. 3782  =  17 bp
common sequencing primer, one of multiple similar
variants
ATG
829 .. 831  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
829 .. 831  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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