Resources
Plasmid Files

pCMV SPORT4

Vector for cloning and transient mammalian cell expression of cDNAs.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pCMV SPORT4.dna
Map and Sequence File:    Download    Open   
Sequence Author:  I.M.A.G.E. Consortium
Download Free Trial Get SnapGene Viewer


NaeI (4258) NgoMIV (4256) DraIII (4155) PflMI * (3843) XmnI (3455) ScaI (3336) BsaI (2917) AhdI (2856) AlwNI (2337) PspFI (2229) BseYI (2225) BspDI * - ClaI * (123) HpaI (259) MfeI (268) BsaBI * (360) BclI * (361) NheI (637) BmtI (641) SphI (731) MluI (733) HindIII (739) PspOMI (745) ApaI (749) AbsI - PaeR7I - PspXI - XhoI (751) BamHI (757) XbaI (763) EagI - NotI (770) SpeI (779) SalI (791) AccI (792) TspMI - XmaI (796) SmaI (798) EcoRI (801) RsrII (808) AgeI (810) Acc65I (813) KpnI (817) PstI - SbfI (822) BfuAI - BspMI (825) StuI (865) SacII (912) AvrII (915) BbsI (944) BsmBI (989) NcoI (1218) SnaBI (1242) NsiI (1529) BstAPI (1745) BspQI - SapI (1805) PciI (1921) pCMV SPORT4 4387 bp
NaeI  (4258)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (4256)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
DraIII  (4155)
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.
PflMI  (3843)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
XmnI  (3455)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3336)
1 site
A G T A C T T C A T G A
BsaI  (2917)
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  (2856)
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  (2337)
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.
PspFI  (2229)
1 site
C C C A G C G G G T C G
BseYI  (2225)
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.
BspDI  (123)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (123)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
HpaI  (259)
1 site
G T T A A C C A A T T G
MfeI  (268)
1 site
C A A T T G G T T A A C
BsaBI  (360)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BclI  (361)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
NheI  (637)
1 site
G C T A G C C G A T C G
BmtI  (641)
1 site
G C T A G C C G A T C G
SphI  (731)
1 site
G C A T G C C G T A C G
MluI  (733)
1 site
A C G C G T T G C G C A
HindIII  (739)
1 site
A A G C T T T T C G A A
PspOMI  (745)
1 site
G G G C C C C C C G G G
ApaI  (749)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AbsI  (751)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (751)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (751)
1 site
V C T C G A G B B G A G C T C V
XhoI  (751)
1 site
C T C G A G G A G C T C
BamHI  (757)
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.
XbaI  (763)
1 site
T C T A G A A G A T C T
EagI  (770)
1 site
C G G C C G G C C G G C
NotI  (770)
1 site
G C G G C C G C C G C C G G C G
SpeI  (779)
1 site
A C T A G T T G A T C A
SalI  (791)
1 site
G T C G A C C A G C T G
AccI  (792)
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.
TspMI  (796)
1 site
C C C G G G G G G C C C
XmaI  (796)
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.
SmaI  (798)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (801)
1 site
G A A T T C C T T A A G
RsrII  (808)
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.
AgeI  (810)
1 site
A C C G G T T G G C C A
Acc65I  (813)
1 site
G G T A C C C C A T G G
KpnI  (817)
1 site
G G T A C C C C A T G G
PstI  (822)
1 site
C T G C A G G A C G T C
SbfI  (822)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (825)
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  (825)
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.
StuI  (865)
1 site
A G G C C T T C C G G A
SacII  (912)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
AvrII  (915)
1 site
C C T A G G G G A T C C
BbsI  (944)
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.
BsmBI  (989)
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.
NcoI  (1218)
1 site
C C A T G G G G T A C C
SnaBI  (1242)
1 site
T A C G T A A T G C A T
NsiI  (1529)
1 site
A T G C A T T A C G T A
BstAPI  (1745)
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.
BspQI  (1805)
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  (1805)
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.
PciI  (1921)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
2783 .. 3643  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2783 .. 3574  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2783 .. 3643  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3575 .. 3643  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2783 .. 3643  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1982 .. 2570  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1982 .. 2570  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3931 .. 4386  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3931 .. 4386  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
1219 .. 1522  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1219 .. 1522  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
1015 .. 1218  =  204 bp
human cytomegalovirus (CMV) immediate early promoter