pCMV SPORT

Vector for cloning and transient mammalian cell expression of cDNAs.

Sequence Author: I.M.A.G.E. Consortium

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BspDI * - ClaI * (123) NaeI (4251) NgoMIV (4249) DraIII (4148) PflMI * (3836) XmnI (3448) ScaI (3329) BsaI (2910) AhdI (2849) AlwNI (2330) PspFI (2222) BseYI (2218) HpaI (259) MfeI (268) BsaBI * (360) BclI * (361) NheI (637) BmtI (641) T7 promoter SphI (686) MluI (688) HindIII (694) PspOMI (700) ApaI (704) AbsI - PaeR7I - PspXI - XhoI (706) BamHI (712) XbaI (718) EagI - NotI (725) SpeI (734) SalI (746) TspMI - XmaI (751) SmaI (753) EcoRI (756) RsrII (763) AgeI (765) Acc65I (768) KpnI (772) PstI - SbfI (777) SP6 promoter AvrII (821) BbsI (850) BsmBI - Esp3I (895) BtgI - NcoI (1124) SnaBI (1148) BsgI * (1416) lac operator lac UV5 promoter BstZ17I (1547) BstAPI (1738) BspQI - SapI (1798) PciI (1914) pCMV SPORT 4380 bp
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.
NaeI  (4251)
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  (4249)
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  (4148)
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  (3836)
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  (3448)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3329)
1 site
A G T A C T T C A T G A
BsaI  (2910)
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  (2849)
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  (2330)
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  (2222)
1 site
C C C A G C G G G T C G
BseYI  (2218)
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.
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  (686)
1 site
G C A T G C C G T A C G
MluI  (688)
1 site
A C G C G T T G C G C A
HindIII  (694)
1 site
A A G C T T T T C G A A
PspOMI  (700)
1 site
G G G C C C C C C G G G
ApaI  (704)
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  (706)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (706)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (706)
1 site
V C T C G A G B B G A G C T C V
XhoI  (706)
1 site
C T C G A G G A G C T C
BamHI  (712)
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  (718)
1 site
T C T A G A A G A T C T
EagI  (725)
1 site
C G G C C G G C C G G C
NotI  (725)
1 site
G C G G C C G C C G C C G G C G
SpeI  (734)
1 site
A C T A G T T G A T C A
SalI  (746)
1 site
G T C G A C C A G C T G
TspMI  (751)
1 site
C C C G G G G G G C C C
XmaI  (751)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (753)
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  (756)
1 site
G A A T T C C T T A A G
RsrII  (763)
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  (765)
1 site
A C C G G T T G G C C A
Acc65I  (768)
1 site
G G T A C C C C A T G G
KpnI  (772)
1 site
G G T A C C C C A T G G
PstI  (777)
1 site
C T G C A G G A C G T C
SbfI  (777)
1 site
C C T G C A G G G G A C G T C C
AvrII  (821)
1 site
C C T A G G G G A T C C
BbsI  (850)
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  (895)
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (895)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BtgI  (1124)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1124)
1 site
C C A T G G G G T A C C
SnaBI  (1148)
1 site
T A C G T A A T G C A T
BsgI  (1416)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
* Blocked by EcoKI methylation.
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BstZ17I  (1547)
1 site
G T A T A C C A T A T G
BstAPI  (1738)
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  (1798)
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  (1798)
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  (1914)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
2776 .. 3636  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2776 .. 3567  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2776 .. 3636  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3568 .. 3636  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2776 .. 3636  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1975 .. 2563  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1975 .. 2563  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3924 .. 4379  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3924 .. 4379  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
1125 .. 1428  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1125 .. 1428  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
921 .. 1124  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
921 .. 1124  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
125 .. 259  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
125 .. 259  =  135 bp
SV40 polyadenylation signal
MCS
682 .. 779  =  98 bp
multiple cloning site
MCS
682 .. 779  =  98 bp
multiple cloning site
loxP
2641 .. 2674  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
2641 .. 2674  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
lac UV5 promoter
1482 .. 1513  =  32 bp
3 segments
   Segment 3:  -10  
   1482 .. 1488  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1482 .. 1513  =  32 bp
3 segments
   Segment 2:  
   1489 .. 1507  =  19 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1482 .. 1513  =  32 bp
3 segments
   Segment 1:  -35  
   1508 .. 1513  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1482 .. 1513  =  32 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
646 .. 664  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
646 .. 664  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
787 .. 805  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
787 .. 805  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
lac operator
1458 .. 1474  =  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
1458 .. 1474  =  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).
ORF:  3622 .. 3861  =  240 bp
ORF:  79 amino acids  =  9.0 kDa
ORF:  2906 .. 3172  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2776 .. 3636  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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