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Plasmid Files

pCMV SPORT2

Vector for cloning and transient mammalian cell expression of cDNAs.

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pCMV SPORT2 Sequence and MappCMV SPORT2.dna
Map and Sequence File   
Sequence Author:  I.M.A.G.E. Consortium
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 BspDI * - ClaI * (123) NaeI (4345) NgoMIV (4343) DraIII (4242) PflMI * (3930) XmnI (3542) ScaI (3423) BsaI (3004) AhdI (2943) AlwNI (2424) PspFI (2316) BseYI (2312) HpaI (259) MfeI (268) BsaBI * (360) BclI * (361) NheI (637) BmtI (641) BglII (643) T7 promoter 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) TspMI - XmaI (796) SmaI (798) EcoRI (801) RsrII (808) AgeI (810) Acc65I (813) KpnI (817) PstI - SbfI (822) SP6 promoter M13 rev SacII (912) AvrII (915) BbsI (944) BsmBI (989) NcoI (1218) SnaBI (1242) BsgI * (1510) lac operator lac UV5 promoter BstZ17I (1641) BstAPI (1832) BspQI - SapI (1892) PciI (2008) pCMV SPORT2 4474 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  (4345)
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  (4343)
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  (4242)
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  (3930)
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  (3542)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3423)
1 site
A G T A C T T C A T G A
BsaI  (3004)
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  (2943)
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  (2424)
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  (2316)
1 site
C C C A G C G G G T C G
BseYI  (2312)
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
BglII  (643)
1 site
A G A T C T T C T A G A
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
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

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
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
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
BsgI  (1510)
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  (1641)
1 site
G T A T A C C A T A T G
BstAPI  (1832)
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  (1892)
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  (1892)
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  (2008)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
2870 .. 3730  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2870 .. 3661  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2870 .. 3730  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3662 .. 3730  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2870 .. 3730  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2069 .. 2657  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2069 .. 2657  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
4018 .. 4473  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4018 .. 4473  =  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
CMV promoter
1015 .. 1218  =  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
727 .. 824  =  98 bp
multiple cloning site
MCS
727 .. 824  =  98 bp
multiple cloning site
loxP
2735 .. 2768  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
2735 .. 2768  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
lac UV5 promoter
1576 .. 1607  =  32 bp
   Segment 3:  -10  
   1576 .. 1582  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1576 .. 1607  =  32 bp
   Segment 2:  
   1583 .. 1601  =  19 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1576 .. 1607  =  32 bp
   Segment 1:  -35  
   1602 .. 1607  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1576 .. 1607  =  32 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
691 .. 709  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
691 .. 709  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
832 .. 850  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
832 .. 850  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 fwd
664 .. 680  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
664 .. 680  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
873 .. 889  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
873 .. 889  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
1552 .. 1568  =  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
1552 .. 1568  =  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).
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