pCMV SPORT4

Vector for cloning and transient mammalian cell expression of cDNAs.

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

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HomePlasmidsI.M.A.G.E. Consortium PlasmidspCMV SPORT4
BspDI * - ClaI * (123) NaeI (4258) NgoMIV (4256) DraIII (4155) PflMI * (3843) XmnI (3455) ScaI (3336) BsaI (2917) AhdI (2856) AlwNI (2337) PspFI (2229) BseYI (2225) PciI (1921) HpaI (259) MfeI (268) BsaBI * (360) BclI * (361) NheI (637) BmtI (641) 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) AccI (792) TspMI - XmaI (796) SmaI (798) EcoRI (801) RsrII (808) AgeI (810) Acc65I (813) KpnI (817) PstI - SbfI (822) BfuAI - BspMI (825) SP6 promoter StuI (865) M13 rev SacII (912) AvrII (915) BbsI (944) BsmBI - Esp3I (989) NcoI (1218) SnaBI (1242) NsiI (1529) BstAPI (1745) BspQI - SapI (1805) pCMV SPORT4 4387 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  (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.
PciI  (1921)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (989)
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.
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.
AmpR
2783 .. 3643  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   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
2 segments
   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
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
2648 .. 2681  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
2648 .. 2681  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
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
ORF:  3629 .. 3868  =  240 bp
ORF:  79 amino acids  =  9.0 kDa
ORF:  870 .. 1136  =  267 bp
ORF:  88 amino acids  =  9.5 kDa
ORF:  2913 .. 3179  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1325 .. 1582  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  2783 .. 3643  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Individual Sequences & Maps

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pcDNAII
pCR2.1-TOPO (linearized)
pExpress-1
pAD-GAL4
pcDNA3.1(+)
pCR3.1
pME18S-FL3
pAMP1
pcDNA3.1(-)
pCR3.1 (linearized)
pOTB7
pAMP10
pcDNA3.1 Zeo(+)
pCR4-TOPO
pOTB7-3
pBK-CMV
pCMV SPORT
pCR4-TOPO (linearized)
pOTB7a
pBluescribe (modified)
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pCS105
pPCR-Script Amp SK(+)
pBluescript II SK(+)
pCMV SPORT4
pCS107
pRKW2
pBluescript KS(+)
pCMV SPORT6
pCS108
pSPORT 1
pBluescript SK(+)
pCMV SPORT6.1
pCS2+
pSPORT 2
pBluescript SK(-)
pCMV SPORT6ccdB
pCS22+
pT7T3D-PacI
pBluescript-FL
pCR-Blunt II-TOPO
pCS2G
pUC19
pBluescript II KS(+)
pCR-Blunt II-TOPO (linearized)
pDNR-Dual
pYX-Asc
pBluescriptR
pCR-XL-TOPO
pDNR-LIB
pZErO-2
pcDNA3
pCR-XL-TOPO (linearized)
pDONR201
pZL1
pcDNAI
pCR2.1-TOPO
pDONR223

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