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Plasmid Files

pCMV SPORT6

Gateway® library vector for cloning and transient mammalian cell expression of cDNAs. See pCMV SPORT6.1 for a newer version of this vector.

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pCMV SPORT6 Sequence and MappCMV SPORT6.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 MluI (61) T7 promoter M13 fwd BmtI (4362) NheI (4358) BclI * (4082) BsaBI * (4081) MfeI (3989) HpaI (3980) BspDI * - ClaI * (3844) NaeI (3592) NgoMIV (3590) DraIII (3489) PflMI * (3177) XmnI (2789) ScaI (2670) BsaI (2251) HindIII (67) PspOMI (73) ApaI (77) AbsI - PaeR7I - PspXI - XhoI (79) XbaI (91) EagI - NotI (98) SpeI (107) SalI - SgrDI (119) AccI (120) EcoRV (127) TspMI - XmaI (130) SmaI (132) EcoRI (135) RsrII (142) AgeI (144) Acc65I (147) KpnI (151) SP6 promoter StuI (199) M13 rev SacII (246) AvrII (249) BbsI (278) BsmBI (323) NcoI (552) SnaBI (576) CMV enhancer NsiI (863) BstAPI (1079) BspQI - SapI (1139) PciI (1255) AlwNI (1671) AhdI (2190) pCMV SPORT6 4396 bp
MluI  (61)
1 site
A C G C G T T G C G C A
BmtI  (4362)
1 site
G C T A G C C G A T C G
NheI  (4358)
1 site
G C T A G C C G A T C G
BclI  (4082)
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.
BsaBI  (4081)
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.
MfeI  (3989)
1 site
C A A T T G G T T A A C
HpaI  (3980)
1 site
G T T A A C C A A T T G
BspDI  (3844)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3844)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
NaeI  (3592)
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  (3590)
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  (3489)
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  (3177)
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  (2789)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2670)
1 site
A G T A C T T C A T G A
BsaI  (2251)
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.
HindIII  (67)
1 site
A A G C T T T T C G A A
PspOMI  (73)
1 site
G G G C C C C C C G G G
ApaI  (77)
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  (79)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (79)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (79)
1 site
V C T C G A G B B G A G C T C V
XhoI  (79)
1 site
C T C G A G G A G C T C
XbaI  (91)
1 site
T C T A G A A G A T C T
EagI  (98)
1 site
C G G C C G G C C G G C
NotI  (98)
1 site
G C G G C C G C C G C C G G C G
SpeI  (107)
1 site
A C T A G T T G A T C A
SalI  (119)
1 site
G T C G A C C A G C T G
SgrDI  (119)
1 site
C G T C G A C G G C A G C T G C
AccI  (120)
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.
EcoRV  (127)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
TspMI  (130)
1 site
C C C G G G G G G C C C
XmaI  (130)
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  (132)
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  (135)
1 site
G A A T T C C T T A A G
RsrII  (142)
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  (144)
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  (147)
1 site
G G T A C C C C A T G G
KpnI  (151)
1 site
G G T A C C C C A T G G
StuI  (199)
1 site
A G G C C T T C C G G A
SacII  (246)
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  (249)
1 site
C C T A G G G G A T C C
BbsI  (278)
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  (323)
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  (552)
1 site
C C A T G G G G T A C C
SnaBI  (576)
1 site
T A C G T A A T G C A T
NsiI  (863)
1 site
A T G C A T T A C G T A
BstAPI  (1079)
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  (1139)
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  (1139)
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  (1255)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (1671)
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.
AhdI  (2190)
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.
AmpR
2117 .. 2977  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2117 .. 2908  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2117 .. 2977  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2909 .. 2977  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2117 .. 2977  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1316 .. 1904  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1316 .. 1904  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3265 .. 3720  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3265 .. 3720  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
CMV enhancer
553 .. 856  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
553 .. 856  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
349 .. 552  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
349 .. 552  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
3846 .. 3980  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3846 .. 3980  =  135 bp
SV40 polyadenylation signal
loxP
1982 .. 2015  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
1982 .. 2015  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
attB2
36 .. 60  =  25 bp
recombination site for the Gateway® BP reaction
attB2
36 .. 60  =  25 bp
recombination site for the Gateway® BP reaction
attB1
153 .. 177  =  25 bp
recombination site for the Gateway® BP reaction
attB1
153 .. 177  =  25 bp
recombination site for the Gateway® BP reaction
T7 promoter
16 .. 34  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
16 .. 34  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
178 .. 196  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
178 .. 196  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 rev
207 .. 223  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
207 .. 223  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
4385 .. 5  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
4385 .. 5  =  17 bp
common sequencing primer, one of multiple similar
variants
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