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 See pCMV SPORT6.1 for a newer version of this vector.

Sequence Author: Thermo Fisher (Invitrogen)

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HindIII (67) 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) 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 - Esp3I (323) NcoI (552) SnaBI (576) NsiI (863) BstAPI (1079) BspQI - SapI (1139) PciI (1255) AlwNI (1671) AhdI (2190) pCMV SPORT6 4396 bp
HindIII  (67)
1 site
A A G C T T T T C G A A
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.
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

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (323)
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  (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
2 segments
   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
2 segments
   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
ORF:  2963 .. 3202  =  240 bp
ORF:  79 amino acids  =  9.0 kDa
ORF:  204 .. 470  =  267 bp
ORF:  88 amino acids  =  9.5 kDa
ORF:  2247 .. 2513  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  659 .. 916  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  2117 .. 2977  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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