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pCMV SPORT6ccdB

Gateway® library vector for low-background cloning and transient mammalian cell expression of cDNAs.

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pCMV SPORT6ccdB.dna
Map and Sequence File:    Download    Open   
Sequence Author:  I.M.A.G.E. Consortium
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NaeI (4678) NgoMIV (4676) DraIII (4575) PflMI * (4263) XmnI (3875) ScaI (3756) AhdI (3276) AlwNI (2757) PciI (2341) BspDI * - ClaI * (123) HpaI (259) MfeI (268) BclI * (361) NheI (637) BmtI (641) MluI (736) HindIII (742) PspOMI (748) ApaI (752) AbsI - PaeR7I - PspXI - XhoI (754) BamHI (760) EagI - NotI (773) BssHII (821) BstZ17I (862) SrfI (982) BmgBI (1016) BstXI (1099) SalI (1205) EcoRV (1213) EcoRI (1221) RsrII (1228) AgeI (1230) Acc65I (1233) KpnI (1237) StuI (1285) SacII (1332) AvrII (1335) BbsI (1364) BsmBI (1409) Eco53kI (1436) SacI (1438) NcoI (1638) SnaBI (1662) NsiI (1949) BstAPI (2165) BspQI - SapI (2225) pCMV SPORT6ccdB 4807 bp
NaeI  (4678)
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  (4676)
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  (4575)
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  (4263)
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  (3875)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3756)
1 site
A G T A C T T C A T G A
AhdI  (3276)
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  (2757)
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.
PciI  (2341)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
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.
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
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
MluI  (736)
1 site
A C G C G T T G C G C A
HindIII  (742)
1 site
A A G C T T T T C G A A
PspOMI  (748)
1 site
G G G C C C C C C G G G
ApaI  (752)
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  (754)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (754)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (754)
1 site
V C T C G A G B B G A G C T C V
XhoI  (754)
1 site
C T C G A G G A G C T C
BamHI  (760)
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.
EagI  (773)
1 site
C G G C C G G C C G G C
NotI  (773)
1 site
G C G G C C G C C G C C G G C G
BssHII  (821)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BstZ17I  (862)
1 site
G T A T A C C A T A T G
SrfI  (982)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (1016)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
BstXI  (1099)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
SalI  (1205)
1 site
G T C G A C C A G C T G
EcoRV  (1213)
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.
EcoRI  (1221)
1 site
G A A T T C C T T A A G
RsrII  (1228)
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  (1230)
1 site
A C C G G T T G G C C A
Acc65I  (1233)
1 site
G G T A C C C C A T G G
KpnI  (1237)
1 site
G G T A C C C C A T G G
StuI  (1285)
1 site
A G G C C T T C C G G A
SacII  (1332)
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  (1335)
1 site<