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pQCXIX

Self-inactivating bicistronic retroviral vector for simultaneous expression of two genes.

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pQCXIX.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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SspI (5964) ScaI (5640) PvuI (5530) FspI (5382) AlwNI (4683) BspQI - SapI (4151) SfiI (3973) AccI (3139) SalI (3138) BmtI (3049) AscI (542) PshAI (676) AfeI (1088) BstEII (1252) Bpu10I (1455) BglII (1576) BspDI * - ClaI * (1588) XbaI (1595) SbfI (2230) NotI (2243) AgeI (2250) BsiWI (2262) PacI (2273) BamHI (2278) BspEI * (2281) EcoRI (2285) PspOMI (2421) ApaI (2425) PmlI (2624) AarI (2647) DraIII (2671) PflMI (2761) BmgBI (2851) BclI * (2885) MluI (2890) PaeR7I - XhoI (2896) EcoRV (2903) PvuII (2946) NheI (3045) pQCXIX 6309 bp
SspI  (5964)
1 site
A A T A T T T T A T A A
ScaI  (5640)
1 site
A G T A C T T C A T G A
PvuI  (5530)
1 site
C G A T C G G C T A G C
FspI  (5382)
1 site
T G C G C A A C G C G T
AlwNI  (4683)
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.
BspQI  (4151)
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  (4151)
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.
SfiI  (3973)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AccI  (3139)
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.
SalI  (3138)
1 site
G T C G A C C A G C T G
BmtI  (3049)
1 site
G C T A G C C G A T C G
AscI  (542)
1 site
G G C G C G C C C C G C G C G G
PshAI  (676)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
AfeI  (1088)
1 site
A G C G C T T C G C G A
BstEII  (1252)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
Bpu10I  (1455)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BglII  (1576)
1 site
A G A T C T T C T A G A
BspDI  (1588)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1588)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
XbaI  (1595)
1 site
T C T A G A A G A T C T
SbfI  (2230)
1 site
C C T G C A G G G G A C G T C C
NotI  (2243)
1 site
G C G G