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pQCXIN

Self-inactivating bicistronic retroviral vector for expression of a gene together with a neomycin (G418) resistance marker.

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pQCXIN.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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SspI (7055) ScaI (6731) PvuI (6621) AlwNI (5774) SfiI (5064) BmtI (4140) NheI (4136) EcoRV (3994) BsgI (3903) PaeR7I - PspXI - XhoI (3853) AscI (542) PshAI (676) BstEII (1252) Bpu10I (1455) BglII (1576) XbaI (1595) 5' pQC Seq/PCR primer (2141 .. 2164) SbfI (2230) NotI (2243) AgeI (2250) BsiWI (2262) PacI (2273) BamHI (2278) BspEI * (2281) EcoRI (2285) 3' pQC Seq/PCR primer (2329 .. 2352) PspOMI (2421) ApaI (2425) PmlI (2624) AarI (2647) PflMI (2761) BmgBI (2851) BstXI (2894) NgoMIV (3523) NaeI (3525) RsrII (3539) pQCXIN 7400 bp
SspI  (7055)
1 site
A A T A T T T T A T A A
ScaI  (6731)
1 site
A G T A C T T C A T G A
PvuI  (6621)
1 site
C G A T C G G C T A G C
AlwNI  (5774)
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.
SfiI  (5064)
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.
BmtI  (4140)
1 site
G C T A G C C G A T C G
NheI  (4136)
1 site
G C T A G C C G A T C G
EcoRV  (3994)
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.
BsgI  (3903)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PaeR7I  (3853)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3853)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3853)
1 site
C T C G A G G A G C T C
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.
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
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 C C G C C G C C G G C G
AgeI  (2250)
1 site
A C C G G T T G G C C A
BsiWI  (2262)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
PacI  (2273)
1 site
T T A A T T A A A A T T A A T T
BamHI  (2278)
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.
BspEI  (2281)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
EcoRI  (2285)
1 site
G A A T T C C T T A A G
PspOMI  (2421)
1 site
G G G C C C C C C G G G
ApaI  (2425)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PmlI  (2624)
1 site
C A