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pQC-tTS-IN

Retroviral vector for expressing tTS, which represses expression from Tet-responsive promoters in the absence of doxycycline.

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pQC-tTS-IN.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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ScaI (7557) PvuI (7447) AlwNI (6600) StuI (5936) SfiI (5890) EcoRV (4820) PaeR7I - PspXI - XhoI (4679) RsrII (4365) NaeI (4351) NgoMIV (4349) AscI (542) PshAI (676) BstEII (1252) Bpu10I (1455) BglII (1576) SbfI (2230) NotI (2243) AgeI - SgrAI (2250) BamHI (3098) EcoRI (3111) PspOMI (3247) ApaI (3251) AarI (3473) PflMI (3587) BmgBI (3677) pQC-tTS-IN 8226 bp
ScaI  (7557)
1 site
A G T A C T T C A T G A
PvuI  (7447)
1 site
C G A T C G G C T A G C
AlwNI  (6600)
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.
StuI  (5936)
1 site
A G G C C T T C C G G A
SfiI  (5890)
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.
EcoRV  (4820)
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.
PaeR7I  (4679)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4679)
1 site
V C T C G A G B B G A G C T C V
XhoI  (4679)
1 site
C T C G A G G A G C T C
RsrII  (4365)
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.
NaeI  (4351)
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  (4349)
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.
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
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
SgrAI  (2250)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
BamHI  (3098)
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.
EcoRI  (3111)
1 site
G A A T T C C T T A A G
PspOMI  (3247)
1 site
G G G C C C C C C G G G
ApaI  (3251)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AarI  (3473)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4