pRetroX-TetOne

All-in-one retroviral vector for strong inducible expression of genes using the Tet-On® system.

Sequence Author: Clontech (TaKaRa)

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BspQI - SapI (8267) PciI (8150) AlwNI (7741) ScaI (6781) SspI (6457) SacII (5120) AleI (5119) MluI (5092) EcoRV (4734) PflMI (4557) BsiWI (4433) AvrII (77) SnaBI (519) AfeI (717) Eco53kI (733) SacI (735) PshAI (1170) SrfI (1394) AflII (1416) Bsu36I (1428) BstEII (1497) Bpu10I (1700) PpuMI (2077) BamHI (2607) NgoMIV (2613) NaeI (2615) SgrAI (2619) BstZ17I (2627) EcoRI (2633) HindIII (2752) PaeR7I - PspXI - XhoI (3004) AscI (3368) BspEI (3473) NruI (3832) BfuAI - BspMI (4109) pRetroX-TetOne™ 8508 bp
BspQI  (8267)
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  (8267)
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  (8150)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (7741)
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.
ScaI  (6781)
1 site
A G T A C T T C A T G A
SspI  (6457)
1 site
A A T A T T T T A T A A
SacII  (5120)
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.
AleI  (5119)
1 site
C A C N N N N G T G G T G N N N N C A C
MluI  (5092)
1 site
A C G C G T T G C G C A
EcoRV  (4734)
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.
PflMI  (4557)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BsiWI  (4433)
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.
AvrII  (77)
1 site
C C T A G G G G A T C C
SnaBI  (519)
1 site
T A C G T A A T G C A T
AfeI  (717)
1 site
A G C G C T T C G C G A
Eco53kI  (733)
1 site
G A G C T C C T C G A G
SacI  (735)
1 site
G A G C T C C T C G A G
PshAI  (1170)
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.
SrfI  (1394)
1 site
G C C C G G G C C G G G C C C G
AflII  (1416)
1 site
C T T A A G G A A T T C
Bsu36I  (1428)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BstEII  (1497)
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  (1700)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
PpuMI  (2077)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BamHI  (2607)
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.
NgoMIV  (2613)
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.
NaeI  (2615)
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.
SgrAI  (2619)
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.
BstZ17I  (2627)
1 site
G T A T A C C A T A T G
EcoRI  (2633)
1 site
G A A T T C C T T A A G
HindIII  (2752)
1 site
A A G C T T T T C G A A
PaeR7I  (3004)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3004)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3004)
1 site