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pTOL-hCMV-TET3G-Hygro Transactivator

Lentiviral vector with a hygromycin resistance marker for expression of the Tet-On® 3G doxycycline-dependent transactivator.

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pTOL-hCMV-TET3G-Hygro Transactivator.dna
Map and Sequence File:    Download    Open   
Sequence Author:  transOMIC
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SspI (11,153) FspI (10,571) AhdI (10,349) BstZ17I (9080) SexAI * (7496) PmeI (6681) Bsu36I (6288) SgrDI (11,270) KasI (1003) NarI (1004) SfoI (1005) PluTI (1007) BssHII (1077) FseI (1516) EcoNI (1536) BbvCI - Bpu10I (1790) AleI (1943) KflI - PpuMI (2300) BspDI - ClaI (2546) SnaBI (2846) HpaI (3157) BamHI (3176) PaeR7I - XhoI (3939) AarI (4292) BmgBI (4496) MluI (5554) pTOL-hCMV-TET3G-Hygro Transactivator 11,272 bp
SspI  (11,153)
1 site
A A T A T T T T A T A A
FspI  (10,571)
1 site
T G C G C A A C G C G T
AhdI  (10,349)
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.
BstZ17I  (9080)
1 site
G T A T A C C A T A T G
SexAI  (7496)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
PmeI  (6681)
1 site
G T T T A A A C C A A A T T T G
Bsu36I  (6288)
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.
SgrDI  (11,270)
1 site
C G T C G A C G G C A G C T G C
KasI  (1003)
1 site
G G C G C C C C G C G G
NarI  (1004)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (1005)
1 site
G G C G C C C C G C G G
PluTI  (1007)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
BssHII  (1077)
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.
FseI  (1516)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
EcoNI  (1536)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BbvCI  (1790)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1790)
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.
AleI  (1943)
1 site
C A C N N N N G T G G T G N N N N C A C
KflI  (2300)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (2300)
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.
BspDI  (2546)
1 site
A T C G A T T A G C T A
ClaI  (2546)
1 site
A T C G A T T A G C T A
SnaBI  (2846)
1 site
T A C G T A A T G C A T
HpaI  (3157)
1 site
G T T A A C C A A T T G
BamHI  (3176)
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.
PaeR7I  (3939)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (3939)
1 site
C T C G A G G A G C T C
AarI  (4292)
1 site
C A C C