TAGZyme pQE-2

Vector with a lacIq repressor gene for bacterial expression of 6xHis-tagged proteins that can be processed by TAGZyme.

Sequence Author: Qiagen

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T5 promoter AvaI - BsoBI - PaeR7I - XhoI (1) AatII (4688) ZraI (4686) XmnI (4365) PvuI (4136) BglI (3886) BsaI (3827) AhdI (3766) AlwNI (3289) PciI (2873) BspQI - SapI (2757) BstZ17I (2646) PflFI - Tth111I (2620) PfoI (2517) FspAI (2453) PpuMI * (2436) PsiI (49) MfeI (59) lac operator EcoRI (88) RBS BseRI (115) ATG 6xHis NdeI (140) NsiI (146) SphI (148) Eco53kI (154) SacI (156) EagI - NotI (161) SacII (167) Acc65I (169) KpnI (173) PmlI (177) SalI (180) PstI (190) BfuAI - BspMI (193) HindIII (194) BlpI (206) NheI (314) BmtI (318) Bpu10I (339) BspEI (562) PasI (798) NcoI - StyI (867) KasI (1344) NarI * (1345) SfoI (1346) PluTI (1348) HpaI (1481) EcoRV (1537) BssHII (1572) PspOMI (1776) ApaI (1780) BstEII (1801) BclI * (1969) MluI (1983) TAGZyme pQE-2 4758 bp
AvaI  (1)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (1)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (1)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1)
1 site
C T C G A G G A G C T C
AatII  (4688)
1 site
G A C G T C C T G C A G
ZraI  (4686)
1 site
G A C G T C C T G C A G
XmnI  (4365)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (4136)
1 site
C G A T C G G C T A G C
BglI  (3886)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
BsaI  (3827)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (3766)
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.
AlwNI  (3289)
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.
PciI  (2873)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2757)
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  (2757)
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.
BstZ17I  (2646)
1 site
G T A T A C C A T A T G
PflFI  (2620)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2620)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PfoI  (2517)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
FspAI  (2453)
1 site
R T G C G C A Y Y A C G C G T R
PpuMI  (2436)
1 site
R G G W C C Y Y C C W G G R
* Blocked by Dcm methylation.
Sticky ends from different PpuMI sites may not be compatible.
PsiI  (49)
1 site
T T A T A A A A T A T T
MfeI  (59)
1 site
C A A T T G G T T A A C
EcoRI  (88)
1 site
G A A T T C C T T A A G
BseRI  (115)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
NdeI  (140)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
NsiI  (146)
1 site
A T G C A T T A C G T A
SphI  (148)
1 site
G C A T G C C G T A C G
Eco53kI  (154)
1 site
G A G C T C C T C G A G
SacI  (156)
1 site
G A G C T C C T C G A G
EagI  (161)
1 site
C G G C C G G C C G G C
NotI  (161)
1 site
G C G G C C G C C G C C G G C G
SacII  (167)
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.
Acc65I  (169)
1 site
G G T A C C C C A T G G
KpnI  (173)
1 site
G G T A C C C C A T