pTrcHis A

Bacterial vector for inducible expression of N-terminally 6xHis-tagged proteins. For other reading frames, use pTrcHis B or pTrcHis C.

Sequence Author: Thermo Fisher (Invitrogen)

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PluTI (4286) SfoI (4284) NarI * (4283) KasI (4282) EcoRV (4093) ApaI (3854) PspOMI (3850) BstEII (3824) BclI * (3657) MluI (3643) PflMI (3225) NsiI (3206) SphI (3204) MauBI (3182) PfoI (3114) PflFI - Tth111I (3015) BsaAI (3009) BstZ17I (2990) AccI (2989) BspQI - SapI (2876) PciI (2759) AlwNI (2350) BfuAI - BspMI (72) trc promoter lac operator gene 10 translational enhancer RBS minicistron BtgI - NcoI - StyI (411) ATG 6xHis NheI (449) BmtI (453) Xpress™ tag BamHI (515) Eco53kI (523) AvaI - BsoBI - PaeR7I - XhoI (524) SacI (525) BglII (528) PstI (536) Acc65I (540) KpnI (544) EcoRI (552) BstBI (556) HindIII (559) rrnB T2 terminator ScaI (1390) PvuI (1502) FspI (1648) BglI (1753) AhdI (1871) pTrcHis A 4414 bp
PluTI  (4286)
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.
SfoI  (4284)
1 site
G G C G C C C C G C G G
NarI  (4283)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (4282)
1 site
G G C G C C C C G C G G
EcoRV  (4093)
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.
ApaI  (3854)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (3850)
1 site
G G G C C C C C C G G G
BstEII  (3824)
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.
BclI  (3657)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
MluI  (3643)
1 site
A C G C G T T G C G C A
PflMI  (3225)
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.
NsiI  (3206)
1 site
A T G C A T T A C G T A
SphI  (3204)
1 site
G C A T G C C G T A C G
MauBI  (3182)
1 site
C G C G C G C G G C G C G C G C
PfoI  (3114)
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.
PflFI  (3015)
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  (3015)
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.
BsaAI  (3009)
1 site
Y A C G T R R T G C A Y
BstZ17I  (2990)
1 site
G T A T A C C A T A T G
AccI  (2989)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
BspQI  (2876)
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  (2876)
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  (2759)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (2350)
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.
BfuAI  (72)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (72)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BtgI  (411)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (411)
1 site
C C A T G G G G T A C C
StyI  (411)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NheI  (449)
1 site
G C T A G C C G A T C G
BmtI  (453)
1 site
G C T A G C C G A T C G
BamHI  (515)
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.
Eco53kI  (523)
1 site
G A G C T C C T C G A G
AvaI  (524)
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  (524)
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  (524)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (524)
1 site
C T C G A G G A G C T C
SacI  (525)
1 site
G A G C T C C T C G A G
BglII  (528)
1 site
A G A T C T T