pTrcHis B

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

Sequence Author: Thermo Fisher (Invitrogen)

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PluTI (4276) SfoI (4274) NarI * (4273) KasI (4272) HpaI (4139) EcoRV (4083) ApaI (3844) PspOMI (3840) BstEII (3814) BclI * (3647) MluI (3633) PflMI (3215) NsiI (3196) SphI (3194) MauBI (3172) PfoI (3104) PflFI - Tth111I (3005) BsaAI (2999) BstZ17I (2980) AccI (2979) BspQI - SapI (2866) PciI (2749) AlwNI (2340) BfuAI - BspMI (72) trc promoter lac operator gene 10 translational enhancer RBS minicistron BtgI - NcoI - StyI (411) ATG NheI (449) BmtI (453) T7 tag (gene 10 leader) Xpress™ tag BsaBI * (504) BamHI (505) Eco53kI (513) AvaI - BsoBI - PaeR7I - XhoI (514) SacI (515) BglII (518) PstI (526) Acc65I (530) KpnI (534) EcoRI (542) BstBI (546) HindIII (549) ScaI (1380) PvuI (1492) FspI (1638) BglI (1743) AhdI (1861) pTrcHis B 4404 bp
PluTI  (4276)
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  (4274)
1 site
G G C G C C C C G C G G
NarI  (4273)
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  (4272)
1 site
G G C G C C C C G C G G
HpaI  (4139)
1 site
G T T A A C C A A T T G
EcoRV  (4083)
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  (3844)
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  (3840)
1 site
G G G C C C C C C G G G
BstEII  (3814)
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  (3647)
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  (3633)
1 site
A C G C G T T G C G C A
PflMI  (3215)
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  (3196)
1 site
A T G C A T T A C G T A
SphI  (3194)
1 site
G C A T G C C G T A C G
MauBI  (3172)
1 site
C G C G C G C G G C G C G C G C
PfoI  (3104)
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  (3005)
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  (3005)
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  (2999)
1 site
Y A C G T R R T G C A Y
BstZ17I  (2980)
1 site
G T A T A C C A T A T G
AccI  (2979)
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  (2866)
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  (2866)
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  (2749)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (2340)
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
BsaBI  (504)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BamHI  (505)
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  (513)
1 site
G A G C T C C T C G A G
AvaI  (514)
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  (514)
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  (514)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (514)
1 site
C T C G A G G A G C T C
SacI  (515)
1 site
G A G C T C C T C G A G
BglII  (518)
1 site
A G A T C T T C T A G A
PstI  (526)
1 site
C T G C A G G A C G T C
Acc65I  (530)
1 site
G G T A C C C C A T G G
KpnI  (534)
1 site
G G T A C C C C A T G G
EcoRI  (542)
1 site
G A A T T C C T T A A G
BstBI