pTrcHis C

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

Sequence Author: Thermo Fisher (Invitrogen)

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PluTI (4284) SfoI (4282) NarI * (4281) KasI (4280) EcoRV (4091) ApaI - BanII (3852) PspOMI (3848) BstEII (3822) BclI * (3655) MluI (3641) PflMI (3223) NsiI (3204) SphI (3202) MauBI (3180) PfoI (3112) PflFI - Tth111I (3013) BsaAI (3007) BstZ17I (2988) AccI (2987) BspQI - SapI (2874) PciI (2757) AlwNI (2348) 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 BsaBI (504) BamHI (513) AvaI - BsoBI - PaeR7I - XhoI (522) BglII (526) PstI (534) Acc65I (538) KpnI (542) EcoRI (550) BstBI (554) HindIII (557) rrnB T2 terminator ScaI (1388) PvuI (1500) FspI (1646) BglI (1751) AhdI (1869) pTrcHis C 4412 bp
PluTI  (4284)
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  (4282)
1 site
G G C G C C C C G C G G
NarI  (4281)
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  (4280)
1 site
G G C G C C C C G C G G
EcoRV  (4091)
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  (3852)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (3852)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PspOMI  (3848)
1 site
G G G C C C C C C G G G
BstEII  (3822)
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  (3655)
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  (3641)
1 site
A C G C G T T G C G C A
PflMI  (3223)
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  (3204)
1 site
A T G C A T T A C G T A
SphI  (3202)
1 site
G C A T G C C G T A C G
MauBI  (3180)
1 site
C G C G C G C G G C G C G C G C
PfoI  (3112)
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  (3013)
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  (3013)
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  (3007)
1 site
Y A C G T R R T G C A Y
BstZ17I  (2988)
1 site
G T A T A C C A T A T G
AccI  (2987)
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  (2874)
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  (2874)
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  (2757)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (2348)
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
BamHI  (513)
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.
AvaI  (522)
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  (522)
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  (522)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (522)
1 site
C T C G A G G A G C T C
BglII  (526)
1 site
A G A T C T T C T A G A
PstI  (534)
1 site
C T G C A G G A C G T C
Acc65I  (538)
1 site
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