pTriEx-1.1

Insect, bacterial, and mammalian vector for expressing proteins with a C-terminal HSV-8xHis cassette.

Sequence Author: MilliporeSigma (Novagen)

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AanI (3610) AanI (3053) AanI (320) AanI (409) CMV enhancer NcoI (2331) EcoRV (2340) TspMI - XmaI (2343) SmaI (2345) Eco53kI (2350) SacI (2352) BamHI (2356) EcoRI (2362) BglII (2370) BfuAI - BspMI (2377) AscI (2378) PstI - SbfI (2388) Acc65I (2396) AgeI (2399) KpnI (2400) BstBI (2405) HindIII (2408) NotI (2415) PvuII (2425) BstZ17I (2430) PmlI (2435) PaeR7I - XhoI (2477) DraIII (2509) pTriEx™-1.1 5301 bp
AanI  (3610)
4 sites
T T A T A A A A T A T T
AanI  (3053)
4 sites
T T A T A A A A T A T T
AanI  (320)
4 sites
T T A T A A A A T A T T
AanI  (409)
4 sites
T T A T A A A A T A T T
NcoI  (2331)
1 site
C C A T G G G G T A C C
EcoRV  (2340)
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.
TspMI  (2343)
1 site
C C C G G G G G G C C C
XmaI  (2343)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2345)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Eco53kI  (2350)
1 site
G A G C T C C T C G A G
SacI  (2352)
1 site
G A G C T C C T C G A G
BamHI  (2356)
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.
EcoRI  (2362)
1 site
G A A T T C C T T A A G
BglII  (2370)
1 site
A G A T C T T C T A G A
BfuAI  (2377)
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  (2377)
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.
AscI  (2378)
1 site
G G C G C G C C C C G C G C G G
PstI  (2388)
1 site