pEXP1-DEST

Gateway® destination vector for cell-free expression and N-terminal tagging. See also pEXP3-DEST.

Sequence Author: Thermo Fisher (Invitrogen)

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BspQI - SapI (4560) NspI (4447) AflIII - PciI (4443) PspFI (4143) BseYI (4139) AhdI (3555) AfeI (3524) BmrI (3515) NmeAIII (3408) BsaHI (3015) XmnI (2955) PsiI (2485) DraIII (2360) BsaAI (2357) BglII (0) RBS NdeI (98) ATG 6xHis NheI (136) BmtI (140) Xpress™ tag EagI - NotI (328) lac UV5 promoter PvuII (548) EcoRI (648) BsmBI - Esp3I (872) PasI - PflMI * (880) BtgI - NcoI (949) BssHII (1142) BstZ17I (1183) BbvCI (1373) AvaI - BsoBI - TspMI - XmaI (1517) SmaI - SrfI (1519) BmgBI (1553) BstXI (1636) BfuAI - BspMI (1763) PstI (1774) SalI (1776) HindIII (1914) BlpI (1972) EcoO109I (1999) NgoMIV (2252) NaeI (2254) BanII (2286) pEXP1-DEST 4622 bp
BspQI  (4560)
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  (4560)
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.
NspI  (4447)
1 site
R C A T G Y Y G T A C R
AflIII  (4443)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4443)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (4143)
1 site
C C C A G C G G G T C G
BseYI  (4139)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AhdI  (3555)
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.
AfeI  (3524)
1 site
A G C G C T T C G C G A
BmrI  (3515)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
NmeAIII  (3408)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsaHI  (3015)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
XmnI  (2955)
1 site
G A A N N N N T T C C T T N N N N A A G
PsiI  (2485)
1 site
T T A T A A A A T A T T
DraIII  (2360)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BsaAI  (2357)
1 site
Y A C G T R R T G C A Y
BglII  (0)
1 site
A G A T C T T C T A G A
NdeI  (98)
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.
NheI  (136)
1 site
G C T A G C C G A T C G
BmtI  (140)
1 site
G C T A G C C G A T C G
EagI  (328)
1 site
C G G C C G G C C G G C
NotI  (328)
1 site
G C G G C C G C C G C C G G C G
PvuII  (548)
1 site
C A G C T G G T C G A C
EcoRI  (648)
1 site
G A A T T C C T T A A G
BsmBI  (872)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (872)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
PasI  (880)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (880)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BtgI  (949)
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  (949)
1 site
C C A T G G G G T A C C
BssHII  (1142)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BstZ17I  (1183)
1 site
G T A T A C C A T A T G
BbvCI  (1373)
1 site
C C T C A G C G G A G T C G
AvaI  (1517)
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  (1517)
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.
TspMI  (1517)
1 site
C C C G G G G G G C C C
XmaI  (1517)
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  (1519)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (1519)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (1553)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
BstXI  (1636)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BfuAI  (1763)
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  (1763)
1 site
A C C T G C ( N