pMT BioEase-DEST

Gateway® destination vector for inducible high-level expression of biotinylated fusion proteins in Drosophila cells.

Sequence Author: Thermo Fisher (Invitrogen)

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AatII (5311) ZraI (5309) NmeAIII (4537) AhdI (4389) PspFI (3804) BseYI (3800) PciI (3496) BspQI - SapI (3380) SbfI (3129) MfeI (3020) HpaI (3011) PsiI (2991) ApaI (2837) PspOMI (2833) XbaI * (2827) BglII (2824) PacI (2821) PfoI (46) BstAPI (185) BsgI (640) AflII (852) ATG AleI (879) BstEII (881) NotI (1254) BspEI (1570) EcoRI (1574) PasI (1806) MluI (2021) BssHII (2068) NruI (2076) AgeI (2080) BstZ17I (2088) BbvCI (2278) AvaI - BsoBI - TspMI - XmaI (2422) SmaI - SrfI (2424) pMT/BioEase™-DEST 5375 bp
AatII  (5311)
1 site
G A C G T C C T G C A G
ZraI  (5309)
1 site
G A C G T C C T G C A G
NmeAIII  (4537)
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).
AhdI  (4389)
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.
PspFI  (3804)
1 site
C C C A G C G G G T C G
BseYI  (3800)
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.
PciI  (3496)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3380)
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  (3380)
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.
SbfI  (3129)
1 site
C C T G C A G G G G A C G T C C
MfeI  (3020)
1 site
C A A T T G G T T A A C
HpaI  (3011)
1 site
G T T A A C C A A T T G
PsiI  (2991)
1 site
T T A T A A A A T A T T
ApaI  (2837)
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  (2833)
1 site
G G G C C C C C C G G G
XbaI  (2827)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BglII  (2824)
1 site
A G A T C T T C T A G A
PacI  (2821)
1 site
T T A A T T A A A A T T A A T T
PfoI  (46)
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.
BstAPI  (185)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
BsgI  (640)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AflII  (852)
1 site
C T T A A G G A A T T C
AleI  (879)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (881)
1 site
G G T