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pMT-DEST48

Gateway® destination vector for inducible high-level expression of C-terminally V5-6xHis-tagged proteins in Drosophila cells.

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pMT-DEST48.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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EcoO109I (5169) AatII (5115) ZraI (5113) NmeAIII (4341) AhdI (4193) PspFI (3608) BseYI (3604) PciI (3300) BspQI - SapI (3184) SbfI (2933) MfeI (2824) HpaI (2815) PsiI (2795) PmeI (2646) AgeI (2616) MluI (2610) PfoI (46) BstAPI (185) KasI (235) NarI (236) SfoI (237) PluTI (239) BsgI (634) NotI (979) BspEI (1295) EcoRI (1299) PasI - PflMI * (1531) BtgI - NcoI - StyI (1600) BssHII (1793) BstZ17I (1834) BbvCI (2024) AvaI - BsoBI - TspMI - XmaI (2168) BmeT110I (2169) SmaI - SrfI (2170) BstXI (2287) BstBI (2566) BseRI (2583) pMT-DEST48 5179 bp
EcoO109I  (5169)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
AatII  (5115)
1 site
G A C G T C C T G C A G
ZraI  (5113)
1 site
G A C G T C C T G C A G
NmeAIII  (4341)
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  (4193)
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  (3608)
1 site
C C C A G C G G G T C G
BseYI  (3604)
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  (3300)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3184)
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  (3184)
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  (2933)
1 site
C C T G C A G G G G A C G T C C
MfeI  (2824)
1 site
C A A T T G G T T A A C
HpaI  (2815)
1 site
G T T A A C C A A T T G
PsiI  (2795)
1 site
T T A T A A A A T A T T
PmeI  (2646)
1 site
G T T T A A A C C A A A T T T G
AgeI  (2616)
1 site
A C C G G T T G G C C A
MluI  (2610)
1 site
A C G C G T T G C G C A
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.
KasI  (235)
1 site
G G C G C C C C G C G G
NarI  (236)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (237)
1 site
G G C G C C C C G C G G
PluTI  (239)
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.
BsgI  (634)
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).
NotI  (979)
1 site
G C G G C C G C C G C C G G C G
BspEI  (1295)
1 site
T C C G G A A G G C C T
EcoRI  (1299)
1 site
G A A T T C C T T A A G
PasI  (1531)
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  (1531)
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  (1600)
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  (1600)
1 site
C C A T G G G G T A C C
StyI  (1600)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BssHII  (1793)
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  (1834)
1 site
G T A T A C C A T A T G
BbvCI  (2024)
1 site
C C T C A G C G G A G T C G
AvaI  (2168)
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  (2168)
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  (2168)
1 site
C C C G G G G G G C C C
XmaI  (2168)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
BmeT110I  (2169)
1 site
C Y C G R G G R G C Y C
SmaI  (2170)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
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