pMT-DEST48

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

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspMT-DEST48
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) 6xHis 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) 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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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.
SrfI  (2170)
1 site		 G C C C G G G C C G G G C C C G
BstXI  (2287)
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.
BstBI  (2566)
1 site		 T T C G A A A A G C T T
BseRI  (2583)
1 site		 G A G G A G ( N ) 8 N N C T C C T C ( N ) 8
Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
AmpR
4120 .. 4980  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4120 .. 4911  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4120 .. 4980  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4912 .. 4980  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4120 .. 4980  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
1086 .. 1745  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1086 .. 1745  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3361 .. 3949  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3361 .. 3949  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
MT promoter
401 .. 827  =  427 bp
Drosophila metallothionein promoter
MT promoter
401 .. 827  =  427 bp
Drosophila metallothionein promoter
ccdB
2087 .. 2392  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
ccdB
2087 .. 2392  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
SV40 poly(A) signal
2681 .. 2815  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2681 .. 2815  =  135 bp
SV40 polyadenylation signal
attR1
853 .. 977  =  125 bp
recombination site for the Gateway® LR reaction
attR1
853 .. 977  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2433 .. 2557  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2433 .. 2557  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
4981 .. 5085  =  105 bp
AmpR promoter
4981 .. 5085  =  105 bp
V5 tag
2571 .. 2612  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
2571 .. 2612  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
2622 .. 2639  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2622 .. 2639  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lac UV5 promoter
1002 .. 1032  =  31 bp
3 segments
   Segment 1:  -35  
   1002 .. 1007  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1002 .. 1032  =  31 bp
3 segments
   Segment 2:  
   1008 .. 1025  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1002 .. 1032  =  31 bp
3 segments
   Segment 3:  -10  
   1026 .. 1032  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1002 .. 1032  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
ORF:  557 .. 805  =  249 bp
ORF:  82 amino acids  =  9.1 kDa
ORF:  2087 .. 2392  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4250 .. 4516  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  186 .. 440  =  255 bp
ORF:  84 amino acids  =  9.8 kDa
ORF:  1086 .. 1745  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  460 .. 849  =  390 bp
ORF:  129 amino acids  =  14.7 kDa
ORF:  4120 .. 4980  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  399 .. 776  =  378 bp
ORF:  125 amino acids  =  13.2 kDa
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Download pMT-DEST48.dna file

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Individual Sequences & Maps

pAd BLOCK-iT-DEST
pDEST R4-R3 Vector II
pENTR11
pHELLSGATE 4
pAd CMV V5-DEST
pDEST10
pENTR1A
pHELLSGATE 8
pAd PL-DEST
pDEST14
pENTR2B
pIB V5-His-DEST
pBAD-DEST49
pDEST15
pENTR3C
pLenti4 BLOCK-iT-DEST
pcDNA-DEST40
pDEST17
pENTR4
pLenti4 V5-DEST
pcDNA-DEST47
pDEST20
pET-53-DEST
pLenti6.2 C-Lumio V5-DEST
pcDNA-DEST53
pDEST24
pET-54-DEST
pLenti6.2 N-Lumio V5-DEST
pcDNA3.1 nV5-DEST
pDEST26
pET-55-DEST
pLenti6.2 V5-DEST
pcDNA3.2 capTEV-CT V5-DEST
pDEST27
pET-56-DEST
pLenti6.3 TO V5-DEST
pcDNA3.2 capTEV-NT V5-DEST
pDEST8
pET-57-DEST
pLenti6.3 V5-DEST
pcDNA3.2 V5-DEST
pDONR221 P5-P2
pET-58-DEST
pLenti6.4 R4R2 V5-DEST
pcDNA6.2 C-EmGFP-DEST
pDONR P2r-P3
pET-59-DEST
pLenti6 BLOCK-iT-DEST
pcDNA6.2 C-YFP-DEST
pDONR P4-P1r
pET-60-DEST
pLenti6 capTEV-CT-DEST
pcDNA6.2 cGeneBLAzer-DEST
pDONR Zeo
pET-61-DEST
pLenti6 capTEV-NT-DEST1
pcDNA6.2 cLumio-DEST
pDONR201
pET-62-DEST
pLenti6 UbC V5-DEST
pcDNA6.2 EmGFP-Bsd V5-DEST
pDONR207
pET-DEST42
pLenti6 V5-DEST
pcDNA6.2 N-EmGFP-DEST
pDONR221
pET104-DEST
pLenti7.3 V5-DEST
pcDNA6.2 N-YFP-DEST
pDONR221 P1-P4
pET104.1-DEST
pMT-DEST48
pcDNA6.2 nGeneBLAzer-DEST
pDONR221 P1-P5r
pET160-DEST
pMT BioEase-DEST
pcDNA6.2 nLumio-DEST
pDONR221 P3-P2
pET161-DEST
pSCREEN-iT lacZ-DEST
pcDNA6.2 V5-DEST
pDONR221 P4r-P3r
pEXP1-DEST
pT-REx-DEST30
pcDNA6.2 V5-pL-DEST
pDONR221 P5-P4
pEXP2-DEST
pT-REx-DEST31
pCMV SPORT6
pDONR222
pEXP3-DEST
pYES-DEST52
pCMV SPORT6.1
pDONR223
pEXP4-DEST
Reading Frame Cassette A
pCOLA-2-DEST
pEF-DEST51
pHELLSGATE
Reading Frame Cassette B
pCOLA-3-DEST
pEF5 FRT V5-DEST
pHELLSGATE 12
Reading Frame Cassette C.1
pDEST R4-R3
pENTR GeneBLAzer

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