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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HomePlasmidsGateway® Cloning VectorspMT BioEase-DEST
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 N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
NotI  (1254)
1 site		 G C G G C C G C C G C C G G C G
BspEI  (1570)
1 site		 T C C G G A A G G C C T
EcoRI  (1574)
1 site		 G A A T T C C T T A A G
PasI  (1806)
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.
MluI  (2021)
1 site		 A C G C G T T G C G C A
BssHII  (2068)
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.
NruI  (2076)
1 site		 T C G C G A A G C G C T
AgeI  (2080)
1 site		 A C C G G T T G G C C A
BstZ17I  (2088)
1 site		 G T A T A C C A T A T G
BbvCI  (2278)
1 site		 C C T C A G C G G A G T C G
AvaI  (2422)
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  (2422)
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  (2422)
1 site		 C C C G G G G G G C C C
XmaI  (2422)
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  (2424)
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  (2424)
1 site		 G C C C G G G C C G G G C C C G
AmpR
4316 .. 5176  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4316 .. 5107  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4316 .. 5176  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5108 .. 5176  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4316 .. 5176  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
1361 .. 2020  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1361 .. 2020  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3557 .. 4145  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3557 .. 4145  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
MT promoter
407 .. 833  =  427 bp
Drosophila metallothionein promoter
MT promoter
407 .. 833  =  427 bp
Drosophila metallothionein promoter
ccdB
2341 .. 2646  =  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
2341 .. 2646  =  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.
ATG
864 .. 866  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
864 .. 866  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
BioEase™ tag
867 .. 1082  =  216 bp
72 amino acids  =  7.1 kDa
3 segments
   Segment 1:  
   867 .. 977  =  111 bp
   37 amino acids  =  3.6 kDa
Product: biotinylated sequence derived from the C-terminus of the α-subunit of Klebsiella pneumoniae oxaloacetate decarboxylase
contains a unique biotinylation site
BioEase™ tag
867 .. 1082  =  216 bp
72 amino acids  =  7.1 kDa
3 segments
   Segment 2:  site  
   978 .. 980  =  3 bp
   1 amino acid  =  146.2 Da
Product: biotinylated sequence derived from the C-terminus of the α-subunit of Klebsiella pneumoniae oxaloacetate decarboxylase
contains a unique biotinylation site
BioEase™ tag
867 .. 1082  =  216 bp
72 amino acids  =  7.1 kDa
3 segments
   Segment 3:  
   981 .. 1082  =  102 bp
   34 amino acids  =  3.4 kDa
Product: biotinylated sequence derived from the C-terminus of the α-subunit of Klebsiella pneumoniae oxaloacetate decarboxylase
contains a unique biotinylation site
BioEase™ tag
867 .. 1082  =  216 bp
72 amino acids  =  7.1 kDa
3 segments
Product: biotinylated sequence derived from the C-terminus of the α-subunit of Klebsiella pneumoniae oxaloacetate decarboxylase
contains a unique biotinylation site
Xpress™ tag
1095 .. 1118  =  24 bp
8 amino acids  =  998.0 Da
2 segments
   Segment 1:  
   1095 .. 1103  =  9 bp
   3 amino acids  =  409.4 Da
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
Xpress™ tag
1095 .. 1118  =  24 bp
8 amino acids  =  998.0 Da
2 segments
   Segment 2:  enterokinase site  
   1104 .. 1118  =  15 bp
   5 amino acids  =  606.5 Da
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
Xpress™ tag
1095 .. 1118  =  24 bp
8 amino acids  =  998.0 Da
2 segments
Product: Xpress™ epitope tag, including an enterokinase recognition and cleavage site
SV40 poly(A) signal
2877 .. 3011  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2877 .. 3011  =  135 bp
SV40 polyadenylation signal
attR1
1128 .. 1252  =  125 bp
recombination site for the Gateway® LR reaction
attR1
1128 .. 1252  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2687 .. 2811  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2687 .. 2811  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
5177 .. 5281  =  105 bp
AmpR promoter
5177 .. 5281  =  105 bp
lac UV5 promoter
1277 .. 1307  =  31 bp
3 segments
   Segment 1:  -35  
   1277 .. 1282  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1277 .. 1307  =  31 bp
3 segments
   Segment 2:  
   1283 .. 1300  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1277 .. 1307  =  31 bp
3 segments
   Segment 3:  -10  
   1301 .. 1307  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1277 .. 1307  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
ORF:  2341 .. 2646  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  563 .. 811  =  249 bp
ORF:  82 amino acids  =  9.2 kDa
ORF:  1361 .. 2020  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  186 .. 446  =  261 bp
ORF:  86 amino acids  =  9.9 kDa
ORF:  864 .. 1163  =  300 bp
ORF:  99 amino acids  =  10.2 kDa
ORF:  4446 .. 4712  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  405 .. 782  =  378 bp
ORF:  125 amino acids  =  13.3 kDa
ORF:  840 .. 1124  =  285 bp
ORF:  94 amino acids  =  10.3 kDa
ORF:  4316 .. 5176  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  466 .. 816  =  351 bp
ORF:  116 amino acids  =  13.3 kDa
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Download pMT BioEase-DEST.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

Plasmid Sets

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TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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