pDEST20

Gateway® destination vector for expressing N-terminally GST-tagged proteins in insect cells using the Bac-to-Bac® baculovirus system.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspDEST20
BbsI (6946) PflFI - Tth111I (6417) BseRI (6150) EcoRV (6014) SacII (5957) PspFI (5153) BseYI (5149) FspI (4342) PvuI (4196) BsaHI (4025) BanII (3350) SnaBI (4) EcoNI (171) EcoO109I (193) BsgI (447) BstBI (558) SwaI (588) NotI (968) PvuII (1188) EcoRI (1288) PasI - PflMI * (1520) NcoI (1589) XbaI (1735) BssHII (1782) BbvCI (2013) TspMI - XmaI (2157) SmaI - SrfI (2159) BmgBI (2193) BfuAI - BspMI (2403) PstI (2414) SalI (2416) MfeI (2666) HpaI (2679) AvrII (2830) NgoMIV (3316) NaeI (3318) pDEST™20 7066 bp
BbsI  (6946)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
PflFI  (6417)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (6417)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BseRI  (6150)
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.
EcoRV  (6014)
1 site		 G A T A T C C T A T A G
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
SacII  (5957)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
PspFI  (5153)
1 site		 C C C A G C G G G T C G
BseYI  (5149)
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.
FspI  (4342)
1 site		 T G C G C A A C G C G T
PvuI  (4196)
1 site		 C G A T C G G C T A G C
BsaHI  (4025)
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.
BanII  (3350)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
SnaBI  (4)
1 site		 T A C G T A A T G C A T
EcoNI  (171)
1 site		 C C T N N N N N A G G G G A N N N N N T C C
The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
EcoO109I  (193)
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.
BsgI  (447)
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).
BstBI  (558)
1 site		 T T C G A A A A G C T T
SwaI  (588)
1 site		 A T T T A A A T T A A A T T T A
SwaI is typically used at 25°C, but is 50% active at 37°C.
NotI  (968)
1 site		 G C G G C C G C C G C C G G C G
PvuII  (1188)
1 site		 C A G C T G G T C G A C
EcoRI  (1288)
1 site		 G A A T T C C T T A A G
PasI  (1520)
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  (1520)
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.
NcoI  (1589)
1 site		 C C A T G G G G T A C C
XbaI  (1735)
1 site		 T C T A G A A G A T C T
BssHII  (1782)
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.
BbvCI  (2013)
1 site		 C C T C A G C G G A G T C G
TspMI  (2157)
1 site		 C C C G G G G G G C C C
XmaI  (2157)
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  (2159)
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  (2159)
1 site		 G C C C G G G C C G G G C C C G
BmgBI  (2193)
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.
BfuAI  (2403)
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  (2403)
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 BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PstI  (2414)
1 site		 C T G C A G G A C G T C
SalI  (2416)
1 site		 G T C G A C C A G C T G
MfeI  (2666)
1 site		 C A A T T G G T T A A C
HpaI  (2679)
1 site		 G T T A A C C A A T T G
AvrII  (2830)
1 site		 C C T A G G G G A T C C
NgoMIV  (3316)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (3318)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
AmpR
3778 .. 4638  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3778 .. 3846  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3778 .. 4638  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3847 .. 4638  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3778 .. 4638  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
1075 .. 1734  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1075 .. 1734  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
GST
161 .. 814  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
161 .. 814  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
ori
4809 .. 5397  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4809 .. 5397  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
GmR
5991 .. 6524  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
5991 .. 6524  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
f1 ori
3191 .. 3646  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3191 .. 3646  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
ccdB
2076 .. 2381  =  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
2076 .. 2381  =  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.
Tn7R
5700 .. 5924  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7R
5700 .. 5924  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7L
2843 .. 3008  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
Tn7L
2843 .. 3008  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
SV40 poly(A) signal
2680 .. 2814  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2680 .. 2814  =  135 bp
SV40 polyadenylation signal
attR1
842 .. 966  =  125 bp
recombination site for the Gateway® LR reaction
attR1
842 .. 966  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2422 .. 2546  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2422 .. 2546  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
3673 .. 3777  =  105 bp
AmpR promoter
3673 .. 3777  =  105 bp
polyhedrin promoter
27 .. 118  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
27 .. 118  =  92 bp
promoter for the baculovirus polyhedrin gene
lac promoter
991 .. 1021  =  31 bp
3 segments
   Segment 1:  -35  
   991 .. 996  =  6 bp
promoter for the E. coli lac operon
lac promoter
991 .. 1021  =  31 bp
3 segments
   Segment 2:  
   997 .. 1014  =  18 bp
promoter for the E. coli lac operon
lac promoter
991 .. 1021  =  31 bp
3 segments
   Segment 3:  -10  
   1015 .. 1021  =  7 bp
promoter for the E. coli lac operon
lac promoter
991 .. 1021  =  31 bp
3 segments
promoter for the E. coli lac operon
Pc promoter
6713 .. 6741  =  29 bp
3 segments
   Segment 3:  -10  
   6713 .. 6718  =  6 bp
class 1 integron promoter
Pc promoter
6713 .. 6741  =  29 bp
3 segments
   Segment 2:  
   6719 .. 6735  =  17 bp
class 1 integron promoter
Pc promoter
6713 .. 6741  =  29 bp
3 segments
   Segment 1:  -35  
   6736 .. 6741  =  6 bp
class 1 integron promoter
Pc promoter
6713 .. 6741  =  29 bp
3 segments
class 1 integron promoter
ORF:  1075 .. 1734  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3778 .. 4638  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  161 .. 877  =  717 bp
ORF:  238 amino acids  =  27.9 kDa
ORF:  5834 .. 6136  =  303 bp
ORF:  100 amino acids  =  11.8 kDa
ORF:  6626 .. 39  =  480 bp
ORF:  159 amino acids  =  18.5 kDa
ORF:  2076 .. 2381  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  6129 .. 6461  =  333 bp
ORF:  110 amino acids  =  11.9 kDa
ORF:  6735 .. 6977  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  6716 .. 7024  =  309 bp
ORF:  102 amino acids  =  11.4 kDa
ORF:  4242 .. 4508  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5991 .. 6524  =  534 bp
ORF:  177 amino acids  =  19.4 kDa
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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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I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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