pcDNA-DEST40

Gateway® destination vector for expressing C-terminally V5- and 6xHis-tagged proteins in mammalian cells.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspcDNA-DEST40
SgrDI (7141) PvuI (6590) AhdI (6220) PciI (5327) PfoI (4751) RsrII (4490) PflFI - Tth111I (4092) PluTI (3977) SfoI (3975) NarI (3974) KasI (3973) AvrII (3764) StuI (3763) MfeI (161) SpeI (249) CMV enhancer NdeI (484) SnaBI (590) Eco53kI (816) SacI (818) NotI (1037) lac UV5 promoter BspEI (1353) EcoRI (1357) BsmBI - Esp3I (1581) PasI - PflMI * (1589) BbvCI (2082) SrfI (2228) BmgBI (2262) BstXI (2345) XbaI * (2619) EcoO109I - PspOMI (2625) ApaI (2629) SacII (2632) BstBI (2636) AgeI (2686) 6xHis PmeI (2716) BbsI (2927) DraIII (3241) CsiI - SexAI * (3531) pcDNA™-DEST40 7143 bp
SgrDI  (7141)
1 site		 C G T C G A C G G C A G C T G C
PvuI  (6590)
1 site		 C G A T C G G C T A G C
AhdI  (6220)
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.
PciI  (5327)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
PfoI  (4751)
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.
RsrII  (4490)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
PflFI  (4092)
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  (4092)
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.
PluTI  (3977)
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.
SfoI  (3975)
1 site		 G G C G C C C C G C G G
NarI  (3974)
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.
KasI  (3973)
1 site		 G G C G C C C C G C G G
AvrII  (3764)
1 site		 C C T A G G G G A T C C
StuI  (3763)
1 site		 A G G C C T T C C G G A
MfeI  (161)
1 site		 C A A T T G G T T A A C
SpeI  (249)
1 site		 A C T A G T T G A T C A
NdeI  (484)
1 site		 C A T A T G G T A T A C
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
SnaBI  (590)
1 site		 T A C G T A A T G C A T
Eco53kI  (816)
1 site		 G A G C T C C T C G A G
SacI  (818)
1 site		 G A G C T C C T C G A G
NotI  (1037)
1 site		 G C G G C C G C C G C C G G C G
BspEI  (1353)
1 site		 T C C G G A A G G C C T
EcoRI  (1357)
1 site		 G A A T T C C T T A A G
BsmBI  (1581)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (1581)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
PasI  (1589)
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  (1589)
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.
BbvCI  (2082)
1 site		 C C T C A G C G G A G T C G
SrfI  (2228)
1 site		 G C C C G G G C C G G G C C C G
BmgBI  (2262)
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.
BstXI  (2345)
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.
XbaI  (2619)
1 site		 T C T A G A A G A T C T
* Blocked by Dam methylation.
EcoO109I  (2625)
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.
PspOMI  (2625)
1 site		 G G G C C C C C C G G G
ApaI  (2629)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
SacII  (2632)
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.
BstBI  (2636)
1 site		 T T C G A A A A G C T T
AgeI  (2686)
1 site		 A C C G G T T G G C C A
PmeI  (2716)
1 site		 G T T T A A A C C A A A T T T G
BbsI  (2927)
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.
DraIII  (3241)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
CsiI  (3531)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (3531)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
AmpR
6147 .. 7007  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6147 .. 6938  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6147 .. 7007  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6939 .. 7007  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6147 .. 7007  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
3846 .. 4640  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
3846 .. 4640  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
CmR
1144 .. 1803  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1144 .. 1803  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
5388 .. 5976  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5388 .. 5976  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3008 .. 3436  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3008 .. 3436  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
3450 .. 3779  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3450 .. 3779  =  330 bp
SV40 enhancer and early promoter
ccdB
2145 .. 2450  =  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
2145 .. 2450  =  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.
bGH poly(A) signal
2738 .. 2962  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
2738 .. 2962  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
attR1
911 .. 1035  =  125 bp
recombination site for the Gateway® LR reaction
attR1
911 .. 1035  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2491 .. 2615  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2491 .. 2615  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
4816 .. 4937  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4816 .. 4937  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7008 .. 7112  =  105 bp
AmpR promoter
7008 .. 7112  =  105 bp
V5 tag
2641 .. 2682  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
2641 .. 2682  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
2692 .. 2709  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2692 .. 2709  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lac UV5 promoter
1060 .. 1090  =  31 bp
3 segments
   Segment 1:  -35  
   1060 .. 1065  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1060 .. 1090  =  31 bp
3 segments
   Segment 2:  
   1066 .. 1083  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1060 .. 1090  =  31 bp
3 segments
   Segment 3:  -10  
   1084 .. 1090  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1060 .. 1090  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
3630 .. 3765  =  136 bp
SV40 origin of replication
SV40 ori
3630 .. 3765  =  136 bp
SV40 origin of replication
ORF:  1144 .. 1803  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  2959 .. 3225  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  4018 .. 4404  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  6277 .. 6543  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2145 .. 2450  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  3846 .. 4640  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  4155 .. 4409  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  6147 .. 7007  =  861 bp
ORF:  286 amino acids  =  31.6 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
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Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
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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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