pDEST R4-R3 Vector II

Improved Gateway® destination vector for generating an expression clone by three-fragment multisite recombination.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspDEST R4-R3 Vector II
AflIII - PciI (4466) PspFI (4166) BseYI (4162) AhdI (3578) NmeAIII (3431) AseI (3403) BsrBI (2736) AatII (2659) ZraI (2657) EcoO109I (2598) PfoI (2539) BstAPI (2408) NdeI (2404) PluTI (2355) SfoI (2353) NarI (2352) KasI (2351) SalI (161) PstI (478) BfuAI - BspMI (481) BstXI (616) BmgBI (695) BsaBI * (716) AvaI - BsoBI - TspMI - XmaI (727) SmaI - SrfI (729) BsrGI (752) BbvCI (872) BstZ17I (1065) BssHII (1102) BamHI (1143) BtgI - NcoI - StyI (1297) ApoI - EcoRI (1598) BspEI (1602) BsaAI (1909) EagI (2058) M13 fwd pDEST™ R4-R3 Vector II 4555 bp
AflIII  (4466)
1 site		 A C R Y G T T G Y R C A
Sticky ends from different AflIII sites may not be compatible.
PciI  (4466)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
PspFI  (4166)
1 site		 C C C A G C G G G T C G
BseYI  (4162)
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.
AhdI  (3578)
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.
NmeAIII  (3431)
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).
AseI  (3403)
1 site		 A T T A A T T A A T T A
BsrBI  (2736)
1 site		 C C G C T C G G C G A G
This recognition sequence is asymmetric, so ligating blunt ends generated by BsrBI will not always regenerate a BsrBI site.
BsrBI is typically used at 37°C, but can be used at temperatures up to 50°C.
AatII  (2659)
1 site		 G A C G T C C T G C A G
ZraI  (2657)
1 site		 G A C G T C C T G C A G
EcoO109I  (2598)
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.
PfoI  (2539)
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  (2408)
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.
NdeI  (2404)
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.
PluTI  (2355)
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  (2353)
1 site		 G G C G C C C C G C G G
NarI  (2352)
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  (2351)
1 site		 G G C G C C C C G C G G
SalI  (161)
1 site		 G T C G A C C A G C T G
PstI  (478)
1 site		 C T G C A G G A C G T C
BfuAI  (481)
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  (481)
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.
BstXI  (616)
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.
BmgBI  (695)
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.
BsaBI  (716)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
AvaI  (727)
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  (727)
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  (727)
1 site		 C C C G G G G G G C C C
XmaI  (727)
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  (729)
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  (729)
1 site		 G C C C G G G C C G G G C C C G
BsrGI  (752)
1 site		 T G T A C A A C A T G T
BsrGI is typically used at 37°C, but is even more active at 60°C.
BbvCI  (872)
1 site		 C C T C A G C G G A G T C G
BstZ17I  (1065)
1 site		 G T A T A C C A T A T G
BssHII  (1102)
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.
BamHI  (1143)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BtgI  (1297)
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  (1297)
1 site		 C C A T G G G G T A C C
StyI  (1297)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
ApoI  (1598)
1 site		 R A A T T Y Y T T A A R
ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (1598)
1 site		 G A A T T C C T T A A G
BspEI  (1602)
1 site		 T C C G G A A G G C C T
BsaAI  (1909)
1 site		 Y A C G T R R T G C A Y
EagI  (2058)
1 site		 C G G C C G G C C G G C
AmpR
2791 .. 3651  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   2791 .. 2859  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2791 .. 3651  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   2860 .. 3651  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2791 .. 3651  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
1163 .. 1816  =  654 bp
218 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1163 .. 1816  =  654 bp
218 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3822 .. 4410  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3822 .. 4410  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ccdB
508 .. 813  =  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
508 .. 813  =  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.
attR4
36 .. 160  =  125 bp
recombination site for the Gateway® LR reaction
attR4
36 .. 160  =  125 bp
recombination site for the Gateway® LR reaction
attR3
2065 .. 2188  =  124 bp
recombination site for the Gateway® LR reaction
attR3
2065 .. 2188  =  124 bp
recombination site for the Gateway® LR reaction
AmpR promoter
2686 .. 2790  =  105 bp
AmpR promoter
2686 .. 2790  =  105 bp
cat promoter
1817 .. 1919  =  103 bp
promoter of the E. coli cat gene
cat promoter
1817 .. 1919  =  103 bp
promoter of the E. coli cat gene
M13 rev
1 .. 17  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
1 .. 17  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2196 .. 2212  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2196 .. 2212  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  2791 .. 3651  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1133 .. 1816  =  684 bp
ORF:  227 amino acids  =  26.7 kDa
ORF:  508 .. 813  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  2154 .. 2405  =  252 bp
ORF:  83 amino acids  =  9.8 kDa
ORF:  3255 .. 3521  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Download pDEST R4-R3 Vector II.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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