pENTR2B

Gateway® Dual Selection Vector for creating an entry vector by restriction cloning. Other reading frames are available with pENTR™1A and pENTR™3C.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspENTR2B
NspI (3753) AflIII - PciI (3749) DrdI (3647) BssSI - BssSαI (3576) BciVI (3551) AcuI (3207) TaqII (2885) AseI (2830) AsiSI - PvuI (2631) EcoNI (2543) NruI (2288) BspHI (2155) EcoRV (1977) PaeR7I - XhoI (1970) BfuAI - BspMI (1936) NheI (44) BmtI (48) ZraI (64) AatII (66) BbsI (134) BmrI (141) BsrBI (199) AflII (343) KasI (458) NarI (459) SfoI (460) PluTI (462) XmnI (469) Acc65I (489) KpnI (493) BspEI (818) BpmI (944) PasI (1054) BtgI - NcoI - StyI (1123) ScaI (1239) BssHII (1315) BstZ17I (1356) BbvCI (1546) TspMI - XmaI (1690) SmaI - SrfI (1692) BsaBI * (1705) BmgBI (1726) BstXI (1809) BsaI (1830) pENTR™2B 3755 bp
NspI  (3753)
1 site		 R C A T G Y Y G T A C R
AflIII  (3749)
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  (3749)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
DrdI  (3647)
1 site		 G A C N N N N N N G T C C T G N N N N N N C A G
Sticky ends from different DrdI sites may not be compatible.
BssSI  (3576)
1 site		 C A C G A G G T G C T C
BssSαI  (3576)
1 site		 C A C G A G G T G C T C
BciVI  (3551)
1 site		 G T A T C C ( N ) 5 N C A T A G G ( N ) 5
The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
AcuI  (3207)
1 site		 C T G A A G ( N ) 14 N N G A C T T C ( N ) 14
Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
TaqII  (2885)
1 site		 G A C C G A ( N ) 9 N N C T G G C T ( N ) 9
Sticky ends from different TaqII sites may not be compatible.
AseI  (2830)
1 site		 A T T A A T T A A T T A
AsiSI  (2631)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (2631)
1 site		 C G A T C G G C T A G C
EcoNI  (2543)
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.
NruI  (2288)
1 site		 T C G C G A A G C G C T
BspHI  (2155)
1 site		 T C A T G A A G T A C T
EcoRV  (1977)
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.
PaeR7I  (1970)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1970)
1 site		 C T C G A G G A G C T C
BfuAI  (1936)
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  (1936)
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.
NheI  (44)
1 site		 G C T A G C C G A T C G
BmtI  (48)
1 site		 G C T A G C C G A T C G
ZraI  (64)
1 site		 G A C G T C C T G C A G
AatII  (66)
1 site		 G A C G T C C T G C A G
BbsI  (134)
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.
BmrI  (141)
1 site		 A C T G G G ( N ) 4 N T G A C C C ( N ) 4
The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
BsrBI  (199)
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.
AflII  (343)
1 site		 C T T A A G G A A T T C
KasI  (458)
1 site		 G G C G C C C C G C G G
NarI  (459)
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  (460)
1 site		 G G C G C C C C G C G G
PluTI  (462)
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.
XmnI  (469)
1 site		 G A A N N N N T T C C T T N N N N A A G
Acc65I  (489)
1 site		 G G T A C C C C A T G G
KpnI  (493)
1 site		 G G T A C C C C A T G G
BspEI  (818)
1 site		 T C C G G A A G G C C T
BpmI  (944)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
PasI  (1054)
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.
BtgI  (1123)
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  (1123)
1 site		 C C A T G G G G T A C C
StyI  (1123)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
ScaI  (1239)
1 site		 A G T A C T T C A T G A
BssHII  (1315)
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  (1356)
1 site		 G T A T A C C A T A T G
BbvCI  (1546)
1 site		 C C T C A G C G G A G T C G
TspMI  (1690)
1 site		 C C C G G G G G G C C C
XmaI  (1690)
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  (1692)
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  (1692)
1 site		 G C C C G G G C C G G G C C C G
BsaBI  (1705)
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.
BmgBI  (1726)
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  (1809)
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.
BsaI  (1830)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
KanR
2206 .. 3015  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
2206 .. 3015  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
CmR
609 .. 1289  =  681 bp
226 amino acids  =  26.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
609 .. 1289  =  681 bp
226 amino acids  =  26.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3105 .. 3693  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3105 .. 3693  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ccdB
1609 .. 1914  =  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
1609 .. 1914  =  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.
attL1
358 .. 457  =  100 bp
recombination site for the Gateway® LR reaction
attL1
358 .. 457  =  100 bp
recombination site for the Gateway® LR reaction
attL2
1984 .. 2083  =  100 bp
recombination site for the Gateway® LR reaction
attL2
1984 .. 2083  =  100 bp
recombination site for the Gateway® LR reaction
rrnB T1 terminator
103 .. 189  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
103 .. 189  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
MCS 1
458 .. 508  =  51 bp
multiple cloning site, part 1
MCS 1
458 .. 508  =  51 bp
multiple cloning site, part 1
MCS 2
1949 .. 1980  =  32 bp
multiple cloning site, part 2
MCS 2
1949 .. 1980  =  32 bp
multiple cloning site, part 2
lac UV5 promoter
525 .. 555  =  31 bp
3 segments
   Segment 1:  -35  
   525 .. 530  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
525 .. 555  =  31 bp
3 segments
   Segment 2:  
   531 .. 548  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
525 .. 555  =  31 bp
3 segments
   Segment 3:  -10  
   549 .. 555  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
525 .. 555  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
rrnB T2 terminator
281 .. 308  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
281 .. 308  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
ORF:  1609 .. 1914  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  2206 .. 3015  =  810 bp
ORF:  269 amino acids  =  30.8 kDa
ORF:  609 .. 1289  =  681 bp
ORF:  226 amino acids  =  26.6 kDa
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Download pENTR2B.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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