pDONR201

Donor vector for inserting an attB-flanked gene to generate an attL-containing entry vector in the Gateway® system.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsI.M.A.G.E. Consortium PlasmidspDONR201
AflIII - PciI (4464) DrdI (4362) BssSI - BssSαI (4291) HaeII (4224) PspFI (4164) BseYI (4160) AcuI (3922) AseI (3492) AsiSI - PvuI (3293) EcoNI (3205) NruI (2950) BsaAI (2358) NspI (4468) BsaHI (132) BsrBI (181) BmrI (240) BbsI (242) HpaI (306) PspOMI (325) EcoO109I (326) ApaI (329) XmnI (814) BfuAI - BspMI (932) BsaI (1038) BstXI (1067) BmgBI (1146) BsaBI * (1167) TspMI - XmaI (1178) SmaI - SrfI (1180) BbvCI (1323) BstZ17I (1516) BssHII (1553) BamHI (1594) ScaI (1634) BtgI - NcoI - StyI (1746) BpmI (1931) EcoRI (2047) BspEI (2051) pDONR™201 4470 bp
AflIII  (4464)
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  (4464)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
DrdI  (4362)
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  (4291)
1 site		 C A C G A G G T G C T C
BssSαI  (4291)
1 site		 C A C G A G G T G C T C
HaeII  (4224)
1 site		 R G C G C Y Y C G C G R
PspFI  (4164)
1 site		 C C C A G C G G G T C G
BseYI  (4160)
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.
AcuI  (3922)
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).
AseI  (3492)
1 site		 A T T A A T T A A T T A
AsiSI  (3293)
1 site		 G C G A T C G C C G C T A G C G
PvuI  (3293)
1 site		 C G A T C G G C T A G C
EcoNI  (3205)
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  (2950)
1 site		 T C G C G A A G C G C T
BsaAI  (2358)
1 site		 Y A C G T R R T G C A Y
NspI  (4468)
1 site		 R C A T G Y Y G T A C R
BsaHI  (132)
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.
BsrBI  (181)
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.
BmrI  (240)
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.
BbsI  (242)
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.
HpaI  (306)
1 site		 G T T A A C C A A T T G
PspOMI  (325)
1 site		 G G G C C C C C C G G G
EcoO109I  (326)
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.
ApaI  (329)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
XmnI  (814)
1 site		 G A A N N N N T T C C T T N N N N A A G
BfuAI  (932)
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  (932)
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.
BsaI  (1038)
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.
BstXI  (1067)
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  (1146)
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  (1167)
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.
TspMI  (1178)
1 site		 C C C G G G G G G C C C
XmaI  (1178)
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  (1180)
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  (1180)
1 site		 G C C C G G G C C G G G C C C G
BbvCI  (1323)
1 site		 C C T C A G C G G A G T C G
BstZ17I  (1516)
1 site		 G T A T A C C A T A T G
BssHII  (1553)
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  (1594)
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.
ScaI  (1634)
1 site		 A G T A C T T C A T G A
BtgI  (1746)
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  (1746)
1 site		 C C A T G G G G T A C C
StyI  (1746)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
BpmI  (1931)
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.
EcoRI  (2047)
1 site		 G A A T T C C T T A A G
BspEI  (2051)
1 site		 T C C G G A A G G C C T
KanR
2868 .. 3677  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
2868 .. 3677  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
CmR
1606 .. 2265  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1606 .. 2265  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3820 .. 4408  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3820 .. 4408  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ccdB
959 .. 1264  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagaged in standard E. coli strains.
ccdB
959 .. 1264  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagaged in standard E. coli strains.
attP1
332 .. 563  =  232 bp
recombination site for the Gateway® BP reaction (pDONR™201 version)
attP1
332 .. 563  =  232 bp
recombination site for the Gateway® BP reaction (pDONR™201 version)
attP2
2513 .. 2744  =  232 bp
recombination site for the Gateway® BP reaction (pDONR™201 version)
attP2
2513 .. 2744  =  232 bp
recombination site for the Gateway® BP reaction (pDONR™201 version)
cat promoter
2266 .. 2368  =  103 bp
promoter of the E. coli cat gene
cat promoter
2266 .. 2368  =  103 bp
promoter of the E. coli cat gene
rrnB T1 terminator
192 .. 278  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
192 .. 278  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T2 terminator
73 .. 100  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
73 .. 100  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
ORF:  2868 .. 3677  =  810 bp
ORF:  269 amino acids  =  30.8 kDa
ORF:  1606 .. 2265  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  959 .. 1264  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
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Individual Sequences & Maps

Lafmid BA
pcDNAII
pCR2.1-TOPO (linearized)
pExpress-1
pAD-GAL4
pcDNA3.1(+)
pCR3.1
pME18S-FL3
pAMP1
pcDNA3.1(-)
pCR3.1 (linearized)
pOTB7
pAMP10
pcDNA3.1 Zeo(+)
pCR4-TOPO
pOTB7-3
pBK-CMV
pCMV SPORT
pCR4-TOPO (linearized)
pOTB7a
pBluescribe (modified)
pCMV SPORT2
pCS105
pPCR-Script Amp SK(+)
pBluescript II SK(+)
pCMV SPORT4
pCS107
pRKW2
pBluescript KS(+)
pCMV SPORT6
pCS108
pSPORT 1
pBluescript SK(+)
pCMV SPORT6.1
pCS2+
pSPORT 2
pBluescript SK(-)
pCMV SPORT6ccdB
pCS22+
pT7T3D-PacI
pBluescript-FL
pCR-Blunt II-TOPO
pCS2G
pUC19
pBluescript II KS(+)
pCR-Blunt II-TOPO (linearized)
pDNR-Dual
pYX-Asc
pBluescriptR
pCR-XL-TOPO
pDNR-LIB
pZErO-2
pcDNA3
pCR-XL-TOPO (linearized)
pDONR201
pZL1
pcDNAI
pCR2.1-TOPO
pDONR223

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