Resources
Plasmid Files

pDONR™222

Gateway® donor vector with attP1 and attP2 sites and a kanamycin resistance marker. The alternative pDONR™221 vector seems to be preferred.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pDONR222 Sequence and MappDONR222.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
Download Free Trial Get SnapGene Viewer

 AcuI (4591) PspFI (4351) BseYI (4347) BssS α I (4216) DrdI (4151) NspI (4047) PciI (4043) BspQI - SapI (3927) BglII (3715) PaeR7I - PspXI - XhoI (3680) NruI (3625) BspDI - ClaI (3589) EcoNI (3369) AsiSI - PvuI (3284) HindIII (3160) EcoRV (2788) BsaAI (2388) BsaHI (117) BbsI (227) HpaI (291) M13 fwd AflII (344) PspOMI (353) EcoO109I (354) ApaI (357) XmnI (842) PstI (957) BfuAI - BspMI (960) BsaI (1066) BstXI (1095) BmgBI (1174) BsaBI * (1195) SrfI (1208) BbvCI (1351) BstZ17I (1544) BssHII (1581) BamHI (1622) ScaI (1664) BtgI - NcoI - StyI (1776) BpmI (1961) EcoRI (2077) BspEI (2081) pDONR™222 4718 bp
AcuI  (4591)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI
recognition sequence.
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).
PspFI  (4351)
1 site
C C C A G C G G G T C G
BseYI  (4347)
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.
BssSαI  (4216)
1 site
C A C G A G G T G C T C
DrdI  (4151)
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.
NspI  (4047)
1 site
R C A T G Y Y G T A C R
PciI  (4043)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3927)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (3927)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
BglII  (3715)
1 site
A G A T C T T C T A G A
PaeR7I  (3680)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3680)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3680)
1 site
C T C G A G G A G C T C
NruI  (3625)
1 site
T C G C G A A G C G C T
BspDI  (3589)
1 site
A T C G A T T A G C T A
ClaI  (3589)
1 site
A T C G A T T A G C T A
EcoNI  (3369)
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.
AsiSI  (3284)
1 site
G C G A T C G C C G C T A G C G
PvuI  (3284)
1 site
C G A T C G G C T A G C
HindIII  (3160)
1 site
A A G C T T T T C G A A
EcoRV  (2788)
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.
BsaAI  (2388)
1 site
Y A C G T R R T G C A Y
BsaHI  (117)
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.
BbsI  (227)
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  (291)
1 site
G T T A A C C A A T T G
AflII  (344)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
PspOMI  (353)
1 site
G G G C C C C C C G G G
EcoO109I  (354)
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  (357)
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  (842)
1 site
G A A N N N N T T C C T T N N N N A A G
PstI  (957)
1 site
C T G C A G G A C G T C
BfuAI  (960)
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  (960)
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  (1066)
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  (1095)
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  (1174)
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  (1195)
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.
SrfI  (1208)
1 site
G C C C G G G C C G G G C C C G
BbvCI  (1351)
1 site
C C T C A G C G G A G T C G
BstZ17I  (1544)
1 site
G T A T A C C A T A T G
BssHII  (1581)
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  (1622)
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  (1664)
1 site
A G T A C T T C A T G A
BtgI  (1776)
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  (1776)
1 site
C C A T G G G G T A C C
StyI  (1776)
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  (1961)
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  (2077)
1 site
G A A T T C C T T A A G
BspEI  (2081)
1 site
T C C G G A A G G C C T
KanR
2899 .. 3714  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
2899 .. 3714  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
CmR
1642 .. 2295  =  654 bp
218 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1642 .. 2295  =  654 bp
218 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
4104 .. 4692  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4104 .. 4692  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ccdB
987 .. 1292  =  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
987 .. 1292  =  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.
attP1
360 .. 591  =  232 bp
recombination site for the Gateway® BP reaction
(pDONR™201 version)
attP1
360 .. 591  =  232 bp
recombination site for the Gateway® BP reaction
(pDONR™201 version)
attP2
2543 .. 2774  =  232 bp
recombination site for the Gateway® BP reaction
(pDONR™201 version)
attP2
2543 .. 2774  =  232 bp
recombination site for the Gateway® BP reaction
(pDONR™201 version)
cat promoter
2296 .. 2398  =  103 bp
promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase
cat promoter
2296 .. 2398  =  103 bp
promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase
rrnB T1 terminator
177 .. 263  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
177 .. 263  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T2 terminator
58 .. 85  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
58 .. 85  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
M13 fwd
327 .. 343  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
327 .. 343  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2816 .. 2832  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2816 .. 2832  =  17 bp
common sequencing primer, one of multiple similar
variants
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter