pDONR222

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

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspDONR222
AcuI (4591) PspFI (4351) BseYI (4347) BssSI - BssSαI (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) rrnB T2 terminator BsaHI (117) BbsI (227) HpaI (291) 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
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).
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.
BssSI  (4216)
1 site		 C A C G A G G T G C T C
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
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
ORF:  987 .. 1292  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4438 .. 67  =  348 bp
ORF:  115 amino acids  =  12.9 kDa
ORF:  1612 .. 2295  =  684 bp
ORF:  227 amino acids  =  26.7 kDa
ORF:  2899 .. 3759  =  861 bp
ORF:  286 amino acids  =  32.8 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

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