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Plasmid Files

pDONR™/Zeo

Gateway® donor vector with attP1 and attP2 sites and a Zeocin™ resistance marker.

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pDONR Zeo Sequence and MappDONR Zeo.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 PciI (4285) DrdI (4183) BssS α I (4112) PspFI (3985) BseYI (3981) AcuI (3743) PmlI (3553) MauBI (3444) AatII (3442) ZraI (3440) SgrAI (3366) SexAI * (3316) BanI (3310) FseI (3212) BglI (3182) EagI (3149) DraIII (3134) BseRI (3111) EcoRV (2999) BspEI (2292) ApoI - EcoRI (2288) NspI (4289) BspQI - SapI (111) Bts α I (214) BbsI (437) HpaI (501) M13 fwd AflII (554) PspOMI (563) EcoO109I (564) ApaI - BanII (567) XmnI (1052) PstI (1167) BfuAI - BspMI (1170) BsaI (1276) BstXI (1305) BsaBI * (1405) SrfI (1418) BbvCI (1561) BstZ17I (1754) BamHI (1832) ScaI (1875) SspI (1980) BpmI (2172) pDONR™/Zeo 4291 bp
PciI  (4285)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (4183)
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.
BssSαI  (4112)
1 site
C A C G A G G T G C T C
PspFI  (3985)
1 site
C C C A G C G G G T C G
BseYI  (3981)
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  (3743)
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).
PmlI  (3553)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
MauBI  (3444)
1 site
C G C G C G C G G C G C G C G C
AatII  (3442)
1 site
G A C G T C C T G C A G
ZraI  (3440)
1 site
G A C G T C C T G C A G
SgrAI  (3366)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
SexAI  (3316)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BanI  (3310)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
FseI  (3212)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
BglI  (3182)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
EagI  (3149)
1 site
C G G C C G G C C G G C
DraIII  (3134)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BseRI  (3111)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
EcoRV  (2999)
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.
BspEI  (2292)
1 site
T C C G G A A G G C C T
ApoI  (2288)
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  (2288)
1 site
G A A T T C C T T A A G
NspI  (4289)
1 site
R C A T G Y Y G T A C R
BspQI  (111)
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  (111)
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.
BtsαI  (214)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
BbsI  (437)
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  (501)
1 site
G T T A A C C A A T T G
AflII  (554)
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  (563)
1 site
G G G C C C C C C G G G
EcoO109I  (564)
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  (567)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (567)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
XmnI  (1052)
1 site
G A A N N N N T T C C T T N N N N A A G
PstI  (1167)
1 site
C T G C A G G A C G T C
BfuAI  (1170)
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  (1170)
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  (1276)
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  (1305)
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.
BsaBI  (1405)
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  (1418)
1 site
G C C C G G G C C G G G C C C G
BbvCI  (1561)
1 site
C C T C A G C G G A G T C G
BstZ17I  (1754)
1 site
G T A T A C C A T A T G
BamHI  (1832)
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  (1875)
1 site
A G T A C T T C A T G A
SspI  (1980)
1 site
A A T A T T T T A T A A
BpmI  (2172)
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.
CmR
1847 .. 2506  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1847 .. 2506  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3641 .. 4229  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3641 .. 4229  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
BleoR
3111 .. 3485  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
BleoR
3111 .. 3485  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
ccdB
1197 .. 1502  =  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
1197 .. 1502  =  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
570 .. 801  =  232 bp
recombination site for the Gateway® BP reaction
(pDONR™221 version)
attP1
570 .. 801  =  232 bp
recombination site for the Gateway® BP reaction
(pDONR™221 version)
attP2
2754 .. 2985  =  232 bp
recombination site for the Gateway® BP reaction
(pDONR™221 version)
attP2
2754 .. 2985  =  232 bp
recombination site for the Gateway® BP reaction
(pDONR™221 version)
cat promoter
2507 .. 2609  =  103 bp
promoter of the E. coli cat gene
cat promoter
2507 .. 2609  =  103 bp
promoter of the E. coli cat gene
rrnB T1 terminator
387 .. 473  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
387 .. 473  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
EM7 promoter
3504 .. 3551  =  48 bp
synthetic bacterial promoter
EM7 promoter
3504 .. 3551  =  48 bp
synthetic bacterial promoter
rrnB T2 terminator
268 .. 295  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
268 .. 295  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
M13 fwd
537 .. 553  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
537 .. 553  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
3027 .. 3043  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
3027 .. 3043  =  17 bp
common sequencing primer, one of multiple similar
variants
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