pDONR207

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

Sequence Author: Thermo Fisher (Invitrogen)

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DrdI (5477) BssSI - BssSαI (5406) PspFI (5279) BseYI (5275) BglI (4492) EagI (4484) DraIII (4309) PflFI - Tth111I (3954) BglII (3743) BseRI (3687) EcoRV (3551) AsiSI - PvuI (3293) EcoNI (3205) NruI (2950) BsaHI (132) BsrBI (181) HpaI (306) PspOMI (325) EcoO109I (326) ApaI (329) BfuAI - BspMI (932) BstXI (1067) BmgBI (1146) TspMI - XmaI (1178) SmaI - SrfI (1180) BbvCI (1323) BstZ17I (1516) BssHII (1553) BamHI (1594) ScaI (1634) NcoI (1746) BpmI (1931) EcoRI (2047) BspEI (2051) pDONR™207 5585 bp
DrdI  (5477)
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  (5406)
1 site
C A C G A G G T G C T C
BssSαI  (5406)
1 site
C A C G A G G T G C T C
PspFI  (5279)
1 site
C C C A G C G G G T C G
BseYI  (5275)
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.
BglI  (4492)
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  (4484)
1 site
C G G C C G G C C G G C
DraIII  (4309)
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.
PflFI  (3954)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3954)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BglII  (3743)
1 site
A G A T C T T C T A G A
BseRI  (3687)
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  (3551)
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.
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
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.
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.
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.
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.
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
NcoI  (1746)
1 site
C C A T G G G G T A C C
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
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
4935 .. 5523  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4935 .. 5523  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
GmR
3528 .. 4061  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
3528 .. 4061  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
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 propagated 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 propagated 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:  4163 .. 4792  =  630 bp
ORF:  209 amino acids  =  24.1 kDa
ORF:  2868 .. 3998  =  1131 bp
ORF:  376 amino acids  =  42.0 kDa
ORF:  4272 .. 4514  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  3528 .. 4061  =  534 bp
ORF:  177 amino acids  =  19.4 kDa
ORF:  959 .. 1264  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4253 .. 4561  =  309 bp
ORF:  102 amino acids  =  11.4 kDa
ORF:  1606 .. 2265  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
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