kanMX4

kanMX selector module conferring kanamycin resistance, for gene disruption in yeast, kanMX4 version.
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1250 1000 750 500 250 End (1429) EcoP15I (1362) ScaI (1194) TatI (1192) PflMI (1068) TaqII (1059) MflI * - BstYI (1054) AseI (1004) BsaWI (940) HindIII (923) Esp3I - BsmBI (821) PvuI - BsiEI - AsiSI (805) BsrFI (759) SspI (730) EcoNI (717) TsoI (646) EarI (618) ClaI - BspDI (496) NruI (462) BanII - Bsp1286I (460) BtgI - StyI - NcoI (378) BciVI (336) DraI (329) MluI - AflIII (238) BfuAI - BspMI (216) PstI (205) BseRI (185) BmgBI (74) BstXI (38) BstEII (21) Start (0) kanMX TEF promoter KanR TEF terminator kanMX4 1429 bp
End  (1429)
0 sites
EcoP15I  (1362)
1 site
C A G C A G ( N ) 25 G T C G T C ( N ) 25 N N

Efficient cleavage requires two inversely oriented copies of the EcoP15I recognition sequence.
Sticky ends from different EcoP15I sites may not be compatible.
EcoP15I requires ATP for activity.
ScaI  (1194)
1 site
A G T A C T T C A T G A
TatI  (1192)
1 site
W G T A C W W C A T G W
PflMI  (1068)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
TaqII  (1059)
1 site
G A C C G A ( N ) 9 N N C T G G C T ( N ) 9

Sticky ends from different TaqII sites may not be compatible.
MflI  (1054)
1 site
R G A T C Y Y C T A G R
* Blocked by Dam methylation.
BstYI  (1054)
1 site
R G A T C Y Y C T A G R
AseI  (1004)
1 site
A T T A A T T A A T T A
BsaWI  (940)
1 site
W C C G G W W G G C C W

Cleavage may be enhanced when more than one copy of the BsaWI recognition sequence is present.
HindIII  (923)
1 site
A A G C T T T T C G A A
Esp3I  (821)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BsmBI  (821)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
PvuI  (805)
1 site
C G A T C G G C T A G C
BsiEI  (805)
1 site
C G R Y C G G C Y R G C

Sticky ends from different BsiEI sites may not be compatible.
AsiSI  (805)
1 site
G C G A T C G C C G C T A G C G
BsrFI  (759)
1 site
R C C G G Y Y G G C C R

Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
SspI  (730)
1 site
A A T A T T T T A T A A
EcoNI  (717)
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.
TsoI  (646)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
EarI  (618)
1 site
C T C T T C N G A G A A G N N N N

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
ClaI  (496)
1 site
A T C G A T T A G C T A
BspDI  (496)
1 site
A T C G A T T A G C T A
NruI  (462)
1 site
T C G C G A A G C G C T
BanII  (460)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
Bsp1286I  (460)
1 site
G D G C H C C H C G D G

Sticky ends from different Bsp1286I sites may not be compatible.
BtgI  (378)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
StyI  (378)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NcoI  (378)
1 site
C C A T G G G G T A C C
BciVI  (336)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
DraI  (329)
1 site
T T T A A A A A A T T T
MluI  (238)
1 site
A C G C G T T G C G C A
AflIII  (238)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
BfuAI  (216)
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  (216)
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.
PstI  (205)
1 site
C T G C A G G A C G T C
BseRI  (185)
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.
BmgBI  (74)
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.
BstXI  (38)
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.
BstEII  (21)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
Start  (0)
0 sites
kanMX
36 .. 1392  =  1357 bp
yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)
kanMX
36 .. 1392  =  1357 bp
yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)
TEF promoter
36 .. 379  =  344 bp
Ashbya gossypii TEF promoter
TEF promoter
36 .. 379  =  344 bp
Ashbya gossypii TEF promoter
KanR
380 .. 1189  =  810 bp
269 amino acids  =  30.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
380 .. 1189  =  810 bp
269 amino acids  =  30.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
TEF terminator
1195 .. 1392  =  198 bp
Ashbya gossypii TEF terminator
TEF terminator
1195 .. 1392  =  198 bp
Ashbya gossypii TEF terminator
ORF:  1 .. 246  =  246 bp
ORF:  81 amino acids  =  9.3 kDa  (no start codon)
ORF:  380 .. 1189  =  810 bp
ORF:  269 amino acids  =  30.7 kDa
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Download kanMX4.dna file

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