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Plasmid Files

patMX4

patMX selector module conferring bialaphos resistance, for gene disruption in yeast, patMX4 version.

 
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 1000 750 500 250 patMX4 TEF promoter BlpR TEF terminator End (1100) BsmI (1087) BglII (890) DraIII (825) BssHII - MauBI (775) BssS α I - EcoNI (754) BmeT110I (739) AvaI - BsoBI (738) BsgI (736) AcuI (697) PasI (693) BtgI - NcoI - StyI (683) BsaI - TaqII (682) AarI (680) BmrI (658) BanII (655) EcoP15I (645) MspA1I (639) BanI (632) AccI (610) SalI (609) NmeAIII (554) BtgZI (540) PfoI * (503) BsiWI (454) BsiHKAI (442) PflFI - Tth111I (415) BglI (397) BsiEI (368) AhdI (364) DraI (294) AflIII - MluI (203) PstI (170) BseRI (150) EaeI (129) Bpu10I (48) Start (0) patMX4 1100 bp
End  (1100)
0 sites
BsmI  (1087)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
BglII  (890)
1 site
A G A T C T T C T A G A
DraIII  (825)
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.
BssHII  (775)
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.
MauBI  (775)
1 site
C G C G C G C G G C G C G C G C
BssSαI  (754)
1 site
C A C G A G G T G C T C
EcoNI  (754)
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.
BmeT110I  (739)
1 site
C Y C G R G G R G C Y C
AvaI  (738)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (738)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
BsgI  (736)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AcuI  (697)
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).
PasI  (693)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
BtgI  (683)
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  (683)
1 site
C C A T G G G G T A C C
StyI  (683)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BsaI  (682)
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.
TaqII  (682)
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.
AarI  (680)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BmrI  (658)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
BanII  (655)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
EcoP15I  (645)
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.
MspA1I  (639)
1 site
C M G C K G G K C G M C
BanI  (632)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
AccI  (610)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
SalI  (609)
1 site
G T C G A C C A G C T G
NmeAIII  (554)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BtgZI  (540)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
PfoI  (503)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
BsiWI  (454)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BsiHKAI  (442)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI sites may not be compatible.
PflFI  (415)
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  (415)
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.
BglI  (397)
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.
BsiEI  (368)
1 site
C G R Y C G G C Y R G C

Sticky ends from different BsiEI sites may not be compatible.
AhdI  (364)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
DraI  (294)
1 site
T T T A A A A A A T T T
AflIII  (203)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (203)
1 site
A C G C G T T G C G C A
PstI  (170)
1 site
C T G C A G G A C G T C
BseRI  (150)
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.
EaeI  (129)
1 site
Y G G C C R R C C G G Y
Bpu10I  (48)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
Start  (0)
0 sites
patMX4
1 .. 1100  =  1100 bp
Product: yeast selectable marker conferring
bialaphos resistance (Goldstein and McCusker,
1999)
patMX4
1 .. 1100  =  1100 bp
Product: yeast selectable marker conferring
bialaphos resistance (Goldstein and McCusker,
1999)
TEF promoter
1 .. 344  =  344 bp
Ashbya gossypii TEF promoter
TEF promoter
1 .. 344  =  344 bp
Ashbya gossypii TEF promoter
BlpR
397 .. 852  =  456 bp
151 amino acids  =  17.1 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialaphos
BlpR
397 .. 852  =  456 bp
151 amino acids  =  17.1 kDa
Product: phosphinothricin acetyltransferase
confers resistance to bialaphos
TEF terminator
903 .. 1100  =  198 bp
Ashbya gossypii TEF terminator
TEF terminator
903 .. 1100  =  198 bp
Ashbya gossypii TEF terminator
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