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Plasmid Files

natMX6

natMX6 selector module conferring nourseothricin resistance, for gene disruption in yeast.

 
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natMX6.dna
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1000 750 500 250 natMX6 TEF promoter NrsR TEF terminator End (1123) BsmI (1110) ScaI (925) TatI (923) BstAPI (910) NspI - SphI (908) EciI (833) BpmI - Eco57MI (795) HincII (791) BstEII (779) PluTI (760) SfoI (758) NarI (757) KasI (756) MreI (744) NruI (732) BanII (730) BssHII (713) BsiHKAI (694) DrdI (678) BsiWI (595) PshAI (588) PfoI (579) PflFI - Tth111I (444) BbsI (440) EcoO109I - PpuMI (427) XcmI (416) BglI (414) SmaI (394) BmeT110I (393) AvaI - BsoBI - TspMI - XmaI (392) AgeI - BsaWI (373) BtgI - NcoI (343) BciVI (301) DraI (294) BfuAI - BspMI (181) PstI (170) BseRI (150) EaeI (129) Bpu10I (48) BmgBI (39) Start (0) natMX6 1123 bp
End  (1123)
0 sites
BsmI  (1110)
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.
ScaI  (925)
1 site
A G T A C T T C A T G A
TatI  (923)
1 site
W G T A C W W C A T G W
BstAPI  (910)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
NspI  (908)
1 site
R C A T G Y Y G T A C R
SphI  (908)
1 site
G C A T G C C G T A C G
EciI  (833)
1 site
G G C G G A ( N ) 9 N N C C G C C T ( N ) 9

Sticky ends from different EciI sites may not be compatible.
BpmI  (795)
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.
Eco57MI  (795)
1 site
C T G R A G ( N ) 14 N N G A C Y T C ( N ) 14

Sticky ends from different Eco57MI sites may not be compatible.
After cleavage, Eco57MI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
HincII  (791)
1 site
G T Y R A C C A R Y T G
BstEII  (779)
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.
PluTI  (760)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (758)
1 site
G G C G C C C C G C G G
NarI  (757)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (756)
1 site
G G C G C C C C G C G G
MreI  (744)
1 site
C G C C G G C G G C G G C C G C
NruI  (732)
1 site
T C G C G A A G C G C T
BanII  (730)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BssHII  (713)
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.
BsiHKAI  (694)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI sites may not be compatible.
DrdI  (678)
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.
BsiWI  (595)
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.
PshAI  (588)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
PfoI  (579)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
PflFI  (444)
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  (444)
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.
BbsI  (440)
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.
EcoO109I  (427)
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.
PpuMI  (427)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
XcmI  (416)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BglI  (414)
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.
SmaI  (394)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BmeT110I  (393)
1 site
C Y C G R G G R G C Y C
AvaI  (392)
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  (392)
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.
TspMI  (392)
1 site
C C C G G G G G G C C C
XmaI  (392)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
AgeI  (373)
1 site
A C C G G T T G G C C A
BsaWI  (373)
1 site
W C C G G W W G G C C W

Efficient cleavage requires at least two copies of the BsaWI recognition sequence.
BtgI  (343)
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  (343)
1 site
C C A T G G G G T A C C
BciVI  (301)
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  (294)
1 site
T T T A A A A A A T T T
BfuAI  (181)
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  (181)
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  (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.
BmgBI  (39)
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.
Start  (0)
0 sites
natMX6
1 .. 1123  =  1123 bp
yeast selectable marker conferring nourseothricin resistance
natMX6
1 .. 1123  =  1123 bp
yeast selectable marker conferring nourseothricin resistance
TEF promoter
1 .. 344  =  344 bp
Ashbya gossypii TEF promoter
TEF promoter
1 .. 344  =  344 bp
Ashbya gossypii TEF promoter
NrsR
345 .. 920  =  576 bp
191 amino acids  =  20.6 kDa
Product: nourseothricin acetyltransferase
confers resistance to nourseothricin
NrsR
345 .. 920  =  576 bp
191 amino acids  =  20.6 kDa
Product: nourseothricin acetyltransferase
confers resistance to nourseothricin
TEF terminator
926 .. 1123  =  198 bp
Ashbya gossypii TEF terminator
TEF terminator
926 .. 1123  =  198 bp
Ashbya gossypii TEF terminator
ORF:  345 .. 920  =  576 bp
ORF:  191 amino acids  =  20.6 kDa
ORF:  371 .. 676  =  306 bp
ORF:  101 amino acids  =  10.5 kDa
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