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Plasmid Files

bleMX6

bleMX6 selector module conferring phleomycin resistance, for gene disruption in yeast.

 
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bleMX6.dna
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800 600 400 200 bleMX6 TEF promoter BleoR TEF terminator End (922) BsmI (909) ScaI (724) TatI (722) BclI * (717) DraIII (698) BaeGI - Bme1580I (694) ApaLI (690) EagI (676) BglI (650) FseI (621) MmeI (559) BanI (515) SexAI * (508) SgrAI (459) SmaI (435) TspMI - XmaI (433) BsrBI (399) AatII (391) ZraI (389) BssHII - MauBI (381) BsaWI (368) HincII (355) MscI (348) NcoI - StyI (343) BciVI (301) DraI (294) AflIII - MluI (203) BfuAI - BspMI (181) PstI (170) Bpu10I (48) Start (0) bleMX6 922 bp
End  (922)
0 sites
BsmI  (909)
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  (724)
1 site
A G T A C T T C A T G A
TatI  (722)
1 site
W G T A C W W C A T G W
BclI  (717)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
DraIII  (698)
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.
BaeGI  (694)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (694)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
ApaLI  (690)
1 site
G T G C A C C A C G T G
EagI  (676)
1 site
C G G C C G G C C G G C
BglI  (650)
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.
FseI  (621)
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.
MmeI  (559)
1 site
T C C R A C ( N ) 18 N N A G G Y T G ( N ) 18

Efficient cleavage requires at least two copies of the MmeI recognition sequence.
Sticky ends from different MmeI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BanI  (515)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
SexAI  (508)
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.
SgrAI  (459)
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.
SmaI  (435)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (433)
1 site
C C C G G G G G G C C C
XmaI  (433)
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.
BsrBI  (399)
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.
AatII  (391)
1 site
G A C G T C C T G C A G
ZraI  (389)
1 site
G A C G T C C T G C A G
BssHII  (381)
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  (381)
1 site
C G C G C G C G G C G C G C G C
BsaWI  (368)
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.
HincII  (355)
1 site
G T Y R A C C A R Y T G
MscI  (348)
1 site
T G G C C A A C C G G T
NcoI  (343)
1 site
C C A T G G G G T A C C
StyI  (343)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
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
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
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
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
bleMX6
1 .. 922  =  922 bp
yeast selectable marker conferring phleomycin resistance
bleMX6
1 .. 922  =  922 bp
yeast selectable marker conferring phleomycin resistance
TEF promoter
1 .. 344  =  344 bp
Ashbya gossypii TEF promoter
TEF promoter
1 .. 344  =  344 bp
Ashbya gossypii TEF promoter
BleoR
345 .. 719  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
345 .. 719  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
TEF terminator
725 .. 922  =  198 bp
Ashbya gossypii TEF terminator
TEF terminator
725 .. 922  =  198 bp
Ashbya gossypii TEF terminator
ORF:  345 .. 719  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  323 .. 625  =  303 bp
ORF:  100 amino acids  =  10.8 kDa
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