bleMX6

bleMX6 selector module conferring phleomycin resistance, for gene disruption in yeast.
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HomePlasmidsYeast PlasmidsbleMX6
800 600 400 200 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) CsiI - SexAI * (508) SgrAI (459) SmaI (435) XmaI - TspMI (433) BsrBI (399) AatII (391) ZraI (389) MauBI - BssHII (381) BsaWI (368) HincII (355) MscI (348) StyI NcoI (343) BciVI (301) DraI (294) MluI - AflIII (203) BfuAI - BspMI (181) PstI (170) Bpu10I (48) Start (0) bleMX6 TEF promoter BleoR TEF terminator 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.
CsiI  (508)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI 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.
XmaI  (433)
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.
TspMI  (433)
1 site		 C C C G G G G G G C C C
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
MauBI  (381)
1 site		 C G C G C G C G G C G C G C G C
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.
BsaWI  (368)
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.
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
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.
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
MluI  (203)
1 site		 A C G C G T T G C G C A
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.
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
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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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Download bleMX6.dna file

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Individual Sequences & Maps

bleMX6
pFA6a-kanMX6
pPIC6(alpha) B
pYES2.1-E
hphMX6
pFA6a-kanMX6-PGAL1
pPIC6(alpha) C
pYES2 CT
kanMX
pFA6a-kanMX6-PGAL1-3HA
pPIC9
pYES2 NT A
kanMX2
pFA6a-kanMX6-PGAL1-GFP
pPIC9K
pYES2 NT B
kanMX4
pFA6a-kanMX6-PGAL1-GST
pPICZ A
pYES2 NT C
kanMX6
pFA6a-natMX6
pPICZ B
pYES3 CT
natMX6
pFA6a-TRP1
pPICZ C
pYES6 CT
pAbAi
pFA6a-TRP1-PGAL1
pPICZ(alpha) A
pYM25
pAD-GAL4
pFA6a-TRP1-PGAL1-3HA
pPICZ(alpha) B
pYM26
pAG25
pFA6a-TRP1-PGAL1-GFP
pPICZ(alpha) C
pYM44
pAG29
pFA6a-TRP1-PGAL1-GST
pPink-HC
YCp50
pAG32
pFLD
pPink-LC
YCp50-poly
pAO815
pFLD(alpha)
pPINK(alpha)-HC
YCplac111
patMX4
pGAD-C1
pRS303
YCplac22
pAUR101
pGAD-C2
pRS304
YCplac33
pAUR112
pGAD-C3
pRS305
YEp13
pAUR123
pGADT7 AD
pRS306
YEp24
pAUR224
pGADT7-Rec
pRS313
YEp351
pBridge
pGADT7-Rec (linearized)
pRS314
YEp352
pDH25
pGAPZ A
pRS315
YEp353
pEG202
pGAPZ B
pRS316
YEp354
pESC-HIS
pGAPZ C
pRS403
YEp355
pESC-LEU
pGAPZ(alpha) A
pRS404
YEp356
pESC-TRP
pGAPZ(alpha) B
pRS405
YEp356R
pESC-URA
pGAPZ(alpha) C
pRS406
YEp357
pFA6-kanMX4
pGBD-C1
pRS413
YEp357R
pFA6a
pGBD-C2
pRS414
YEp358
pFA6a-12Pk-kanMX6
pGBD-C3
pRS415
YEp358R
pFA6a-13Myc-His3MX6
pGBDU-C1
pRS416
YEp363
pFA6a-13Myc-kanMX6
pGBDU-C2
pRS423
YEp364
pFA6a-13Myc-TRP1
pGBDU-C3
pRS424
YEp365
pFA6a-3HA-His3MX6
pGBKT7
pRS425
YEp366
pFA6a-3HA-kanMX6
pGilda
pRS426
YEp366R
pFA6a-3HA-TRP1
pHIL-D2
pSpot5
YEp367
pFA6a-5FLAG-kanMX6
pHIL-S1
pSpot6
YEp367R
pFA6a-bleMX6
pJG4-5
pSpot7
YEp368
pFA6a-GFP(S65T)-His3MX6
pKLAC1
pSpot8
YEp368R
pFA6a-GFP(S65T)-kanMX6
pKLAC2
pTEF1 Zeo
YEplac112
pFA6a-GFP(S65T)-TRP1
pOAD
pUCATPH
YEplac181
pFA6a-GST-His3MX6
pOBD2
pY-secSUMOstar
YEplac195
pFA6a-GST-kanMX6
pP-secSUMO3
pY-SUMOstar
YIp351
pFA6a-GST-TRP1
pPIC3
pYC2-E
YIp352
pFA6a-His3MX6
pPIC3.5
pYC2 CT
YIp5
pFA6a-His3MX6-PGAL1
pPIC3.5K
pYC2 NT A
YIplac128
pFA6a-His3MX6-PGAL1-3HA
pPIC6 A
pYC2 NT B
YIplac204
pFA6a-His3MX6-PGAL1-GFP
pPIC6 B
pYC2 NT C
YIplac211
pFA6a-His3MX6-PGAL1-GST
pPIC6 C
pYC6 CT
YRp7
pFA6a-hphMX6
pPIC6(alpha) A
pYES2
alpha-factor secretion signal

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