pM-SUMOstar

Mammalian cell vector for expression of intracellular recombinant proteins with a cleavable 6xHis-SUMOstar tag.

Sequence Author: LifeSensors

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SgrDI (5802) SspI (5685) ScaI (5361) PvuI (5251) AhdI (4881) PciI (3988) BstZ17I (3609) BsmI (3557) PfoI (3412) RsrII (3151) BssHII (3032) MfeI (161) Bpu10I (180) SpeI (249) CMV enhancer NdeI (484) SnaBI (590) Eco53kI (816) SacI (818) HindIII (902) ATG KflI - PpuMI (936) SbfI (945) BlpI (969) BsmBI (1230) BsmBI (1254) NotI (1261) Acc65I (1268) KpnI (1272) PaeR7I - PspXI - XhoI (1273) PspOMI (1285) ApaI (1289) BstBI (1296) V5 tag AgeI (1346) PmeI (1376) BbsI (1587) DraIII (1901) SexAI * (2192) StuI (2424) AvrII (2425) TspMI - XmaI (2446) SmaI (2448) BsaBI * (2494) KasI (2634) NarI (2635) SfoI (2636) PluTI (2638) MscI (2717) PflFI - Tth111I (2753) pM-SUMOstar 5804 bp
SgrDI  (5802)
1 site
C G T C G A C G G C A G C T G C
SspI  (5685)
1 site
A A T A T T T T A T A A
ScaI  (5361)
1 site
A G T A C T T C A T G A
PvuI  (5251)
1 site
C G A T C G G C T A G C
AhdI  (4881)
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.
PciI  (3988)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (3609)
1 site
G T A T A C C A T A T G
BsmI  (3557)
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.
PfoI  (3412)
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.
RsrII  (3151)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BssHII  (3032)
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.
MfeI  (161)
1 site
C A A T T G G T T A A C
Bpu10I  (180)
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.
SpeI  (249)
1 site
A C T A G T T G A T C A
NdeI  (484)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
SnaBI  (590)
1 site
T A C G T A A T G C A T
Eco53kI  (816)
1 site
G A G C T C C T C G A G
SacI  (818)
1 site
G A G C T C C T C G A G
HindIII  (902)
1 site
A A G C T T T T C G A A
KflI  (936)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (936)
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.
SbfI  (945)
1 site
C C T G C A G G G G A C G T C C
BlpI  (969)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BsmBI  (1230)
2 sites
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.
BsmBI  (1254)
2 sites
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.
NotI  (1261)
1 site
G C G G C C G C C G C C G G C G
Acc65I  (1268)
1 site
G G T A C C C C A T G G
KpnI  (1272)
1 site
G G T A C C C C A T G G
PaeR7I  (1273)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1273)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1273)
1 site
C T C G A G G A G C T C
PspOMI  (1285)
1 site
G G G C C C C C C G G G
ApaI  (1289)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BstBI  (1296)
1 site
T T C G A A A A G C T T
AgeI  (1346)
1 site
A C C G G T T G G C C A
PmeI  (1376)
1 site
G T T