pP-secSUMO3

Pichia pastoris vector for regulated expression of secreted recombinant proteins with a cleavable 6xHis-SUMO3 tag.

Sequence Author: LifeSensors

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BglII (1) AlwNI (3527) PspFI (3419) BseYI (3415) HaeII (3359) BssSαI (3284) PciI (3111) MluI (3022) BsrGI (3015) EcoRV (2848) DraIII (2763) FseI (2686) SexAI * (2573) SgrAI (2524) SmaI (2500) TspMI - XmaI (2498) AatII (2456) ZraI (2454) BssHII - MauBI (2446) MscI (2413) NcoI (2408) BtgZI (1983) BamHI (1924) AleI (76) BmrI (178) PflMI (190) Eco53kI (206) BanII - SacI (208) DraI - PmeI (413) NsiI (677) BstXI - XcmI (706) MfeI (864) HindIII (872) BstBI (933) BstAPI (995) PsiI (1136) BsrDI (1155) PaeR7I - XhoI (1184) 6xHis EcoNI (1226) PstI - SbfI (1227) BsgI (1328) BsmBI (1494) BsmBI - XbaI (1518) NotI (1525) SacII (1531) Acc65I (1532) KpnI (1536) EcoRI (1538) BfuAI - BspMI (1555) SalI (1561) AccI (1562) 6xHis AgeI (1643) BsiWI (1775) pP-secSUMO3 3840 bp
BglII  (1)
1 site
A G A T C T T C T A G A
AlwNI  (3527)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
PspFI  (3419)
1 site
C C C A G C G G G T C G
BseYI  (3415)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
HaeII  (3359)
1 site
R G C G C Y Y C G C G R
BssSαI  (3284)
1 site
C A C G A G G T G C T C
PciI  (3111)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
MluI  (3022)
1 site
A C G C G T T G C G C A
BsrGI  (3015)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
EcoRV  (2848)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
DraIII  (2763)
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.
FseI  (2686)
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.
SexAI  (2573)
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  (2524)
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  (2500)
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  (2498)
1 site
C C C G G G G G G C C C
XmaI  (2498)
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.
AatII  (2456)
1 site
G A C G T C C T G C A G
ZraI  (2454)
1 site
G A C G T C C T G C A G
BssHII  (2446)
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  (2446)
1 site
C G C G C G C G G C G C G C G C
MscI  (2413)
1 site
T G G C C A A C C G G T
NcoI  (2408)
1 site
C C A T G G G G T A C C
BtgZI  (1983)
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.
BamHI  (1924)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
AleI  (76)
1 site
C A C N N N N G T G G T G N N N N C A C
BmrI  (178)
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.
PflMI  (190)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
Eco53kI  (206)
1 site
G A G C T C C T C G A G
BanII  (208)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (208)
1 site
G A G C T C C T C G A G
DraI  (413)
1 site
T T T A A A A A A T T T
PmeI  (413)
1 site
G T T T A A A C C A A A T T T G
NsiI  (677)
1 site
A T G C A T T A C G T A
BstXI  (706)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
XcmI  (706)
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.
MfeI  (864)
1 site
C A A T T G G T T A A C
HindIII  (872)
1 site
A A G C T T T T C G A A
BstBI  (933)
1 site
T T C G A A A A G C T T
BstAPI  (995)
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.
PsiI  (1136)
1 site
T T A T A A A A T A T T
BsrDI  (1155)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PaeR7I  (1184)
1 site
C T C G A G G A