pKillerOrange-C

Vector for fusing KillerOrange to the N-terminus of a partner protein.

Sequence Author: Evrogen

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PfoI (3524) RsrII (3265) BsrDI (2982) PflFI - Tth111I (2867) FspI (2851) MscI (2831) PluTI (2752) SfoI (2750) NarI (2749) KasI (2748) EagI (2655) BspDI * - ClaI * (2589) StuI (2570) BseRI (2567) SfiI (2524) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (600) Bpu10I (880) BlpI (946) PaqCI (964) PmlI (1251) BspEI (1324) BglII (1333) PaeR7I - XhoI (1337) Eco53kI (1342) SacI (1344) HindIII (1346) EcoRI (1353) PstI (1362) SalI (1363) AccI (1364) Acc65I (1369) KpnI (1373) SacII (1376) PspOMI (1377) TspMI - XmaI (1380) ApaI (1381) SmaI (1382) BamHI (1384) XbaI * (1396) BclI * (1406) MfeI (1499) HpaI (1512) BtsI - BtsαI (1588) MluI (1635) CsiI - SexAI * (2338) pKillerOrange-C 4725 bp
PfoI  (3524)
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  (3265)
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.
BsrDI  (2982)
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.
PflFI  (2867)
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  (2867)
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.
FspI  (2851)
1 site
T G C G C A A C G C G T
MscI  (2831)
1 site
T G G C C A A C C G G T
PluTI  (2752)
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  (2750)
1 site
G G C G C C C C G C G G
NarI  (2749)
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  (2748)
1 site
G G C G C C C C G C G G
EagI  (2655)
1 site
C G G C C G G C C G G C
BspDI  (2589)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2589)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2570)
1 site
A G G C C T T C C G G A
BseRI  (2567)
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.
SfiI  (2524)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
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  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
AfeI  (596)
1 site
A G C G C T T C G C G A
AgeI  (600)
1 site
A C C G G T T G G C C A
Bpu10I  (880)
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.
BlpI  (946)
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.
PaqCI  (964)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
PmlI  (1251)
1 site
C A C G T G G T G C A C
BspEI  (1324)
1 site
T C C G G A A G G C C T
BglII  (1333)
1 site
A G A T C T T C T A G A
PaeR7I  (1337)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1337)
1 site
C T C G A G G A G C T C
Eco53kI  (1342)
1 site
G A G C T C C T C G A G
SacI  (1344)
1 site
G A G C T C C T C G A G
HindIII  (1346)
1 site
A A G C T T T T C G A A
EcoRI  (1353)
1 site
G A A T T C C T T A A G
PstI  (1362)
1 site
C T G C A G G A C G T C
SalI  (1363)
1 site
G T C G A C C A G C T G
AccI  (1364)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
Acc65I  (1369)
1 site
G G T A C C C C A T G G
KpnI  (1373)
1 site
G G T A C C C C A T G G
SacII  (1376)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
PspOMI  (1377)
1 site
G G G C C C C C C G G G
TspMI  (1380)
1 site
C C C G G G G G G C C C
XmaI  (1380)
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.
ApaI  (1381)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SmaI  (1382)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (1384)
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.
XbaI  (1396)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1406)
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.
MfeI  (1499)
1 site
C A A T T G G T T A A C
HpaI  (1512)
1 site
G T T A A C C A A T T G
BtsI  (1588)
1 site
G C A G T G N N C G T C A C
BtsαI  (1588)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
MluI  (1635)
1 site
A C G C G T T G C G C A
CsiI  (2338)
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  (2338)
1 site
A C C W G G T T G G W C