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Plasmid Files

hKO

Humanized Kusabira-Orange fluorescent protein.

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hKO Sequence and MaphKO.dna
Map and Sequence File   
Sequence Author:  MBL International
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 600 400 200 End (657) Bts α I (648) BsaHI (638) EcoP15I (623) TaqII (622) BbsI (610) PflMI (594) NaeI (501) NgoMIV (499) BmgBI (477) AarI - BfuAI - BspMI (460) BmrI (432) BanII (431) AlwNI (422) BsiHKAI (356) BglI (338) HaeII (323) BpmI (271) Bpu10I (268) StuI (253) SexAI * (173) StyI (143) MscI (142) BaeGI - Bme1580I (91) BanI (88) BssS α I (64) ScaI - XmnI (37) TatI (35) BclI * (11) Start (0) hKO 657 bp
End  (657)
0 sites
BtsαI  (648)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
BsaHI  (638)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
EcoP15I  (623)
1 site
C A G C A G ( N ) 25 G T C G T C ( N ) 25 N N

Efficient cleavage requires two inversely oriented copies of the
EcoP15I recognition sequence.
Sticky ends from different EcoP15I sites may not be compatible.
EcoP15I requires ATP for activity.
TaqII  (622)
1 site
G A C C G A ( N ) 9 N N C T G G C T ( N ) 9

Sticky ends from different TaqII sites may not be compatible.
BbsI  (610)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
PflMI  (594)
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.
NaeI  (501)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
NgoMIV  (499)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
BmgBI  (477)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
AarI  (460)
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 AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BfuAI  (460)
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  (460)
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.
BmrI  (432)
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.
BanII  (431)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
AlwNI  (422)
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.
BsiHKAI  (356)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI sites may not be compatible.
BglI  (338)
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.
HaeII  (323)
1 site
R G C G C Y Y C G C G R
BpmI  (271)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
Bpu10I  (268)
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.
StuI  (253)
1 site
A G G C C T T C C G G A
SexAI  (173)
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.
StyI  (143)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
MscI  (142)
1 site
T G G C C A A C C G G T
BaeGI  (91)
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  (91)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
BanI  (88)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
BssSαI  (64)
1 site
C A C G A G G T G C T C
ScaI  (37)
1 site
A G T A C T T C A T G A
XmnI  (37)
1 site
G A A N N N N T T C C T T N N N N A A G
TatI  (35)
1 site
W G T A C W W C A T G W
BclI  (11)
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.
Start  (0)
0 sites
hKO
1 .. 657  =  657 bp
218 amino acids  =  24.4 kDa
Product: humanized Kusabira-Orange fluorescent
protein
mammalian codon-optimized
hKO
1 .. 657  =  657 bp
218 amino acids  =  24.4 kDa
Product: humanized Kusabira-Orange fluorescent
protein
mammalian codon-optimized
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