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Plasmid Files

pKatushka2S-B

Vector for expressing Katushka2S in bacteria.

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pKatushka2S-B.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Evrogen
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AvaI - BsoBI - PaeR7I - XhoI (1) EcoO109I (4122) AatII (4068) ZraI (4066) XmnI (3745) PvuI (3516) FspI (3368) AseI (3318) AhdI (3146) AflIII - PciI (2253) BspQI - SapI (2137) BstAPI (2077) NdeI (2076) BmeT110I (2) PsiI (49) MfeI (59) BseRI (114) BsaBI * (125) BamHI (126) AgeI (134) AleI (147) BstXI (149) BclI * (169) BsrGI (196) Bsu36I (412) SfiI (622) BsgI (651) PshAI (786) EagI - NotI (856) HindIII (863) NheI (983) BmtI (987) Bpu10I (1008) BspEI (1231) PflMI * (1467) XbaI (1840) PfoI (1897) PflFI - Tth111I (2000) BsaAI (2007) AccI (2025) BstZ17I (2026) pKatushka2S-B 4138 bp
AvaI  (1)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (1)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (1)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1)
1 site
C T C G A G G A G C T C
EcoO109I  (4122)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
AatII  (4068)
1 site
G A C G T C C T G C A G
ZraI  (4066)
1 site
G A C G T C C T G C A G
XmnI  (3745)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (3516)
1 site
C G A T C G G C T A G C
FspI  (3368)
1 site
T G C G C A A C G C G T
AseI  (3318)
1 site
A T T A A T T A A T T A
AhdI  (3146)
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.
AflIII  (2253)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2253)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2137)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (2137)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
BstAPI  (2077)
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.
NdeI  (2076)
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.
BmeT110I  (2)
1 site
C Y C G R G G R G C Y C
PsiI  (49)
1 site
T T A T A A A A T A T T
MfeI  (59)
1 site
C A A T T G G T T A A C
BseRI  (114)
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.
BsaBI  (125)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BamHI  (126)
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.
AgeI  (134)
1 site
A C C G G T T G G C C A
AleI  (147)
1 site
C A C N N N N G T G G T G N N N N C A C
BstXI  (149)
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.
BclI  (169)
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.
BsrGI  (196)
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.
Bsu36I  (412)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
SfiI  (622)
1 site
G G C C N