pOSIP-KC

Prokaryotic one-step cloning and chromosomal integration vector encoding a kanamycin resistance marker and the phage φC31 integrase.
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Eco53kI (9) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator BsgI (6330) PflFI - Tth111I (6316) PfoI (6213) BtgZI (6093) λ tL3 terminator BmtI (5844) NheI (5840) BglI (5816) NdeI (5609) FRT DraI (5434) SnaBI (5196) PvuI (4655) SspI (4580) EcoNI (4567) NruI (4312) DraIII (4124) AfeI (3764) BspEI (3746) SacI (11) Acc65I (13) KpnI (17) BamHI (22) MCS 1 BstZ17I (111) BsrGI (198) SrfI (225) MslI (340) BstXI (342) BsaI (363) BglII (469) SpeI (1126) SphI (1150) PaeR7I - XhoI (1152) MfeI (1239) PacI (1340) AgeI (1343) PspOMI (1349) ApaI (1353) ZraI (1379) AatII (1381) AvrII (1418) BstAPI (1965) PshAI (2090) SbfI (2245) NmeAIII (2272) PasI (2604) MluI (2690) FspI (2867) RsrII (3250) BspDI * - ClaI * (3409) pOSIP-KC 6970 bp
Eco53kI  (9)
1 site
G A G C T C C T C G A G
BsgI  (6330)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PflFI  (6316)
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  (6316)
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.
PfoI  (6213)
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.
BtgZI  (6093)
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.
BmtI  (5844)
1 site
G C T A G C C G A T C G
NheI  (5840)
1 site
G C T A G C C G A T C G
BglI  (5816)
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.
NdeI  (5609)
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.
DraI  (5434)
1 site
T T T A A A A A A T T T
SnaBI  (5196)
1 site
T A C G T A A T G C A T
PvuI  (4655)
1 site
C G A T C G G C T A G C
SspI  (4580)
1 site
A A T A T T T T A T A A
EcoNI  (4567)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
NruI  (4312)
1 site
T C G C G A A G C G C T
DraIII  (4124)
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.
AfeI  (3764)
1 site
A G C G C T T C G C G A
BspEI  (3746)
1 site
T C C G G A A G G C C T
SacI  (11)
1 site
G A G C T C C T C G A G
Acc65I  (13)
1 site
G G T A C C C C A T G G
KpnI  (17)
1 site
G G T A C C C C A T G G
BamHI  (22)
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.
BstZ17I  (111)
1 site
G T A T A C C A T A T G
BsrGI  (198)
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.
SrfI  (225)
1 site
G C C C G G G C C G G G C C C G
MslI  (340)
1 site
C A Y N N N N R T G G T R N N N N Y A C
BstXI  (342)
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.
BsaI  (363)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BglII  (469)
1 site
A G A T C T T C T A G A
SpeI  (1126)
1 site
A C T A G T T G A T C A
SphI  (1150)
1 site
G C A T G C C G T A C G
PaeR7I  (1152)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1152)
1 site
C T C G A G G A G C T C
MfeI  (1239)
1 site
C A A T T G G T T A A C
PacI  (1340)
1 site
T T A A T T A A A A T T A A T T
AgeI  (1343)
1 site
A C C G G T T G G C C A
PspOMI  (1349)
1 site
G G G C C C C C C G G G
ApaI  (1353)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
ZraI  (1379)
1 site
G A C G T C C T G C A G
AatII  (1381)
1 site
G A C G T C C T G C A G
AvrII  (1418)
1 site
C C T A G G G G A T C C
BstAPI  (1965)
1 site
G C A N N N N N T G C C G T N N N