Resources
Plasmid Files

pOSIP-KC

Prokaryotic one-step cloning and chromosomal integration vector encoding a kanamycin resistance marker and the phage φC31 integrase.

 
To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

 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

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
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 N N A C G

Sticky ends from different BstAPI sites may not be compatible.
PshAI  (2090)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SbfI  (2245)
1 site
C C T G C A G G G G A C G T C C
NmeAIII  (2272)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PasI  (2604)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
MluI  (2690)
1 site
A C G C G T T G C G C A
FspI  (2867)
1 site
T G C G C A A C G C G T
RsrII  (3250)
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.
BspDI  (3409)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3409)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
phage φC31 integrase
2272 .. 4089  =  1818 bp
605 amino acids  =  67.1 kDa
Product: integrase from phage φC31
phage φC31 integrase
2272 .. 4089  =  1818 bp
605 amino acids  =  67.1 kDa
Product: integrase from phage φC31
KanR
4224 .. 5039  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
4224 .. 5039  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
λ repressor (ts)
1424 .. 2137  =  714 bp
237 amino acids  =  26.2 kDa
Product: temperature-sensitive variant of the phage
λ repressor
thermosensitivity is conferred by the A67T mutation
λ repressor (ts)
1424 .. 2137  =  714 bp
237 amino acids  =  26.2 kDa
Product: temperature-sensitive variant of the phage
λ repressor
thermosensitivity is conferred by the A67T mutation
ori
477 .. 1065  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
477 .. 1065  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
R6K γ ori
5161 .. 5549  =  389 bp
γ replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication
R6K γ ori
5161 .. 5549  =  389 bp
γ replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication
ccdB
142 .. 447  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
ccdB
142 .. 447  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
λ tL3 terminator
5860 .. 6106  =  247 bp
transcription terminator tL3 from phage λ
λ tL3 terminator
5860 .. 6106  =  247 bp
transcription terminator tL3 from phage λ
phage φC31 attP
5676 .. 5775  =  100 bp
attachment site of phage φC31
phage φC31 attP
5676 .. 5775  =  100 bp
attachment site of phage φC31
rrnB T1 terminator
6428 .. 6514  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6428 .. 6514  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6611 .. 6697  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6611 .. 6697  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6794 .. 6880  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6794 .. 6880  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
MCS 1
1 .. 75  =  75 bp
multiple cloning site, part 1
MCS 1
1 .. 75  =  75 bp
multiple cloning site, part 1
FRT
5560 .. 5607  =  48 bp
FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011).
FRT
5560 .. 5607  =  48 bp
FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011).
MCS 2
1126 .. 1169  =  44 bp
multiple cloning site, part 2
MCS 2
1126 .. 1169  =  44 bp
multiple cloning site, part 2
FRT (minimal)
1384 .. 1417  =  34 bp
supports FLP-mediated excision but not integration
(Turan and Bode, 2011)
FRT (minimal)
1384 .. 1417  =  34 bp
supports FLP-mediated excision but not integration
(Turan and Bode, 2011)
tonB terminator
1276 .. 1307  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
tonB terminator
1276 .. 1307  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
rrnB T2 terminator
1200 .. 1227  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
1200 .. 1227  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter