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Plasmid Files

pOSIP-CT

Prokaryotic one-step cloning and chromosomal integration vector encoding a chloramphenicol resistance marker and the phage φ21 integrase.

 
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 Acc65I (13) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator PflFI - Tth111I (6039) BbvCI (5703) BmtI (5567) NheI (5563) StuI (5556) BglI (5329) SnaBI (4572) SalI (4487) ScaI (4336) PasI (4151) BpmI (4041) BspEI (3915) PvuII (3819) DraIII (3576) KpnI (17) BamHI (22) BssHII (70) BsrGI (198) SrfI (225) BglII (469) SpeI (1126) PstI (1144) PaeR7I - XhoI (1152) KasI (1164) NarI (1165) SfoI (1166) PluTI (1168) MfeI (1239) PacI (1340) PspOMI (1349) ApaI (1353) AvrII (1418) EarI (1442) BstAPI (1965) PshAI (2090) MluI (2240) Bts α I (2887) SexAI * (3200) pOSIP-CT 6868 bp
Acc65I  (13)
1 site
G G T A C C C C A T G G
PflFI  (6039)
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  (6039)
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.
BbvCI  (5703)
1 site
C C T C A G C G G A G T C G
BmtI  (5567)
1 site
G C T A G C C G A T C G
NheI  (5563)
1 site
G C T A G C C G A T C G
StuI  (5556)
1 site
A G G C C T T C C G G A
BglI  (5329)
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.
SnaBI  (4572)
1 site
T A C G T A A T G C A T
SalI  (4487)
1 site
G T C G A C C A G C T G
ScaI  (4336)
1 site
A G T A C T T C A T G A
PasI  (4151)
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.
BpmI  (4041)
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.
BspEI  (3915)
1 site
T C C G G A A G G C C T
PvuII  (3819)
1 site
C A G C T G G T C G A C
DraIII  (3576)
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.
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.
BssHII  (70)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
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
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
PstI  (1144)
1 site
C T G C A G G A C G T C
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
KasI  (1164)
1 site
G G C G C C C C G C G G
NarI  (1165)
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.
SfoI  (1166)
1 site
G G C G C C C C G C G G
PluTI  (1168)
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.
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
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.
AvrII  (1418)
1 site
C C T A G G G G A T C C
EarI  (1442)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI
recognition sequence.
Sticky ends from different EarI sites may not be compatible.
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.
MluI  (2240)
1 site
A C G C G T T G C G C A
BtsαI  (2887)
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.
SexAI  (3200)
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.
phage φ21 integrase
2299 .. 3441  =  1143 bp
380 amino acids  =  43.9 kDa
Product: integrase from phage φ21
phage φ21 integrase
2299 .. 3441  =  1143 bp
380 amino acids  =  43.9 kDa
Product: integrase from phage φ21
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
CmR
3706 .. 4365  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3706 .. 4365  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
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
phage φ21 attP
5063 .. 5562  =  500 bp
attachment site of phage φ21
phage φ21 attP
5063 .. 5562  =  500 bp
attachment site of phage φ21
R6K γ ori
4537 .. 4925  =  389 bp
γ replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication
R6K γ ori
4537 .. 4925  =  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
5583 .. 5829  =  247 bp
transcription terminator tL3 from phage λ
λ tL3 terminator
5583 .. 5829  =  247 bp
transcription terminator tL3 from phage λ
cat promoter
3603 .. 3705  =  103 bp
promoter of the E. coli cat gene
cat promoter
3603 .. 3705  =  103 bp
promoter of the E. coli cat gene
lambda t0 terminator
4386 .. 4480  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
4386 .. 4480  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
6149 .. 6235  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6149 .. 6235  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6330 .. 6416  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6330 .. 6416  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6511 .. 6597  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6511 .. 6597  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6692 .. 6778  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6692 .. 6778  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
MCS 1
13 .. 75  =  63 bp
multiple cloning site, part 1
MCS 1
13 .. 75  =  63 bp
multiple cloning site, part 1
FRT
4936 .. 4983  =  48 bp
FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011).
FRT
4936 .. 4983  =  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
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