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Plasmid Files

pOSIP-CH

Prokaryotic one-step cloning and chromosomal integration vector encoding a chloramphenicol resistance marker and the HK022 integrase.

 
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 Acc65I (13) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator PflFI - Tth111I (5569) BseRI (5221) BmtI (5097) NheI (5093) NdeI (4824) SalI (4326) ScaI (4175) SspI (4070) PasI (3990) BpmI (3880) BspEI (3754) PvuII (3658) DraIII (3415) KpnI (17) BamHI (22) BssHII (70) BsrGI (198) SrfI (225) BsaI (363) BglII (469) BciVI (923) SpeI (1126) PstI (1144) SphI (1150) PaeR7I - XhoI (1152) KasI (1164) NarI (1165) SfoI (1166) PluTI (1168) BstBI (1234) MfeI (1239) PacI (1340) AgeI (1343) PspOMI (1349) ApaI (1353) ZraI (1379) AatII (1381) AvrII (1418) BstAPI (1965) PshAI (2090) AflIII - MluI (2240) KflI - PpuMI (2350) NruI (2828) Bts α I (3076) pOSIP-CH 6398 bp
Acc65I  (13)
1 site
G G T A C C C C A T G G
PflFI  (5569)
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  (5569)
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.
BseRI  (5221)
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.
BmtI  (5097)
1 site
G C T A G C C G A T C G
NheI  (5093)
1 site
G C T A G C C G A T C G
NdeI  (4824)
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.
SalI  (4326)
1 site
G T C G A C C A G C T G
ScaI  (4175)
1 site
A G T A C T T C A T G A
SspI  (4070)
1 site
A A T A T T T T A T A A
PasI  (3990)
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  (3880)
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  (3754)
1 site
T C C G G A A G G C C T
PvuII  (3658)
1 site
C A G C T G G T C G A C
DraIII  (3415)
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
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
BciVI  (923)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
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
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
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.
BstBI  (1234)
1 site
T T C G A A A A G C T T
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.
AflIII  (2240)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (2240)
1 site
A C G C G T T G C G C A
KflI  (2350)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (2350)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
NruI  (2828)
1 site
T C G C G A A G C G C T
BtsαI  (3076)
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.
HK022 integrase
2270 .. 3343  =  1074 bp
357 amino acids  =  40.4 kDa
Product: integrase from phage HK022
HK022 integrase
2270 .. 3343  =  1074 bp
357 amino acids  =  40.4 kDa
Product: integrase from phage HK022
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
3545 .. 4204  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3545 .. 4204  =  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
R6K γ ori
4376 .. 4764  =  389 bp
γ replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication
R6K γ ori
4376 .. 4764  =  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
5113 .. 5359  =  247 bp
transcription terminator tL3 from phage λ
λ tL3 terminator
5113 .. 5359  =  247 bp
transcription terminator tL3 from phage λ
HK022 attP
4852 .. 5078  =  227 bp
attachment site of phage HK022
HK022 attP
4852 .. 5078  =  227 bp
attachment site of phage HK022
cat promoter
3442 .. 3544  =  103 bp
promoter of the E. coli cat gene
cat promoter
3442 .. 3544  =  103 bp
promoter of the E. coli cat gene
lambda t0 terminator
4225 .. 4319  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
4225 .. 4319  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
5679 .. 5765  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
5679 .. 5765  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
5860 .. 5946  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
5860 .. 5946  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6041 .. 6127  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6041 .. 6127  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6222 .. 6308  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6222 .. 6308  =  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
4775 .. 4822  =  48 bp
FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011).
FRT
4775 .. 4822  =  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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