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Plasmid Files

pOSIP-TH

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

 
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 EcoRI (1) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator PflFI - Tth111I (6096) BseRI (5748) BmtI (5624) NheI (5620) NdeI (5351) SalI (4853) StuI (4714) ScaI (4320) BglI (4277) PmlI (4043) SspI (3991) EcoRV (3601) BspDI - ClaI (3421) Acc65I (13) KpnI (17) BamHI (22) BssHII (70) SrfI (225) BsaI (363) BglII (469) BciVI (923) SpeI (1126) PstI (1144) SphI (1150) PaeR7I - XhoI (1152) BstBI (1234) PacI (1340) PspOMI (1349) ApaI (1353) ZraI (1379) AatII (1381) AvrII (1418) BsmI (1718) PshAI (2090) KflI - PpuMI (2350) NruI (2828) Bts α I (3076) DraIII (3415) pOSIP-TH 6925 bp
EcoRI  (1)
1 site
G A A T T C C T T A A G
PflFI  (6096)
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  (6096)
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  (5748)
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  (5624)
1 site
G C T A G C C G A T C G
NheI  (5620)
1 site
G C T A G C C G A T C G
NdeI  (5351)
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  (4853)
1 site
G T C G A C C A G C T G
StuI  (4714)
1 site
A G G C C T T C C G G A
ScaI  (4320)
1 site
A G T A C T T C A T G A
BglI  (4277)
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.
PmlI  (4043)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
SspI  (3991)
1 site
A A T A T T T T A T A A
EcoRV  (3601)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
BspDI  (3421)
1 site
A T C G A T T A G C T A
ClaI  (3421)
1 site
A T C G A T T A G C T A
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.
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.
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
BstBI  (1234)
1 site
T T C G A A A A G C T T
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.
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
BsmI  (1718)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI 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.
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.
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.
TcR
3488 .. 4693  =  1206 bp
401 amino acids  =  43.4 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
3488 .. 4693  =  1206 bp
401 amino acids  =  43.4 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
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
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
4903 .. 5291  =  389 bp
γ replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication
R6K γ ori
4903 .. 5291  =  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
5640 .. 5886  =  247 bp
transcription terminator tL3 from phage λ
λ tL3 terminator
5640 .. 5886  =  247 bp
transcription terminator tL3 from phage λ
HK022 attP
5379 .. 5605  =  227 bp
attachment site of phage HK022
HK022 attP
5379 .. 5605  =  227 bp
attachment site of phage HK022
lambda t0 terminator
4752 .. 4846  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
4752 .. 4846  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
6206 .. 6292  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6206 .. 6292  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6387 .. 6473  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6387 .. 6473  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6568 .. 6654  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6568 .. 6654  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6749 .. 6835  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6749 .. 6835  =  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
tetR/tetA promoters
3426 .. 3480  =  55 bp
overlapping promoters for bacterial tetR and tetA
tetR/tetA promoters
3426 .. 3480  =  55 bp
overlapping promoters for bacterial tetR and tetA
FRT
5302 .. 5349  =  48 bp
FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011).
FRT
5302 .. 5349  =  48 bp
FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011).
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)
MCS 2
1126 .. 1157  =  32 bp
multiple cloning site, part 2
MCS 2
1126 .. 1157  =  32 bp
multiple cloning site, part 2
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
tet operator
3461 .. 3479  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
3461 .. 3479  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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