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Plasmid Files

pOSIP-TT

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

 
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 Acc65I (13) EcoRI (1) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator PflFI - Tth111I (6566) BmtI (6094) NheI (6090) SnaBI (5099) SalI (5014) ScaI (4481) PmlI (4204) BspDI - ClaI (3582) KpnI (17) BamHI (22) BssHII (70) SrfI (225) BglII (469) SpeI (1126) PstI (1144) PaeR7I - XhoI (1152) PacI (1340) PspOMI (1349) ApaI (1353) AvrII (1418) EarI (1442) PshAI (2090) Bts α I (2887) SexAI * (3200) DraIII (3576) pOSIP-TT 7395 bp
Acc65I  (13)
1 site
G G T A C C C C A T G G
EcoRI  (1)
1 site
G A A T T C C T T A A G
PflFI  (6566)
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  (6566)
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.
BmtI  (6094)
1 site
G C T A G C C G A T C G
NheI  (6090)
1 site
G C T A G C C G A T C G
SnaBI  (5099)
1 site
T A C G T A A T G C A T
SalI  (5014)
1 site
G T C G A C C A G C T G
ScaI  (4481)
1 site
A G T A C T T C A T G A
PmlI  (4204)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BspDI  (3582)
1 site
A T C G A T T A G C T A
ClaI  (3582)
1 site
A T C G A T T A G C T A
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
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
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.
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.
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.
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.
TcR
3649 .. 4854  =  1206 bp
401 amino acids  =  43.4 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
3649 .. 4854  =  1206 bp
401 amino acids  =  43.4 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
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
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
5590 .. 6089  =  500 bp
attachment site of phage φ21
phage φ21 attP
5590 .. 6089  =  500 bp
attachment site of phage φ21
R6K γ ori
5064 .. 5452  =  389 bp
γ replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication
R6K γ ori
5064 .. 5452  =  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
6110 .. 6356  =  247 bp
transcription terminator tL3 from phage λ
λ tL3 terminator
6110 .. 6356  =  247 bp
transcription terminator tL3 from phage λ
lambda t0 terminator
4913 .. 5007  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
4913 .. 5007  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
6676 .. 6762  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6676 .. 6762  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6857 .. 6943  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
6857 .. 6943  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
7038 .. 7124  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
7038 .. 7124  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
7219 .. 7305  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
7219 .. 7305  =  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
3587 .. 3641  =  55 bp
overlapping promoters for bacterial tetR and tetA
tetR/tetA promoters
3587 .. 3641  =  55 bp
overlapping promoters for bacterial tetR and tetA
FRT
5463 .. 5510  =  48 bp
FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011).
FRT
5463 .. 5510  =  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
3622 .. 3640  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
3622 .. 3640  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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