pHP45(omega)-Tc

Plasmid carrying an Ω fragment for in vitro insertional mutagenesis (interposon) with a tetracycline resistance gene.
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AatII (4427) ZraI (4425) SspI (4309) XmnI (4104) ScaI (3985) PvuI (3875) PstI (3750) BsaI (3566) AhdI (3505) AlwNI (3028) AflIII - PciI (2612) BspQI - SapI (2496) NdeI (2435) BstZ17I (2385) PflFI - Tth111I (2359) PfoI (2256) BsmBI (2255) EcoRI (1) SmaI (8) BamHI (11) HindIII (32) DraIII (446) EcoRV (634) NheI (676) BmtI (680) FspAI (709) BamHI (822) SgrAI (857) BtgI (975) SphI (1013) EcoNI (1073) SalI (1098) PshAI (1163) EagI (1386) NruI (1421) BfuAI - BspMI (1501) BsmI (1806) BsaBI * (1920) HindIII (2140) BamHI (2161) SmaI (2168) EcoRI (2171) BspDI - ClaI (2197) HindIII (2202) pHP45 Ω- Tc 4497 bp
AatII  (4427)
1 site
G A C G T C C T G C A G
ZraI  (4425)
1 site
G A C G T C C T G C A G
SspI  (4309)
1 site
A A T A T T T T A T A A
XmnI  (4104)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3985)
1 site
A G T A C T T C A T G A
PvuI  (3875)
1 site
C G A T C G G C T A G C
PstI  (3750)
1 site
C T G C A G G A C G T C
BsaI  (3566)
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.
AhdI  (3505)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
AlwNI  (3028)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AflIII  (2612)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2612)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2496)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (2496)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
NdeI  (2435)
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.
BstZ17I  (2385)
1 site
G T A T A C C A T A T G
PflFI  (2359)
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  (2359)
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  (2256)
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.
BsmBI  (2255)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
EcoRI  (1)
2 sites
G A A T T C C T T A A G
SmaI  (8)
2 sites
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (11)
3 sites
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.
HindIII  (32)
3 sites
A A G C T T T T C G A A
DraIII  (446)
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.
EcoRV  (634)
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.
NheI  (676)
1 site
G C T A G C C G A T C G
BmtI  (680)
1 site
G C T A G C C G A T C G
FspAI  (709)
1 site
R T G C G C A Y Y A C G C G T R
BamHI  (822)
3 sites
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.
SgrAI  (857)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
BtgI  (975)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SphI  (1013)
1 site
G C A T G C C G T A C G
EcoNI  (1073)
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.
SalI  (1098)
1 site
G T C G A C C A G C T G
PshAI  (1163)
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.
EagI  (1386)
1 site
C G G C C G G C C G G C
NruI  (1421)
1 site
T C G C G A A G C G C T
BfuAI  (1501)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1501)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BsmI  (1806)
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.
BsaBI  (1920)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
HindIII  (2140)
3 sites
A A G C T T T T C G A A
BamHI  (2161)
3 sites
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.
SmaI  (2168)
2 sites
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (2171)
2 sites
G A A T T C C T T A A G
BspDI  (2197)
1 site
A T C G A T T A G C T A
ClaI  (2197)
1 site
A T C G A T T A G C T A
HindIII  (2202)
3 sites
A A G C T T T T C G A A
Ω-Tc
32 .. 2145  =  2114 bp
insertional mutagenesis element with a tetracycline resistance gene (Fellay et al., 1987)
Ω-Tc
32 .. 2145  =  2114 bp
insertional mutagenesis element with a tetracycline resistance gene (Fellay et al., 1987)
AmpR
3432 .. 4292  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   3432 .. 4223  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3432 .. 4292  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   4224 .. 4292  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3432 .. 4292  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2673 .. 3261  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2673 .. 3261  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
bom
2347 .. 2487  =  141 bp
basis of mobility region from pBR322
bom
2347 .. 2487  =  141 bp
basis of mobility region from pBR322
AmpR promoter
4293 .. 4397  =  105 bp
AmpR promoter
4293 .. 4397  =  105 bp
stop codons
19 .. 29  =  11 bp
stop codons in all three reading frames
stop codons
19 .. 29  =  11 bp
stop codons in all three reading frames
stop codons
2148 .. 2158  =  11 bp
stop codons in all three reading frames
stop codons
2148 .. 2158  =  11 bp
stop codons in all three reading frames
TcR
533 .. 1723  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
533 .. 1723  =  1191 bp
396 amino acids  =  41.5 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
T4 transcription termination signals
32 .. 148  =  117 bp
T4 transcription termination signals
32 .. 148  =  117 bp
T4 transcription termination signals
2029 .. 2145  =  117 bp
T4 transcription termination signals
2029 .. 2145  =  117 bp
ORF:  706 .. 1089  =  384 bp
ORF:  127 amino acids  =  14.7 kDa
ORF:  1627 .. 1887  =  261 bp
ORF:  86 amino acids  =  9.7 kDa
ORF:  3562 .. 3828  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  533 .. 1723  =  1191 bp
ORF:  396 amino acids  =  41.5 kDa
ORF:  886 .. 1236  =  351 bp
ORF:  116 amino acids  =  12.2 kDa
ORF:  3432 .. 4292  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  770 .. 1012  =  243 bp
ORF:  80 amino acids  =  8.0 kDa
ORF:  1070 .. 1528  =  459 bp
ORF:  152 amino acids  =  16.7 kDa
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