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Plasmid Files

pHP45Ω-Km

Plasmid carrying an Ω fragment for in vitro insertional mutagenesis (interposon) with a kanamycin resistance gene.

 
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pHP45(omega)-Km.dna
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BbsI (4576) AatII (4520) ZraI (4518) SspI (4402) XmnI (4197) ScaI (4078) PvuI (3968) BglI (3718) BsaI (3659) AhdI (3598) PciI (2705) NdeI (2528) BstZ17I (2478) BsmBI (2348) HindIII (2295) BspDI - ClaI (2290) EcoRI (1) SmaI (8) BamHI (11) HindIII (32) DraIII (446) XcmI (776) BglII (796) BclI * (801) EagI (866) MscI (1042) BssHII (1357) SphI (1365) BtgI - NcoI (1392) RsrII (1476) BstBI (1642) SmaI (1799) NruI (1815) Bsu36I (1836) SalI (1965) HindIII (2233) BamHI (2254) SmaI (2261) EcoRI (2264) pHP45 Ω- Km 4590 bp
BbsI  (4576)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
AatII  (4520)
1 site
G A C G T C C T G C A G
ZraI  (4518)
1 site
G A C G T C C T G C A G
SspI  (4402)
1 site
A A T A T T T T A T A A
XmnI  (4197)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (4078)
1 site
A G T A C T T C A T G A
PvuI  (3968)
1 site
C G A T C G G C T A G C
BglI  (3718)
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.
BsaI  (3659)
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  (3598)
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.
PciI  (2705)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NdeI  (2528)
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  (2478)
1 site
G T A T A C C A T A T G
BsmBI  (2348)
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.
HindIII  (2295)
3 sites
A A G C T T T T C G A A
BspDI  (2290)
1 site
A T C G A T T A G C T A
ClaI  (2290)
1 site
A T C G A T T A G C T A
EcoRI  (1)
2 sites
G A A T T C C T T A A G
SmaI  (8)
3 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)
2 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.
XcmI  (776)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BglII  (796)
1 site
A G A T C T T C T A G A
BclI  (801)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
EagI  (866)
1 site
C G G C C G G C C G G C
MscI  (1042)
1 site
T G G C C A A C C G G T
BssHII  (1357)
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.
SphI  (1365)
1 site
G C A T G C C G T A C G
BtgI  (1392)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1392)
1 site
C C A T G G G G T A C C
RsrII  (1476)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BstBI  (1642)
1 site
T T C G A A A A G C T T
SmaI  (1799)
3 sites
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
NruI  (1815)
1 site
T C G C G A A G C G C T
Bsu36I  (1836)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
SalI  (1965)
1 site
G T C G A C C A G C T G
HindIII  (2233)
3 sites
A A G C T T T T C G A A
BamHI  (2254)
2 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  (2261)
3 sites
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (2264)
2 sites
G A A T T C C T T A A G
Ω-Km
32 .. 2238  =  2207 bp
insertional mutagenesis element with a kanamycin resistance gene (Fellay et al., 1987)
Ω-Km
32 .. 2238  =  2207 bp
insertional mutagenesis element with a kanamycin resistance gene (Fellay et al., 1987)
AmpR
3525 .. 4385  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3525 .. 4316  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3525 .. 4385  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   4317 .. 4385  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics