pHP45(omega)-Km

Plasmid carrying an Ω fragment for in vitro insertional mutagenesis (interposon) with a kanamycin resistance gene.
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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) 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) BspDI - ClaI (2290) 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.
BsmBI-v2 is an improved version of BsmBI.
HindIII  (2295)
3 sites
A A G C T T T T C G A 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
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
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
2 segments
   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
2 segments
   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
AmpR
3525 .. 4385  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2766 .. 3354  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2766 .. 3354  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
bom
2440 .. 2580  =  141 bp
basis of mobility region from pBR322
bom
2440 .. 2580  =  141 bp
basis of mobility region from pBR322
AmpR promoter
4386 .. 4490  =  105 bp
AmpR promoter
4386 .. 4490  =  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
2241 .. 2251  =  11 bp
stop codons in all three reading frames
stop codons
2241 .. 2251  =  11 bp
stop codons in all three reading frames
NeoR/KanR
832 .. 1626  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
832 .. 1626  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
T4 transcription termination signals
32 .. 148  =  117 bp
T4 transcription termination signals
32 .. 148  =  117 bp
T4 transcription termination signals
2122 .. 2238  =  117 bp
T4 transcription termination signals
2122 .. 2238  =  117 bp
ORF:  832 .. 1626  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  3655 .. 3921  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1004 .. 1390  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  1647 .. 2069  =  423 bp
ORF:  140 amino acids  =  15.5 kDa
ORF:  1141 .. 1677  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  1677 .. 1922  =  246 bp
ORF:  81 amino acids  =  8.4 kDa
ORF:  3525 .. 4385  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
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