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Plasmid Files

lacZ

E. coli lacZ gene encoding β-galactosidase.

 
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 3000 2000 1000 End (3075) BlpI (3028) EcoRI (3019) PfoI * (2995) NdeI (2972) BsiWI (2786) BstZ17I (2780) AccI (2779) NspI (2778) PciI (2774) BstYI (2757) BsaWI (2570) DrdI (2124) SacI (1955) Eco53kI (1953) AfeI (1850) BciVI (1688) BssHII - MauBI (1513) BclI * (1361) BsaBI * (1346) SspI (1245) DraIII (1205) EcoRV (1128) BssS α I (1072) BspDI - ClaI (839) AcuI (749) TatI (717) AatII (638) ZraI (636) Bpu10I (520) NmeAIII (425) Bsu36I (239) FspI (157) EarI (116) Start (0) lacZ 3075 bp
End  (3075)
0 sites
BlpI  (3028)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
EcoRI  (3019)
1 site
G A A T T C C T T A A G
PfoI  (2995)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
NdeI  (2972)
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.
BsiWI  (2786)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BstZ17I  (2780)
1 site
G T A T A C C A T A T G
AccI  (2779)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
NspI  (2778)
1 site
R C A T G Y Y G T A C R
PciI  (2774)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstYI  (2757)
1 site
R G A T C Y Y C T A G R
BsaWI  (2570)
1 site
W C C G G W W G G C C W

Efficient cleavage requires at least two copies of the BsaWI
recognition sequence.
DrdI  (2124)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
SacI  (1955)
1 site
G A G C T C C T C G A G
Eco53kI  (1953)
1 site
G A G C T C C T C G A G
AfeI  (1850)
1 site
A G C G C T T C G C G A
BciVI  (1688)
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.
BssHII  (1513)
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.
MauBI  (1513)
1 site
C G C G C G C G G C G C G C G C
BclI  (1361)
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.
BsaBI  (1346)
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.
SspI  (1245)
1 site
A A T A T T T T A T A A
DraIII  (1205)
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  (1128)
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.
BssSαI  (1072)
1 site
C A C G A G G T G C T C
BspDI  (839)
1 site
A T C G A T T A G C T A
ClaI  (839)
1 site
A T C G A T T A G C T A
AcuI  (749)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI
recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
TatI  (717)
1 site
W G T A C W W C A T G W
AatII  (638)
1 site
G A C G T C C T G C A G
ZraI  (636)
1 site
G A C G T C C T G C A G
Bpu10I  (520)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
NmeAIII  (425)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
Bsu36I  (239)
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.
FspI  (157)
1 site
T G C G C A A C G C G T
EarI  (116)
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.
Start  (0)
0 sites
lacZ
1 .. 3075  =  3075 bp
1024 amino acids  =  116.5 kDa
Product: β-galactosidase
lacZ
1 .. 3075  =  3075 bp
1024 amino acids  =  116.5 kDa
Product: β-galactosidase
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