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Plasmid Files

pLacI

lac repressor plasmid with a p15A origin of replication.

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pLacI Sequence and MappLacI.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
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 MscI (3551) NcoI - StyI (3513) ScaI (3401) TatI (3399) DrdI - PflFI - Tth111I (3270) Bsu36I (3161) BsaBI * (3014) EcoO109I (2957) NaeI (2928) NgoMIV (2926) EcoNI (2812) BstAPI (2668) ApaLI (2364) AflIII - MluI (2344) BstEII (2162) NmeAIII (2144) ApaI - BanII (2141) PspOMI (2137) BssHII (1933) EcoRI (1) BsaAI (312) NheI (583) BmtI (587) AccI (595) BstZ17I (596) XmnI (639) SgrAI (669) BsrBI (757) SacII (835) BssS α I (959) NspI (1161) XbaI (1424) AcuI (1484) KasI (1705) NarI * (1706) SfoI (1707) PluTI (1709) HincII - HpaI (1842) BciVI (1892) pLacI 3813 bp
MscI  (3551)
1 site
T G G C C A A C C G G T
NcoI  (3513)
1 site
C C A T G G G G T A C C
StyI  (3513)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
ScaI  (3401)
1 site
A G T A C T T C A T G A
TatI  (3399)
1 site
W G T A C W W C A T G W
DrdI  (3270)
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.
PflFI  (3270)
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  (3270)
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.
Bsu36I  (3161)
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.
BsaBI  (3014)
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.
EcoO109I  (2957)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
NaeI  (2928)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
NgoMIV  (2926)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
EcoNI  (2812)
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.
BstAPI  (2668)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
ApaLI  (2364)
1 site
G T G C A C C A C G T G
AflIII  (2344)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (2344)
1 site
A C G C G T T G C G C A
BstEII  (2162)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
NmeAIII  (2144)
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).
ApaI  (2141)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (2141)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PspOMI  (2137)
1 site
G G G C C C C C C G G G
BssHII  (1933)
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.
EcoRI  (1)
1 site
G A A T T C C T T A A G
BsaAI  (312)
1 site
Y A C G T R R T G C A Y
NheI  (583)
1 site
G C T A G C C G A T C G
BmtI  (587)
1 site
G C T A G C C G A T C G
AccI  (595)
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.
BstZ17I  (596)
1 site
G T A T A C C A T A T G
XmnI  (639)
1 site
G A A N N N N T T C C T T N N N N A A G
SgrAI  (669)
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.
BsrBI  (757)
1 site
C C G C T C G G C G A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BsrBI will not always regenerate a BsrBI site.
BsrBI is typically used at 37°C, but can be used at temperatures
up to 50°C.
SacII  (835)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
BssSαI  (959)
1 site
C A C G A G G T G C T C
NspI  (1161)
1 site
R C A T G Y Y G T A C R
XbaI  (1424)
1 site
T C T A G A A G A T C T
AcuI  (1484)
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).
KasI  (1705)
1 site
G G C G C C C C G C G G
NarI  (1706)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (1707)
1 site
G G C G C C C C G C G G
PluTI  (1709)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
HincII  (1842)
1 site
G T Y R A C C A R Y T G
HpaI  (1842)
1 site
G T T A A C C A A T T G
BciVI  (1892)
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.
lacI
1617 .. 2699  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
1617 .. 2699  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
CmR
3373 .. 219  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3373 .. 219  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
p15A ori
848 .. 1392  =  545 bp
Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
p15A ori
848 .. 1392  =  545 bp
Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
cat promoter
220 .. 322  =  103 bp
promoter of the E. coli cat gene
cat promoter
220 .. 322  =  103 bp
promoter of the E. coli cat gene
lacI promoter
2700 .. 2777  =  78 bp
lacI promoter
2700 .. 2777  =  78 bp
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