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Plasmid Files

pLysS

Plasmid for expressing low levels of T7 lysozyme, an inhibitor of T7 RNA polymerase.

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pLysS Sequence and MappLysS.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
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 BsrDI (4874) NcoI (4586) ScaI (4474) BaeGI - Bme1580I (4424) DrdI - PflFI - Tth111I (4343) Bsu36I (4234) BclI * (4183) BsaBI * (4087) AhdI (3850) BmeT110I (3562) AvaI - BsoBI (3561) BfuAI - BspMI (3190) BstAPI (3187) NruI (3110) PspFI (3089) BseYI (3085) EagI (3075) SalI (2787) EcoNI (2762) SphI (2702) KflI (2505) BsrGI (2454) EcoRI (1) BstZ17I (596) XmnI (639) PfoI * (788) SacII (835) XbaI (1424) EarI (1452) BspDI - ClaI (1518) HindIII (1523) EcoRV (1681) PsiI (1934) BsaI (1992) AflII (2057) BciVI (2338) PmeI (2449) pLysS 4886 bp
BsrDI  (4874)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
NcoI  (4586)
1 site
C C A T G G G G T A C C
ScaI  (4474)
1 site
A G T A C T T C A T G A
BaeGI  (4424)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (4424)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
DrdI  (4343)
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  (4343)
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  (4343)
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  (4234)
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.
BclI  (4183)
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  (4087)
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.
AhdI  (3850)
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.
BmeT110I  (3562)
1 site
C Y C G R G G R G C Y C
AvaI  (3561)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (3561)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
BfuAI  (3190)
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  (3190)
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.
BstAPI  (3187)
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.
NruI  (3110)
1 site
T C G C G A A G C G C T
PspFI  (3089)
1 site
C C C A G C G G G T C G
BseYI  (3085)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
EagI  (3075)
1 site
C G G C C G G C C G G C
SalI  (2787)
1 site
G T C G A C C A G C T G
EcoNI  (2762)
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.
SphI  (2702)
1 site
G C A T G C C G T A C G
KflI  (2505)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
BsrGI  (2454)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
EcoRI  (1)
1 site
G A A T T C C T T A A G
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
PfoI  (788)
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.
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.
XbaI  (1424)
1 site
T C T A G A A G A T C T
EarI  (1452)
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.
BspDI  (1518)
1 site
A T C G A T T A G C T A
ClaI  (1518)
1 site
A T C G A T T A G C T A
HindIII  (1523)
1 site
A A G C T T T T C G A A
EcoRV  (1681)
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.
PsiI  (1934)
1 site
T T A T A A A A T A T T
BsaI  (1992)
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.
AflII  (2057)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
BciVI  (2338)
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.
PmeI  (2449)
1 site
G T T T A A A C C A A A T T T G
CmR
4446 .. 219  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
4446 .. 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.
T7 lysozyme
2012 .. 2467  =  456 bp
151 amino acids  =  16.9 kDa
Product: lysozyme from bacteriophage T7
inhibitor of T7 RNA polymerase
T7 lysozyme
2012 .. 2467  =  456 bp
151 amino acids  =  16.9 kDa
Product: lysozyme from bacteriophage T7
inhibitor of T7 RNA polymerase
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
tet promoter
1504 .. 1532  =  29 bp
   Segment 1:  -35  
   1504 .. 1509  =  6 bp
promoter for tetracycline efflux protein gene
tet promoter
1504 .. 1532  =  29 bp
   Segment 2:  
   1510 .. 1526  =  17 bp
promoter for tetracycline efflux protein gene
tet promoter
1504 .. 1532  =  29 bp
   Segment 3:  -10  
   1527 .. 1532  =  6 bp
promoter for tetracycline efflux protein gene
tet promoter
1504 .. 1532  =  29 bp
3 segments
promoter for tetracycline efflux protein gene
T7 Φ3.8 promoter
1981 .. 2014  =  34 bp
weak promoter for bacteriophage T7 RNA
polymerase
T7 Φ3.8 promoter
1981 .. 2014  =  34 bp
weak promoter for bacteriophage T7 RNA
polymerase
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