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Plasmid Files

pREP4

Bacterial plasmid with p15A origin and lacI gene for regulating expression from pQE vectors.

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pREP4 Sequence and MappREP4.dna
Map and Sequence File   
Sequence Author:  Qiagen
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 BspDI - ClaI (3736) XbaI (3641) SspI (3637) SacII (3052) SgrAI (2886) XmnI (2856) BstZ17I (2813) BmtI (2804) NheI (2800) AfeI (2799) MauBI (2717) NsiI (2701) PflMI (2681) AclI (2596) ApaLI (2276) MluI (2256) BstEII (2074) ApaI (2053) PspOMI (2049) ApoI (1981) HindIII (1) BsaBI * (320) BglII (321) EagI (391) DrdI (512) MscI (567) FspI (587) NcoI (917) RsrII (1001) AvaI - BsoBI - TspMI - XmaI (1322) BmeT110I (1323) SmaI (1324) NruI (1340) Bsu36I (1361) BsmBI (1641) HpaI (1754) EcoRV (1810) pREP4 3740 bp
BspDI  (3736)
1 site
A T C G A T T A G C T A
ClaI  (3736)
1 site
A T C G A T T A G C T A
XbaI  (3641)
1 site
T C T A G A A G A T C T
SspI  (3637)
1 site
A A T A T T T T A T A A
SacII  (3052)
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.
SgrAI  (2886)
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.
XmnI  (2856)
1 site
G A A N N N N T T C C T T N N N N A A G
BstZ17I  (2813)
1 site
G T A T A C C A T A T G
BmtI  (2804)
1 site
G C T A G C C G A T C G
NheI  (2800)
1 site
G C T A G C C G A T C G
AfeI  (2799)
1 site
A G C G C T T C G C G A
MauBI  (2717)
1 site
C G C G C G C G G C G C G C G C
NsiI  (2701)
1 site
A T G C A T T A C G T A
PflMI  (2681)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
AclI  (2596)
1 site
A A C G T T T T G C A A
ApaLI  (2276)
1 site
G T G C A C C A C G T G
MluI  (2256)
1 site
A C G C G T T G C G C A
BstEII  (2074)
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.
ApaI  (2053)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (2049)
1 site
G G G C C C C C C G G G
ApoI  (1981)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
HindIII  (1)
1 site
A A G C T T T T C G A A
BsaBI  (320)
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.
BglII  (321)
1 site
A G A T C T T C T A G A
EagI  (391)
1 site
C G G C C G G C C G G C
DrdI  (512)
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.
MscI  (567)
1 site
T G G C C A A C C G G T
FspI  (587)
1 site
T G C G C A A C G C G T
NcoI  (917)
1 site
C C A T G G G G T A C C
RsrII  (1001)
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.
AvaI  (1322)
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  (1322)
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.
TspMI  (1322)
1 site
C C C G G G G G G C C C
XmaI  (1322)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
BmeT110I  (1323)
1 site
C Y C G R G G R G C Y C
SmaI  (1324)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
NruI  (1340)
1 site
T C G C G A A G C G C T
Bsu36I  (1361)
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.
BsmBI  (1641)
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.
HpaI  (1754)
1 site
G T T A A C C A A T T G
EcoRV  (1810)
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.
lacI
1529 .. 2611  =  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
1529 .. 2611  =  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).
NeoR/KanR
357 .. 1151  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
357 .. 1151  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
p15A ori
3065 .. 3609  =  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
3065 .. 3609  =  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.
lacI promoter
2612 .. 2689  =  78 bp
lacI promoter
2612 .. 2689  =  78 bp
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