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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.dna
Map and Sequence File:    Download    Open   
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
ORF:  529 .. 915  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2980 .. 3258  =  279 bp
ORF:  92 amino acids  =  10.4 kDa
ORF:  3718 .. 250  =  273 bp
ORF:  90 amino acids  =  10.0 kDa
ORF:  1172 .. 1507  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  357 .. 1151  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  666 .. 1202  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  1509 .. 1772  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  2361 .. 2678  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  1202 .. 1447  =  246 bp
ORF:  81 amino acids  =  8.4 kDa
ORF:  1529 .. 2488  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  2947 .. 3219  =  273 bp
ORF:  90 amino acids  =  9.7 kDa
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