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Plasmid Files

pJW168

Bacterial vector allowing inducible expression of Cre recombinase for in vivo site-specific recombination.

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pJW168 Sequence and MappJW168.dna
Map and Sequence File   
Sequence Author:  Lucigen
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 SmaI (5720) BmeT110I (5719) AvaI - BsoBI - TspMI - XmaI (5718) HindIII (5691) BmtI (5674) NheI (5670) BtgI - NcoI - StyI (5664) SpeI (4540) BseRI (4298) AflII (4249) EagI - NotI (4220) AhdI (3647) BsaI (3581) BglI (3529) FspI (3424) ScaI (3166) BssS α I (2978) EcoRI (5723) PshAI (199) Bpu10I (208) AsiSI (578) SexAI * (602) BstBI (623) RsrII (730) NruI (820) AgeI (989) Acc65I (1042) KpnI (1046) PstI (1051) XbaI (1053) StuI (1065) lac operator lac operator PfoI (1307) DrdI (1358) PflFI - Tth111I (1410) PflMI (1451) MluI (1869) BstEII (2050) PspOMI (2076) ApaI - BanII (2080) HpaI (2375) pJW168 5728 bp
SmaI  (5720)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BmeT110I  (5719)
1 site
C Y C G R G G R G C Y C
AvaI  (5718)
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  (5718)
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  (5718)
1 site
C C C G G G G G G C C C
XmaI  (5718)
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.
HindIII  (5691)
1 site
A A G C T T T T C G A A
BmtI  (5674)
1 site
G C T A G C C G A T C G
NheI  (5670)
1 site
G C T A G C C G A T C G
BtgI  (5664)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (5664)
1 site
C C A T G G G G T A C C
StyI  (5664)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
SpeI  (4540)
1 site
A C T A G T T G A T C A
BseRI  (4298)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
AflII  (4249)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
EagI  (4220)
1 site
C G G C C G G C C G G C
NotI  (4220)
1 site
G C G G C C G C C G C C G G C G
AhdI  (3647)
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.
BsaI  (3581)
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.
BglI  (3529)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
FspI  (3424)
1 site
T G C G C A A C G C G T
ScaI  (3166)
1 site
A G T A C T T C A T G A
BssSαI  (2978)
1 site
C A C G A G G T G C T C
EcoRI  (5723)
1 site
G A A T T C C T T A A G
PshAI  (199)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
Bpu10I  (208)
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.
AsiSI  (578)
1 site
G C G A T C G C C G C T A G C G
SexAI  (602)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BstBI  (623)
1 site
T T C G A A A A G C T T
RsrII  (730)
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.
NruI  (820)
1 site
T C G C G A A G C G C T
AgeI  (989)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
Acc65I  (1042)
1 site
G G T A C C C C A T G G
KpnI  (1046)
1 site
G G T A C C C C A T G G
PstI  (1051)
1 site
C T G C A G G A C G T C
XbaI  (1053)
1 site
T C T A G A A G A T C T
StuI  (1065)
1 site
A G G C C T T C C G G A
PfoI  (1307)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
DrdI  (1358)
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  (1410)
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  (1410)
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.
PflMI  (1451)
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.
MluI  (1869)
1 site
A C G C G T T G C G C A
BstEII  (2050)
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.
PspOMI  (2076)
1 site
G G G C C C C C C G G G
ApaI  (2080)
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  (2080)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
HpaI  (2375)
1 site
G T T A A C C A A T T G
lacI
1519 .. 2601  =  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
1519 .. 2601  =  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).
Cre
4 .. 1035  =  1032 bp
343 amino acids  =  38.5 kDa
Product: site-specific recombinase
Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP
sequences.
Cre
4 .. 1035  =  1032 bp
343 amino acids  =  38.5 kDa
Product: site-specific recombinase
Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP
sequences.
Rep101(Ts)
4612 .. 5562  =  951 bp
316 amino acids  =  37.2 kDa
Product: temperature-sensitive version of a protein
needed for replication with the pSC101 origin
Rep101(Ts)
4612 .. 5562  =  951 bp
316 amino acids  =  37.2 kDa
Product: temperature-sensitive version of a protein
needed for replication with the pSC101 origin
AmpR
2860 .. 3720  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2860 .. 2928  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2860 .. 3720  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2929 .. 3720  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2860 .. 3720  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
pSC101 ori
4342 .. 4564  =  223 bp
low-copy replication origin that requires the Rep101
protein
pSC101 ori
4342 .. 4564  =  223 bp
low-copy replication origin that requires the Rep101
protein
AmpR promoter
2755 .. 2859  =  105 bp
AmpR promoter
2755 .. 2859  =  105 bp
lacIq promoter
1441 .. 1518  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
lacIq promoter
1441 .. 1518  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
lac UV5 promoter
1108 .. 1138  =  31 bp
   Segment 3:  -10  
   1108 .. 1114  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1108 .. 1138  =  31 bp
   Segment 2:  
   1115 .. 1132  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1108 .. 1138  =  31 bp
   Segment 1:  -35  
   1133 .. 1138  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1108 .. 1138  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1203 .. 1233  =  31 bp
   Segment 3:  -10  
   1203 .. 1209  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1203 .. 1233  =  31 bp
   Segment 2:  
   1210 .. 1227  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1203 .. 1233  =  31 bp
   Segment 1:  -35  
   1228 .. 1233  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1203 .. 1233  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
lac operator
1084 .. 1100  =  17 bp
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).
lac operator
1084 .. 1100  =  17 bp
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).
lac operator
1179 .. 1195  =  17 bp
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).
lac operator
1179 .. 1195  =  17 bp
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).
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