pJW168

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

Sequence Author: Lucigen

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SmaI (5720) 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) BssSI - BssSαI (2978) EcoRI (5723) PshAI (199) Bpu10I (208) AsiSI (578) CsiI - 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.
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

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
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
BssSI  (2978)
1 site
C A C G A G G T G C T C
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

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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
CsiI  (602)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
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
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  =  ~37.3 kDa
Product: temperature-sensitive version of a protein needed for replication with the pSC101 origin
Rep101(Ts)
4612 .. 5562  =  951 bp
316 amino acids  =  ~37.3 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
2 segments
   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
2 segments
   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