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Plasmid Files

pLIC-SGC1

Bacterial vector encoding an N-terminal 6xHis-TEV cassette and ampicillin resistance plus a SacB negative selection marker, for purification of recombinant proteins.

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pLIC-SGC1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Structural Genomics Consortium
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BlpI (7087) BmeT110I (7009) AvaI - BsoBI - PaeR7I - PspXI - XhoI (7008) HindIII (6993) SalI (6987) SacI (6984) Eco53kI (6982) EcoRI (6974) BamHI (6968) BseRI (6952) SpeI (6946) SacII (6538) SnaBI (6208) BsrGI (6139) AanI - PsiI (6099) MscI (5722) StuI * (5700) BfuAI - BspMI (5029) BmtI (4944) NheI (4940) BseRI (4939) BglII (4902) NdeI (4868) XbaI (4828) SgrAI (4717) SphI (4569) EcoNI (4504) PflMI (4461) BstAPI (4360) MluI (4036) BclI * (4022) BstEII (3854) ApaI (3833) PspOMI (3829) BssHII (3625) ScaI (566) PvuI (678) PstI (805) BsaI (981) AhdI (1047) AlwNI (1526) BspQI - SapI (2052) PflFI - Tth111I (2193) PpuMI (2930) FspAI (2958) PshAI (3195) pLIC-SGC1 7170 bp
BlpI  (7087)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BmeT110I  (7009)
1 site
C Y C G R G G R G C Y C
AvaI  (7008)
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  (7008)
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.
PaeR7I  (7008)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (7008)
1 site
V C T C G A G B B G A G C T C V
XhoI  (7008)
1 site
C T C G A G G A G C T C
HindIII  (6993)
1 site
A A G C T T T T C G A A
SalI  (6987)
1 site
G T C G A C C A G C T G
SacI  (6984)
1 site
G A G C T C C T C G A G
Eco53kI  (6982)
1 site
G A G C T C C T C G A G
EcoRI  (6974)
1 site
G A A T T C C T T A A G
BamHI  (6968)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BseRI  (6952)
2 sites
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.
SpeI  (6946)
1 site
A C T A G T T G A T C A
SacII  (6538)
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.
SnaBI  (6208)
1 site
T A C G T A A T G C A T
BsrGI  (6139)
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.
AanI  (6099)
1 site
T T A T A A A A T A T T
PsiI  (6099)
1 site
T T A T A A A A T A T T
MscI  (5722)
1 site
T G G C C A A C C G G T
StuI  (5700)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BfuAI  (5029)
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  (5029)
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.
BmtI  (4944)
1 site
G C T A G C C G A T C G
NheI  (4940)
1 site
G C T A G C C G A T C G
BseRI  (4939)
2 sites
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.
BglII  (4902)
1 site
A G A T C T T C T A G A
NdeI  (4868)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
XbaI  (4828)
1 site
T C T A G A A G A T C T
SgrAI  (4717)
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.
SphI  (4569)
1 site
G C A T G C C G T A C G
EcoNI  (4504)
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.
PflMI  (4461)
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.
BstAPI  (4360)
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.
MluI  (4036)
1 site
A C G C G T T G C G C A
BclI  (4022)
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.
BstEII  (3854)
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  (3833)
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  (3829)
1 site
G G G C C C C C C G G G
BssHII  (3625)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
ScaI  (566)
1 site
A G T A C T T C A T G A
PvuI  (678)
1 site
C G A T C G G C T A G C
PstI  (805)
1 site
C T G C A G G A C G T C
BsaI  (981)
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.
AhdI  (1047)
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.
AlwNI  (1526)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
BspQI  (2052)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (2052)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PflFI  (2193)
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  (2193)
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.
PpuMI  (2930)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
FspAI  (2958)
1 site
R T G C G C A Y Y A C G C G T R
PshAI  (3195)
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.
SacB
5459 .. 6880  =  1422 bp
473 amino acids  =  53.0 kDa
   Segment 1:  signal peptide  
   5459 .. 5545  =  87 bp
   29 amino acids  =  3.0 kDa
Product: secreted levansucrase that renders bacterial growth sensitive to sucrose
negative selection marker
SacB
5459 .. 6880  =  1422 bp
473 amino acids  =  53.0 kDa
   Segment 2:  
   5546 .. 6880  =  1335 bp
   444 amino acids  =  50.0 kDa
Product: secreted levansucrase that renders bacterial growth sensitive to sucrose
negative selection marker
SacB
5459 .. 6880  =  1422 bp
473 amino acids  =  53.0 kDa
2 segments
Product: secreted levansucrase that renders bacterial growth sensitive to sucrose
negative selection marker
lacI
3309 .. 4391  =  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
3309 .. 4391  =  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).
AmpR
260 .. 1120  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   260 .. 331  =  72 bp
   24 amino acids &#