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Plasmid Files

pFB-LIC-Bse

Baculovirus 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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pFB-LIC-Bse Sequence and MappFB-LIC-Bse.dna
Map and Sequence File   
Sequence Author:  Structural Genomics Consortium
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 BbsI (6670) Pc promoter PflFI - Tth111I (6141) BsmBI (6095) BstXI (5630) BspQI - SapI (5294) PspFI (4877) BseYI (4873) AlwNI (4768) BpmI (4220) FspI (4066) PvuI (3920) RsrII (136) NcoI (146) 6xHis BseRI (220) NheI (221) BmtI (225) NotI (281) BfuAI - BspMI (310) sacB promoter StuI * (981) BsgI (1305) Bpu10I (1453) SpeI (2227) BseRI (2233) BamHI (2249) EcoRI (2255) Eco53kI (2263) SacI (2265) SalI (2268) HindIII (2274) MfeI (2389) BclI * (2538) AvrII (2553) NgoMIV (3040) NaeI (3042) pFB-LIC-Bse 6816 bp
BbsI  (6670)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
PflFI  (6141)
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  (6141)
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.
BsmBI  (6095)
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.
BstXI  (5630)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BspQI  (5294)
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  (5294)
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.
PspFI  (4877)
1 site
C C C A G C G G G T C G
BseYI  (4873)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
AlwNI  (4768)
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.
BpmI  (4220)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
FspI  (4066)
1 site
T G C G C A A C G C G T
PvuI  (3920)
1 site
C G A T C G G C T A G C
RsrII  (136)
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.
NcoI  (146)
1 site
C C A T G G G G T A C C
BseRI  (220)
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.
NheI  (221)
1 site
G C T A G C C G A T C G
BmtI  (225)
1 site
G C T A G C C G A T C G
NotI  (281)
1 site
G C G G C C G C C G C C G G C G
BfuAI  (310)
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  (310)
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.
StuI  (981)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
BsgI  (1305)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
Bpu10I  (1453)
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.
SpeI  (2227)
1 site
A C T A G T T G A T C A
BseRI  (2233)
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.
BamHI  (2249)
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.
EcoRI  (2255)
1 site
G A A T T C C T T A A G
Eco53kI  (2263)
1 site
G A G C T C C T C G A G
SacI  (2265)
1 site
G A G C T C C T C G A G
SalI  (2268)
1 site
G T C G A C C A G C T G
HindIII  (2274)
1 site
A A G C T T T T C G A A
MfeI  (2389)
1 site
C A A T T G G T T A A C
BclI  (2538)
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.
AvrII  (2553)
1 site
C C T A G G G G A T C C
NgoMIV  (3040)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (3042)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
SacB
740 .. 2161  =  1422 bp
473 amino acids  =  53.0 kDa
   Segment 1:  signal peptide  
   740 .. 826  =  87 bp
   29 amino acids  =  3.0 kDa
Product: secreted levansucrase that renders
bacterial growth sensitive to sucrose
negative selection marker
SacB
740 .. 2161  =  1422 bp
473 amino acids  =  53.0 kDa
   Segment 2:  
   827 .. 2161  =  1335 bp
   444 amino acids  =  50.0 kDa
Product: secreted levansucrase that renders
bacterial growth sensitive to sucrose
negative selection marker
SacB
740 .. 2161  =  1422 bp
473 amino acids  =  53.0 kDa
2 segments
Product: secreted levansucrase that renders
bacterial growth sensitive to sucrose
negative selection marker
AmpR
3502 .. 4362  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3502 .. 3570  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3502 .. 4362  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3571 .. 4362  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3502 .. 4362  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
4533 .. 5121  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4533 .. 5121  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
GmR
5715 .. 6248  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
5715 .. 6248  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
f1 ori
2915 .. 3370  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2915 .. 3370  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
sacB promoter
294 .. 739  =  446 bp
sacB promoter and control region
sacB promoter
294 .. 739  =  446 bp
sacB promoter and control region
Tn7R
5424 .. 5648  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7R
5424 .. 5648  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7L
2566 .. 2731  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
Tn7L
2566 .. 2731  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
SV40 poly(A) signal
2403 .. 2537  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2403 .. 2537  =  135 bp
SV40 polyadenylation signal
AmpR promoter
3397 .. 3501  =  105 bp
AmpR promoter
3397 .. 3501  =  105 bp
polyhedrin promoter
1 .. 92  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
1 .. 92  =  92 bp
promoter for the baculovirus polyhedrin gene
loxP
246 .. 279  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
246 .. 279  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
Pc promoter
6437 .. 6465  =  29 bp
   Segment 3:  -10  
   6437 .. 6442  =  6 bp
class 1 integron promoter
Pc promoter
6437 .. 6465  =  29 bp
   Segment 2:  
   6443 .. 6459  =  17 bp
class 1 integron promoter
Pc promoter
6437 .. 6465  =  29 bp
   Segment 1:  -35  
   6460 .. 6465  =  6 bp
class 1 integron promoter
Pc promoter
6437 .. 6465  =  29 bp
3 segments
class 1 integron promoter
TEV site
196 .. 216  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV site
196 .. 216  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
6xHis
154 .. 171  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
154 .. 171  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
148 .. 150  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
148 .. 150  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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