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pFB-LIC-Bse (linearized)

Linearized baculovirus vector for ligation-independent cloning (LIC), encoding an N-terminal 6xHis-TEV cassette and ampicillin resistance.

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pFB-LIC-Bse (linearized) Sequence and MappFB-LIC-Bse (linearized).dna
Map and Sequence File   
Sequence Author:  Structural Genomics Consortium
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 RsrII (4719) BbsI (4437) PflFI - Tth111I (3908) BsmBI (3862) BstXI (3397) BspQI - SapI (3061) PspFI (2644) BseYI (2640) AlwNI (2535) NcoI (4729) ATG End (4787) Start (0) BamHI (16) EcoRI (22) Eco53kI (30) SacI (32) SalI (35) HindIII (41) MfeI (156) BclI * (305) AvrII (320) NgoMIV (807) NaeI (809) PvuI (1687) FspI (1833) BpmI (1987) pFB-LIC-Bse 4787 bp
RsrII  (4719)
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.
BbsI  (4437)
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  (3908)
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  (3908)
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  (3862)
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  (3397)
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  (3061)
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  (3061)
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  (2644)
1 site
C C C A G C G G G T C G
BseYI  (2640)
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  (2535)
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.
NcoI  (4729)
1 site
C C A T G G G G T A C C
End  (4787)
0 sites
Start  (0)
0 sites
BamHI  (16)
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  (22)
1 site
G A A T T C C T T A A G
Eco53kI  (30)
1 site
G A G C T C C T C G A G
SacI  (32)
1 site
G A G C T C C T C G A G
SalI  (35)
1 site
G T C G A C C A G C T G
HindIII  (41)
1 site
A A G C T T T T C G A A
MfeI  (156)
1 site
C A A T T G G T T A A C
BclI  (305)
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  (320)
1 site
C C T A G G G G A T C C
NgoMIV  (807)
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  (809)
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.
PvuI  (1687)
1 site
C G A T C G G C T A G C
FspI  (1833)
1 site
T G C G C A A C G C G T
BpmI  (1987)
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.
AmpR
1269 .. 2129  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1269 .. 1337  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1269 .. 2129  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1338 .. 2129  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1269 .. 2129  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2300 .. 2888  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2300 .. 2888  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
GmR
3482 .. 4015  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
3482 .. 4015  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
f1 ori
682 .. 1137  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
682 .. 1137  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
Tn7R
3191 .. 3415  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7R
3191 .. 3415  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7L
333 .. 498  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
Tn7L
333 .. 498  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
SV40 poly(A) signal
170 .. 304  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
170 .. 304  =  135 bp
SV40 polyadenylation signal
AmpR promoter
1164 .. 1268  =  105 bp
AmpR promoter
1164 .. 1268  =  105 bp
polyhedrin promoter
4584 .. 4675  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
4584 .. 4675  =  92 bp
promoter for the baculovirus polyhedrin gene
Pc promoter
4204 .. 4232  =  29 bp
   Segment 3:  -10  
   4204 .. 4209  =  6 bp
class 1 integron promoter
Pc promoter
4204 .. 4232  =  29 bp
   Segment 2:  
   4210 .. 4226  =  17 bp
class 1 integron promoter
Pc promoter
4204 .. 4232  =  29 bp
   Segment 1:  -35  
   4227 .. 4232  =  6 bp
class 1 integron promoter
Pc promoter
4204 .. 4232  =  29 bp
3 segments
class 1 integron promoter
TEV site
4779 .. 4799  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV site
4779 .. 4799  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
6xHis
4737 .. 4754  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4737 .. 4754  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
4731 .. 4733  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4731 .. 4733  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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