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Plasmid Files

pFastBac™/CT-TOPO® (linearized)

Linearized baculovirus expression vector for TOPO® cloning of blunt PCR products and addition of a cleavable C-terminal 6xHis tag.

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pFastBac CT-TOPO (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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BstZ17I (4590) AccI (4589) SnaBI (4587) BbsI (4463) BsrGI (4116) PflFI - Tth111I (3934) BsmBI (3888) BseRI (3667) EcoRV (3531) SacII (3474) BstXI (3423) MscI (3415) BspQI - SapI (3085) PspFI (2668) BseYI (2664) AlwNI (2559) BamHI (4738) AgeI (4744) End (4754) Start (0) PaeR7I - PspXI - XhoI (49) SphI (59) Acc65I (61) KpnI (65) HindIII (67) MfeI (182) HpaI (195) BclI * (331) AvrII (346) NgoMIV (833) NaeI (835) BanII (867) BsaHI (1540) PvuI (1711) FspI (1857) BpmI (2011) pFastBac™/CT-TOPO® 4754 bp
BstZ17I  (4590)
1 site
G T A T A C C A T A T G
AccI  (4589)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
SnaBI  (4587)
1 site
T A C G T A A T G C A T
BbsI  (4463)
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.
BsrGI  (4116)
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.
PflFI  (3934)
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  (3934)
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  (3888)
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.
BseRI  (3667)
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.
EcoRV  (3531)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
SacII  (3474)
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.
BstXI  (3423)
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.
MscI  (3415)
1 site
T G G C C A A C C G G T
BspQI  (3085)
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  (3085)
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  (2668)
1 site
C C C A G C G G G T C G
BseYI  (2664)
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  (2559)
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.
BamHI  (4738)
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.
AgeI  (4744)
1 site
A C C G G T T G G C C A
End  (4754)
0 sites
Start  (0)
0 sites
PaeR7I  (49)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (49)
1 site
V C T C G A G B B G A G C T C V
XhoI  (49)
1 site
C T C G A G G A G C T C
SphI  (59)
1 site
G C A T G C C G T A C G
Acc65I  (61)
1 site
G G T A C C C C A T G G
KpnI  (65)
1 site
G G T A C C C C A T G G
HindIII  (67)
1 site
A A G C T T T T C G A A
MfeI  (182)
1 site
C A A T T G G T T A A C
HpaI  (195)
1 site
G T T A A C C A A T T G
BclI  (331)
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  (346)
1 site
C C T A G G G G A T C C
NgoMIV  (833)
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  (835)
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.
BanII  (867)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BsaHI  (1540)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
PvuI  (1711)
1 site
C G A T C G G C T A G C
FspI  (1857)
1 site
T G C G C A A C G C G T
BpmI  (2011)
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
1293 .. 2153  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1293 .. 1361  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1293 .. 2153  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1362 .. 2153  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1293 .. 2153  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2324 .. 2912  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2324 .. 2912  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
GmR
3508 .. 4041  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
3508 .. 4041  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
f1 ori
708 .. 1162  =  455 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
708 .. 1162  =  455 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
Tn7R
3217 .. 3441  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7R
3217 .. 3441  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7L
359 .. 524  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
Tn7L
359 .. 524  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
SV40 poly(A) signal
196 .. 330  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
196 .. 330  =  135 bp
SV40 polyadenylation signal
AmpR promoter
1188 .. 1292  =  105 bp
AmpR promoter
1188 .. 1292  =  105 bp
polyhedrin promoter
4610 .. 4701  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
4610 .. 4701  =  92 bp
promoter for the baculovirus polyhedrin gene
Pc promoter
4230 .. 4258  =  29 bp
   Segment 3:  -10  
   4230 .. 4235  =  6 bp
class 1 integron promoter
Pc promoter
4230 .. 4258  =  29 bp
   Segment 2:  
   4236 .. 4252  =  17 bp
class 1 integron promoter
Pc promoter
4230 .. 4258  =  29 bp
   Segment 1:  -35  
   4253 .. 4258  =  6 bp
class 1 integron promoter
Pc promoter
4230 .. 4258  =  29 bp
3 segments
class 1 integron promoter
TEV site
7 .. 27  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
7 .. 27  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
6xHis
28 .. 45  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
28 .. 45  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ORF:  3646 .. 3978  =  333 bp
ORF:  110 amino acids  =  12.0 kDa
ORF:  4252 .. 4494  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  1293 .. 2153  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3351 .. 3653  =  303 bp
ORF:  100 amino acids  =  11.8 kDa
ORF:  4143 .. 4622  =  480 bp
ORF:  159 amino acids  =  18.5 kDa
ORF:  4233 .. 4541  =  309 bp
ORF:  102 amino acids  =  11.4 kDa
ORF:  1757 .. 2023  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3508 .. 4041  =  534 bp
ORF:  177 amino acids  =  19.4 kDa
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