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Plasmid Files

pSpark® V (linearized)

Linearized transcription-free low copy vector with an advanced MCS for cloning of blunt-ended PCR products containing unstable and toxic genes.

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pSpark V (linearized) Sequence and MappSpark V (linearized).dna
Map and Sequence File   
Sequence Author:  Canvax Biotech
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 T7 promoter NaeI (3009) NgoMIV (3007) BtgZI (2907) DraIII (2906) PsiI (2778) BsaHI (2249) ScaI (2192) AseI (1884) NmeAIII (1860) BsaI (1773) AhdI (1712) PspOMI (3327) EcoO109I (3328) ApaI (3331) BtgI - NcoI - StyI (3333) PmeI (3342) EcoRI (3347) BspEI * (3353) StuI (3363) < EcoRV > (3369) < EcoRV > (0) BamHI (5) SpeI (11) BfuAI - BspMI - NotI (18) PstI - SbfI (29) SalI (31) HincII (33) NdeI (41) Eco53kI (51) AvaI - BsoBI - PaeR7I - PspXI - XhoI (52) BmeT110I - SacI (53) MluI (60) NsiI (73) BspQI - SapI (145) BsmBI (458) PfoI (459) PflFI - Tth111I (562) BstZ17I (588) PciI (819) BseYI (1123) PspFI (1127) AlwNI (1235) pSpark® V 3369 bp
NaeI  (3009)
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.
NgoMIV  (3007)
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.
BtgZI  (2907)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
DraIII  (2906)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
PsiI  (2778)
1 site
T T A T A A A A T A T T
BsaHI  (2249)
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.
ScaI  (2192)
1 site
A G T A C T T C A T G A
AseI  (1884)
1 site
A T T A A T T A A T T A
NmeAIII  (1860)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsaI  (1773)
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  (1712)
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.
PspOMI  (3327)
1 site
G G G C C C C C C G G G
EcoO109I  (3328)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
ApaI  (3331)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BtgI  (3333)
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  (3333)
1 site
C C A T G G G G T A C C
StyI  (3333)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
PmeI  (3342)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (3347)
1 site
G A A T T C C T T A A G
BspEI  (3353)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
StuI  (3363)
1 site
A G G C C T T C C G G A
End  (3369)
0 sites
Start  (0)
0 sites
BamHI  (5)
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.
SpeI  (11)
1 site
A C T A G T T G A T C A
BfuAI  (18)
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  (18)
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.
NotI  (18)
1 site
G C G G C C G C C G C C G G C G
PstI  (29)
1 site
C T G C A G G A C G T C
SbfI  (29)
1 site
C C T G C A G G G G A C G T C C
SalI  (31)
1 site
G T C G A C C A G C T G
HincII  (33)
1 site
G T Y R A C C A R Y T G
NdeI  (41)
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.
Eco53kI  (51)
1 site
G A G C T C C T C G A G
AvaI  (52)
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  (52)
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  (52)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (52)
1 site
V C T C G A G B B G A G C T C V
XhoI  (52)
1 site
C T C G A G G A G C T C
BmeT110I  (53)
1 site
C Y C G R G G R G C Y C
SacI  (53)
1 site
G A G C T C C T C G A G
MluI  (60)
1 site
A C G C G T T G C G C A
NsiI  (73)
1 site
A T G C A T T A C G T A
BspQI  (145)
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  (145)
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.
BsmBI  (458)
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.
PfoI  (459)
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.
PflFI  (562)
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  (562)
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.
BstZ17I  (588)
1 site
G T A T A C C A T A T G
PciI  (819)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (1123)
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.
PspFI  (1127)
1 site
C C C A G C G G G T C G
AlwNI  (1235)
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.
AmpR
1639 .. 2499  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1639 .. 2430  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1639 .. 2499  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2431 .. 2499  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1639 .. 2499  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
880 .. 1468  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
880 .. 1468  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
2682 .. 3137  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2682 .. 3137  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
rop
257 .. 448  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
rop
257 .. 448  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at
low copy number
LacZα
1 .. 126  =  126 bp
42 amino acids  =  4.6 kDa
Product: LacZα fragment of β-galactosidase
LacZα
1 .. 126  =  126 bp
42 amino acids  =  4.6 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
2500 .. 2604  =  105 bp
AmpR promoter
2500 .. 2604  =  105 bp
MCS
3327 .. 3369  =  43 bp
multiple cloning site
MCS
3327 .. 3369  =  43 bp
multiple cloning site
T7 promoter
3301 .. 3319  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3301 .. 3319  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 fwd
3278 .. 3294  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
3278 .. 3294  =  17 bp
common sequencing primer, one of multiple similar
variants
LacZα
3114 .. 3368  =  255 bp
84 amino acids  =  9.7 kDa
Product: LacZα fragment of β-galactosidase
LacZα
3114 .. 3368  =  255 bp
84 amino acids  =  9.7 kDa
Product: LacZα fragment of β-galactosidase
MCS
1 .. 65  =  65 bp
multiple cloning site
MCS
1 .. 65  =  65 bp
multiple cloning site
SP6 promoter
86 .. 104  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
86 .. 104  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 rev
122 .. 138  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
122 .. 138  =  17 bp
common sequencing primer, one of multiple similar
variants
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