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Plasmid Files

pSpark® III (linearized)

Linearized high copy vector with an advanced MCS and ampicillin and kanamycin resistance markers, for very efficient cloning of blunt-ended PCR products.

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pSpark III (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Canvax Biotech
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PspOMI (3939) DraIII (3518) PsiI (3390) XmnI (2923) ScaI (2804) TatI (2802) BpmI (2394) BsaI (2385) AhdI (2324) AlwNI (1847) EcoO109I (3940) ApaI (3943) BtgI - NcoI (3945) PmeI (3954) EcoRI (3959) BspEI * (3965) StuI (3975) < EcoRV > (3981) < EcoRV > (0) BamHI (5) SpeI (11) NotI (18) PstI - SbfI (29) SalI (31) AccI (32) HincII (33) NdeI (41) Eco53kI (51) AvaI - BsoBI - PaeR7I - PspXI - XhoI (52) BmeT110I - SacI (53) MluI (60) NsiI (73) NruI (344) AgeI (348) XcmI (458) BclI * (483) BsaBI * (501) MscI (724) PflFI - Tth111I (760) SphI (1047) RsrII (1158) PciI (1431) pSpark® III 3981 bp
PspOMI  (3939)
1 site
G G G C C C C C C G G G
DraIII  (3518)
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  (3390)
1 site
T T A T A A A A T A T T
XmnI  (2923)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2804)
1 site
A G T A C T T C A T G A
TatI  (2802)
1 site
W G T A C W W C A T G W
BpmI  (2394)
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.
BsaI  (2385)
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  (2324)
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  (1847)
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.
EcoO109I  (3940)
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  (3943)
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  (3945)
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  (3945)
1 site
C C A T G G G G T A C C
PmeI  (3954)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (3959)
1 site
G A A T T C C T T A A G
BspEI  (3965)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
StuI  (3975)
1 site
A G G C C T T C C G G A
End  (3981)
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
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
AccI  (32)
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.
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
NruI  (344)
1 site
T C G C G A A G C G C T
AgeI  (348)
1 site
A C C G G T T G G C C A
XcmI  (458)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BclI  (483)
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.
BsaBI  (501)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
MscI  (724)
1 site
T G G C C A A C C G G T
PflFI  (760)
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  (760)
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.
SphI  (1047)
1 site
G C A T G C C G T A C G
RsrII  (1158)
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.
PciI  (1431)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
2251 .. 3111  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2251 .. 3042  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2251 .. 3111  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3043 .. 3111  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2251 .. 3111  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
514 .. 1308  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
514 .. 1308  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
1492 .. 2080  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1492 .. 2080  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3294 .. 3749  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3294 .. 3749  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
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
3112 .. 3216  =  105 bp
AmpR promoter
3112 .. 3216  =  105 bp
MCS
3939 .. 3981  =  43 bp
multiple cloning site
MCS
3939 .. 3981  =  43 bp
multiple cloning site
lac promoter
170 .. 200  =  31 bp
   Segment 3:  -10  
   170 .. 176  =  7 bp
promoter for the E. coli lac operon
lac promoter
170 .. 200  =  31 bp
   Segment 2:  
   177 .. 194  =  18 bp
promoter for the E. coli lac operon
lac promoter
170 .. 200  =  31 bp
   Segment 1:  -35  
   195 .. 200  =  6 bp
promoter for the E. coli lac operon
lac promoter
170 .. 200  =  31 bp
3 segments
promoter for the E. coli lac operon
T7 promoter
3913 .. 3931  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3913 .. 3931  =  19 bp
promoter for bacteriophage T7 RNA polymerase
lac operator
146 .. 162  =  17 bp
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).
lac operator
146 .. 162  =  17 bp
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).
M13 fwd
3890 .. 3906  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
3890 .. 3906  =  17 bp
common sequencing primer, one of multiple similar variants
LacZα
3726 .. 3980  =  255 bp
84 amino acids  =  9.7 kDa
Product: LacZα fragment of β-galactosidase
LacZα
3726 .. 3980  =  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
ORF:  514 .. 1308  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  686 .. 1072  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2381 .. 2647  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  823 .. 1077  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  2251 .. 3111  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3726 .. 3980  =  255 bp
ORF:  84 amino acids  =  9.7 kDa  (no start codon)
ORF:  3008 .. 3289  =  282 bp
ORF:  93 amino acids  =  10.9 kDa
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