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Plasmid Files

pTV 118N

Bacterial cloning and expression vector with an NcoI site overlapping the lacZα start codon.

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pTV 118N Sequence and MappTV 118N.dna
Map and Sequence File   
Sequence Author:  TaKaRa
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 PspFI (2864) BseYI (2860) AlwNI (2755) AhdI (2276) BsaI (2210) BpmI (2207) NmeAIII (2129) ScaI (1795) XmnI (1676) AflIII - PciI (1) BspQI - SapI (118) M13 Primer RV-N (300 .. 316) BtgI - NcoI - StyI (342) MscI (347) EcoRI (358) Eco53kI (366) SacI (368) Acc65I (370) KpnI - TspMI - XmaI (374) SmaI (376) BamHI (379) XbaI (385) SalI (391) AccI (392) HincII (393) PstI - SbfI (401) BfuAI - BspMI (404) SphI (407) HindIII (409) EcoO109I (631) PfoI (688) PsiI (925) BsaAI - DraIII (1053) BtgZI (1054) NgoMIV (1154) NaeI (1156) pTV 118N 3163 bp
PspFI  (2864)
1 site
C C C A G C G G G T C G
BseYI  (2860)
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  (2755)
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.
AhdI  (2276)
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.
BsaI  (2210)
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.
BpmI  (2207)
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.
NmeAIII  (2129)
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).
ScaI  (1795)
1 site
A G T A C T T C A T G A
XmnI  (1676)
1 site
G A A N N N N T T C C T T N N N N A A G
AflIII  (1)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (1)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (118)
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  (118)
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.
BtgI  (342)
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  (342)
1 site
C C A T G G G G T A C C
StyI  (342)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
MscI  (347)
1 site
T G G C C A A C C G G T
EcoRI  (358)
1 site
G A A T T C C T T A A G
Eco53kI  (366)
1 site
G A G C T C C T C G A G
SacI  (368)
1 site
G A G C T C C T C G A G
Acc65I  (370)
1 site
G G T A C C C C A T G G
KpnI  (374)
1 site
G G T A C C C C A T G G
TspMI  (374)
1 site
C C C G G G G G G C C C
XmaI  (374)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (376)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (379)
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.
XbaI  (385)
1 site
T C T A G A A G A T C T
SalI  (391)
1 site
G T C G A C C A G C T G
AccI  (392)
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  (393)
1 site
G T Y R A C C A R Y T G
PstI  (401)
1 site
C T G C A G G A C G T C
SbfI  (401)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (404)
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  (404)
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.
SphI  (407)
1 site
G C A T G C C G T A C G
HindIII  (409)
1 site
A A G C T T T T C G A A
EcoO109I  (631)
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.
PfoI  (688)
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.
PsiI  (925)
1 site
T T A T A A A A T A T T
BsaAI  (1053)
1 site
Y A C G T R R T G C A Y
DraIII  (1053)
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.
BtgZI  (1054)
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.
NgoMIV  (1154)
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  (1156)
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.
M13 Primer RV-N
17-mer  /  53% GC
1 binding site
300 .. 316  =  17 annealed bases
Tm  =  54°C
AmpR
1489 .. 2349  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1489 .. 1557  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1489 .. 2349  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1558 .. 2349  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1489 .. 2349  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2520 .. 3108  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2520 .. 3108  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
829 .. 1284  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
829 .. 1284  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
lacZα
344 .. 613  =  270 bp
89 amino acids  =  10.2 kDa
Product: LacZα fragment of β-galactosidase
lacZα
344 .. 613  =  270 bp
89 amino acids  =  10.2 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
1384 .. 1488  =  105 bp
AmpR promoter
1384 .. 1488  =  105 bp
lac promoter
269 .. 299  =  31 bp
   Segment 1:  -35  
   269 .. 274  =  6 bp
promoter for the E. coli lac operon
lac promoter
269 .. 299  =  31 bp
   Segment 2:  
   275 .. 292  =  18 bp
promoter for the E. coli lac operon
lac promoter
269 .. 299  =  31 bp
   Segment 3:  -10  
   293 .. 299  =  7 bp
promoter for the E. coli lac operon
lac promoter
269 .. 299  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
307 .. 323  =  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
307 .. 323  =  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).
MCS
358 .. 414  =  57 bp
pUC18/19 multiple cloning site
MCS
358 .. 414  =  57 bp
pUC18/19 multiple cloning site
M13 fwd
418 .. 434  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
418 .. 434  =  17 bp
common sequencing primer, one of multiple similar
variants
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