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Plasmid Files

pTZ19R

Phagemid vector constructed by inserting the phage f1 ori and the T7 promoter into pUC19. The f1 ori direction is reversed in pTZ19U.

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pTZ19R Sequence and MappTZ19R.dna
Map and Sequence File   
Sequence Author:  Thermo Scientific (Fermentas)
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 NgoMIV (127) XmnI (2544) BsaHI (2482) ScaI (2425) TatI (2423) NmeAIII (2093) BpmI (2015) BsaI (2006) AhdI (1945) AlwNI (1468) NaeI (129) BtgZI (227) BsaAI (232) DraIII (235) PsiI (360) M13 fwd EcoRI (615) Eco53kI (623) SacI (625) Acc65I (627) AvaI - BsoBI - KpnI - TspMI - XmaI (631) BmeT110I (632) SmaI (633) BamHI (636) XbaI (642) SalI (648) AccI (649) HincII (650) PstI - SbfI (658) BfuAI - BspMI (661) SphI (664) HindIII (666) lac operator CAP binding site BspQI - SapI (936) AflIII - PciI (1052) BseYI (1356) PspFI (1360) pTZ19R 2862 bp
NgoMIV  (127)
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.
XmnI  (2544)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (2482)
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  (2425)
1 site
A G T A C T T C A T G A
TatI  (2423)
1 site
W G T A C W W C A T G W
NmeAIII  (2093)
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).
BpmI  (2015)
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  (2006)
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  (1945)
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  (1468)
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.
NaeI  (129)
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.
BtgZI  (227)
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.
BsaAI  (232)
1 site
Y A C G T R R T G C A Y
DraIII  (235)
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  (360)
1 site
T T A T A A A A T A T T
EcoRI  (615)
1 site
G A A T T C C T T A A G
Eco53kI  (623)
1 site
G A G C T C C T C G A G
SacI  (625)
1 site
G A G C T C C T C G A G
Acc65I  (627)
1 site
G G T A C C C C A T G G
AvaI  (631)
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  (631)
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.
KpnI  (631)
1 site
G G T A C C C C A T G G
TspMI  (631)
1 site
C C C G G G G G G C C C
XmaI  (631)
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.
BmeT110I  (632)
1 site
C Y C G R G G R G C Y C
SmaI  (633)
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  (636)
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  (642)
1 site
T C T A G A A G A T C T
SalI  (648)
1 site
G T C G A C C A G C T G
AccI  (649)
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  (650)
1 site
G T Y R A C C A R Y T G
PstI  (658)
1 site
C T G C A G G A C G T C
SbfI  (658)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (661)
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  (661)
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  (664)
1 site
G C A T G C C G T A C G
HindIII  (666)
1 site
A A G C T T T T C G A A
BspQI  (936)
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  (936)
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.
AflIII  (1052)
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  (1052)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (1356)
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  (1360)
1 site
C C C A G C G G G T C G
AmpR
1872 .. 2732  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1872 .. 2663  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1872 .. 2732  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2664 .. 2732  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1872 .. 2732  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1113 .. 1701  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1113 .. 1701  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
2 .. 457  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2 .. 457  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
AmpR promoter
2733 .. 2837  =  105 bp
AmpR promoter
2733 .. 2837  =  105 bp
MCS
615 .. 671  =  57 bp
pUC18/19 multiple cloning site
MCS
615 .. 671  =  57 bp
pUC18/19 multiple cloning site
lac promoter
759 .. 789  =  31 bp
   Segment 3:  -10  
   759 .. 765  =  7 bp
promoter for the E. coli lac operon
lac promoter
759 .. 789  =  31 bp
   Segment 2:  
   766 .. 783  =  18 bp
promoter for the E. coli lac operon
lac promoter
759 .. 789  =  31 bp
   Segment 1:  -35  
   784 .. 789  =  6 bp
promoter for the E. coli lac operon
lac promoter
759 .. 789  =  31 bp
3 segments
promoter for the E. coli lac operon
CAP binding site
804 .. 825  =  22 bp
CAP binding activates transcription in the presence
of cAMP.
CAP binding site
804 .. 825  =  22 bp
CAP binding activates transcription in the presence
of cAMP.
T7 promoter
674 .. 692  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
674 .. 692  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 fwd
598 .. 614  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
598 .. 614  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
711 .. 727  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
711 .. 727  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
735 .. 751  =  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
735 .. 751  =  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).
LacZα
449 .. 715  =  267 bp
88 amino acids  =  10.1 kDa
Product: LacZα fragment of β-galactosidase
LacZα
449 .. 715  =  267 bp
88 amino acids  =  10.1 kDa
Product: LacZα fragment of β-galactosidase
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