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pTZ57R/T

Linearized pTZ57R vector with 3'-ddT overhangs for TA cloning of PCR products with blue/white screening.

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pTZ57R_T.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Fermentas)
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NgoMIV (2364) XmnI (1919) BsaHI (1857) ScaI (1800) TatI (1798) NmeAIII (1468) BpmI (1390) BsaI (1381) AhdI (1320) AlwNI (843) PspFI (735) NaeI (2366) BtgZI (2464) BsaAI (2469) DraIII (2472) PsiI (2597) EcoRI (2852) Eco53kI (2860) SacI (2862) Acc65I (2864) KpnI (2868) NruI (2872) BsmI - NsiI (2880) XbaI (2881) End (2888) Start (1) BamHI (5) AvaI - BsoBI - TspMI - XmaI (9) BmeT110I (10) SmaI (11) PspOMI (12) ApaI (16) SalI (18) AccI (19) HincII (20) PstI (27) StuI (31) SphI (39) HindIII (41) BspQI - SapI (311) AflIII - PciI (427) BseYI (731) pTZ57R/T 2887 bp
NgoMIV  (2364)
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  (1919)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (1857)
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  (1800)
1 site
A G T A C T T C A T G A
TatI  (1798)
1 site
W G T A C W W C A T G W
NmeAIII  (1468)
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  (1390)
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  (1381)
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  (1320)
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  (843)
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.
PspFI  (735)
1 site
C C C A G C G G G T C G
NaeI  (2366)
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  (2464)
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  (2469)
1 site
Y A C G T R R T G C A Y
DraIII  (2472)
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  (2597)
1 site
T T A T A A A A T A T T
EcoRI  (2852)
1 site
G A A T T C C T T A A G
Eco53kI  (2860)
1 site
G A G C T C C T C G A G
SacI  (2862)
1 site
G A G C T C C T C G A G
Acc65I  (2864)
1 site
G G T A C C C C A T G G
KpnI  (2868)
1 site
G G T A C C C C A T G G
NruI  (2872)
1 site
T C G C G A A G C G C T
BsmI  (2880)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
NsiI  (2880)
1 site
A T G C A T T A C G T A
XbaI  (2881)
1 site
T C T A G A A G A T C T
End  (2888)
0 sites
Start  (1)
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.
AvaI  (9)
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  (9)
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.
TspMI  (9)
1 site
C C C G G G G G G C C C
XmaI  (9)
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  (10)
1 site
C Y C G R G G R G C Y C
SmaI  (11)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PspOMI  (12)
1 site
G G G C C C C C C G G G
ApaI  (16)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SalI  (18)
1 site
G T C G A C C A G C T G
AccI  (19)
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  (20)
1 site
G T Y R A C C A R Y T G
PstI  (27)
1 site
C T G C A G G A C G T C
StuI  (31)
1 site
A G G C C T T C C G G A
SphI  (39)
1 site
G C A T G C C G T A C G
HindIII  (41)
1 site
A A G C T T T T C G A A
BspQI  (311)
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  (311)
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  (427)
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  (427)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (731)
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.
AmpR
1247 .. 2107  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1247 .. 2038  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1247 .. 2107  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2039 .. 2107  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1247 .. 2107  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
488 .. 1076  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
488 .. 1076  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2239 .. 2694  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2239 .. 2694  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
2108 .. 2212  =  105 bp
AmpR promoter
2108 .. 2212  =  105 bp
lacZα
2 .. 90  =  89 bp
29 amino acids  =  3.3 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2 .. 90  =  89 bp
29 amino acids  =  3.3 kDa
Product: LacZα fragment of β-galactosidase
MCS
2852 .. 2887  =  36 bp
multiple cloning site
MCS
2852 .. 2887  =  36 bp
multiple cloning site
lac promoter
134 .. 164  =  31 bp
   Segment 3:  -10  
   134 .. 140  =  7 bp
promoter for the E. coli lac operon
lac promoter
134 .. 164  =  31 bp
   Segment 2:  
   141 .. 158  =  18 bp
promoter for the E. coli lac operon
lac promoter
134 .. 164  =  31 bp
   Segment 1:  -35  
   159 .. 164  =  6 bp
promoter for the E. coli lac operon
lac promoter
134 .. 164  =  31 bp
3 segments
promoter for the E. coli lac operon
CAP binding site
179 .. 200  =  22 bp
CAP binding activates transcription in the presence of cAMP.
CAP binding site
179 .. 200  =  22 bp
CAP binding activates transcription in the presence of cAMP.
lac operator
110 .. 126  =  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
110 .. 126  =  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
2835 .. 2851  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2835 .. 2851  =  17 bp
common sequencing primer, one of multiple similar variants
lacZα
2686 .. 2887  =  202 bp
66 amino acids  =  7.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2686 .. 2887  =  202 bp
66 amino acids  =  7.6 kDa
Product: LacZα fragment of β-galactosidase
MCS
2 .. 46  =  45 bp
multiple cloning site
MCS
2 .. 46  =  45 bp
multiple cloning site
T7 promoter
49 .. 67  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
49 .. 67  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
86 .. 102  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
86 .. 102  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1377 .. 1643  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1247 .. 2107  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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