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Plasmid Files

pT7T3D-PacI

Directional cDNA cloning vector. Also known as pT7T3-Pac, pT7T3Pac, pT7T3D-Pac, or pT7T3D.

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pT7T3D-PacI Sequence and MappT7T3D-PacI.dna
Map and Sequence File   
Sequence Author:  I.M.A.G.E. Cpnsortium
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 NaeI (2769) NgoMIV (2767) BanII (2741) BtgZI (2667) DraIII (2666) PsiI (2548) XmnI (2132) BsaHI (2070) ScaI (2013) TatI (2011) NmeAIII (1681) BpmI (1603) BsaI (1594) AhdI (1533) M13 fwd T3 promoter HindIII (192) EagI - NotI (199) PacI (210) BamHI (217) SnaBI (230) EcoRI (239) AbsI - AvaI - BsoBI - PaeR7I - PspXI - XhoI (250) BmeT110I (251) SfiI (254) T7 promoter lac operator BspQI - SapI (524) AflIII - PciI (640) NspI (644) BseYI (944) PspFI (948) AlwNI (1056) pT7T3D-PacI 2897 bp
NaeI  (2769)
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  (2767)
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.
BanII  (2741)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BtgZI  (2667)
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  (2666)
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  (2548)
1 site
T T A T A A A A T A T T
XmnI  (2132)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (2070)
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  (2013)
1 site
A G T A C T T C A T G A
TatI  (2011)
1 site
W G T A C W W C A T G W
NmeAIII  (1681)
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  (1603)
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  (1594)
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  (1533)
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.
HindIII  (192)
1 site
A A G C T T T T C G A A
EagI  (199)
1 site
C G G C C G G C C G G C
NotI  (199)
1 site
G C G G C C G C C G C C G G C G
PacI  (210)
1 site
T T A A T T A A A A T T A A T T
BamHI  (217)
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.
SnaBI  (230)
1 site
T A C G T A A T G C A T
EcoRI  (239)
1 site
G A A T T C C T T A A G
AbsI  (250)
1 site
C C T C G A G G G G A G C T C C
AvaI  (250)
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  (250)
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  (250)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (250)
1 site
V C T C G A G B B G A G C T C V
XhoI  (250)
1 site
C T C G A G G A G C T C
BmeT110I  (251)
1 site
C Y C G R G G R G C Y C
SfiI  (254)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
BspQI  (524)
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  (524)
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  (640)
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  (640)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (644)
1 site
R C A T G Y Y G T A C R
BseYI  (944)
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  (948)
1 site
C C C A G C G G G T C G
AlwNI  (1056)
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
1460 .. 2320  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1460 .. 2251  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1460 .. 2320  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2252 .. 2320  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1460 .. 2320  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
701 .. 1289  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
701 .. 1289  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
2321 .. 2425  =  105 bp
AmpR promoter
2321 .. 2425  =  105 bp
MCS
192 .. 259  =  68 bp
multiple cloning site
MCS
192 .. 259  =  68 bp
multiple cloning site
lac promoter
347 .. 377  =  31 bp
   Segment 3:  -10  
   347 .. 353  =  7 bp
promoter for the E. coli lac operon
lac promoter
347 .. 377  =  31 bp
   Segment 2:  
   354 .. 371  =  18 bp
promoter for the E. coli lac operon
lac promoter
347 .. 377  =  31 bp
   Segment 1:  -35  
   372 .. 377  =  6 bp
promoter for the E. coli lac operon
lac promoter
347 .. 377  =  31 bp
3 segments
promoter for the E. coli lac operon
T3 promoter
169 .. 187  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
169 .. 187  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T7 promoter
266 .. 284  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
266 .. 284  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 fwd
142 .. 158  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
142 .. 158  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
299 .. 315  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
299 .. 315  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
323 .. 339  =  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
323 .. 339  =  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).
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