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Plasmid Files

pTargeT™ (linearized)

Linearized TA cloning vector with 3'-T overhangs for expressing PCR-amplified genes in mammalian cells.

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pTargeT (linearized) Sequence and MappTargeT (linearized).dna
Map and Sequence File   
Sequence Author:  Promega
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 BglII (4382) AlwNI (3961) AhdI (3482) BpmI (3413) ScaI (3001) XmnI (2882) EcoO109I (2502) PfoI (2443) BstXI (2241) BstBI (2202) RsrII (2036) BssHII (1917) PflFI - Tth111I (1638) BbeI (1523) SfoI (1521) NarI (1520) KasI (1519) BsrGI (4483) SpeI (4539) NdeI (4774) SnaBI (4880) AsiSI (5051) Eco53kI (5114) SacI (5116) BmgBI (5536) AanI (5615) EcoRI (5637) BamHI (5643) NheI (5651) BmtI (5655) PaeR7I - XhoI (5657) AflIII - MluI (5663) End (5672) Start (1) TspMI - XmaI (8) SmaI (10) Acc65I (14) KpnI (18) SalI (20) AccI (21) NotI (28) EcoRI (35) pTargeT™ Sequencing Primer (61 .. 84) lac operator AanI (361) HpaI (381) MfeI (390) BsaBI * (483) BspDI * - ClaI * (487) BspEI (492) PmlI (566) DraIII (747) AanI (872) SexAI * (1093) SfiI (1279) BseRI (1322) StuI (1325) AvrII (1326) pTargeT™ 5671 bp
BglII  (4382)
1 site
A G A T C T T C T A G A
AlwNI  (3961)
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  (3482)
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.
BpmI  (3413)
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.
ScaI  (3001)
1 site
A G T A C T T C A T G A
XmnI  (2882)
1 site
G A A N N N N T T C C T T N N N N A A G
EcoO109I  (2502)
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  (2443)
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.
BstXI  (2241)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BstBI  (2202)
1 site
T T C G A A A A G C T T
RsrII  (2036)
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.
BssHII  (1917)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
PflFI  (1638)
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  (1638)
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.
BbeI  (1523)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the BbeI
recognition sequence.
SfoI  (1521)
1 site
G G C G C C C C G C G G
NarI  (1520)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
KasI  (1519)
1 site
G G C G C C C C G C G G
BsrGI  (4483)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SpeI  (4539)
1 site
A C T A G T T G A T C A
NdeI  (4774)
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.
SnaBI  (4880)
1 site
T A C G T A A T G C A T
AsiSI  (5051)
1 site
G C G A T C G C C G C T A G C G
Eco53kI  (5114)
1 site
G A G C T C C T C G A G
SacI  (5116)
1 site
G A G C T C C T C G A G
BmgBI  (5536)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
AanI  (5615)
3 sites
T T A T A A A A T A T T
EcoRI  (5637)
2 sites
G A A T T C C T T A A G
BamHI  (5643)
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.
NheI  (5651)
1 site
G C T A G C C G A T C G
BmtI  (5655)
1 site
G C T A G C C G A T C G
PaeR7I  (5657)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (5657)
1 site
C T C G A G G A G C T C
AflIII  (5663)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (5663)
1 site
A C G C G T T G C G C A
End  (5672)
0 sites
Start  (1)
0 sites
TspMI  (8)
1 site
C C C G G G G G G C C C
XmaI  (8)
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  (10)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (14)
1 site
G G T A C C C C A T G G
KpnI  (18)
1 site
G G T A C C C C A T G G
SalI  (20)
1 site
G T C G A C C A G C T G
AccI  (21)
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.
NotI  (28)
1 site
G C G G C C G C C G C C G G C G
EcoRI  (35)
2 sites
G A A T T C C T T A A G
AanI  (361)
3 sites
T T A T A A A A T A T T
HpaI  (381)
1 site
G T T A A C C A A T T G
MfeI  (390)
1 site
C A A T T G G T T A A C
BsaBI  (483)
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.
BspDI  (487)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (487)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BspEI  (492)
1 site
T C C G G A A G G C C T
PmlI  (566)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
DraIII  (747)
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.
AanI  (872)
3 sites
T T A T A A A A T A T T
SexAI  (1093)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (1279)
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.
BseRI  (1322)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
StuI  (1325)
1 site
A G G C C T T C C G G A
AvrII  (1326)
1 site
C C T A G G G G A T C C
pTargeT™ Sequencing Primer
24-mer  /  38% GC
1 binding site
61 .. 84  =  24 annealed bases
Tm  =  53°C
AmpR
2695 .. 3555  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2695 .. 2763  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2695 .. 3555  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2764 .. 3555  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2695 .. 3555  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
1392 .. 2186  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
1392 .. 2186  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
ori
3726 .. 4314  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3726 .. 4314  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
515 .. 969  =  455 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
515 .. 969  =  455 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
984 .. 1341  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
984 .. 1341  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
4600 .. 4904  =  305 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4600 .. 4904  =  305 bp
human cytomegalovirus immediate early enhancer
lacZα
5438 .. 5671  =  234 bp
77 amino acids  =  9.0 kDa
Product: LacZα fragment of β-galactosidase
lacZα
5438 .. 5671  =  234 bp
77 amino acids  =  9.0 kDa
Product: LacZα fragment of β-galactosidase
CMV promoter
4905 .. 5116  =  212 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
4905 .. 5116  =  212 bp
human cytomegalovirus (CMV) immediate early
promoter
chimeric intron
5277 .. 5409  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
5277 .. 5409  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
260 .. 381  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
260 .. 381  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2590 .. 2694  =  105 bp
AmpR promoter
2590 .. 2694  =  105 bp
lacZα
2 .. 94  =  93 bp
31 amino acids  =  3.3 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2 .. 94  =  93 bp
31 amino acids  =  3.3 kDa
Product: LacZα fragment of β-galactosidase
poly(A) signal
2250 .. 2298  =  49 bp
synthetic polyadenylation signal
poly(A) signal
2250 .. 2298  =  49 bp
synthetic polyadenylation signal
lac promoter
138 .. 168  =  31 bp
   Segment 3:  -10  
   138 .. 144  =  7 bp
promoter for the E. coli lac operon
lac promoter
138 .. 168  =  31 bp
   Segment 2:  
   145 .. 162  =  18 bp
promoter for the E. coli lac operon
lac promoter
138 .. 168  =  31 bp
   Segment 1:  -35  
   163 .. 168  =  6 bp
promoter for the E. coli lac operon
lac promoter
138 .. 168  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
114 .. 130  =  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
114 .. 130  =  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).
SV40 ori
1192 .. 1327  =  136 bp
SV40 origin of replication
SV40 ori
1192 .. 1327  =  136 bp
SV40 origin of replication
MCS
2 .. 40  =  39 bp
multiple cloning site
MCS
2 .. 40  =  39 bp
multiple cloning site
MCS
5637 .. 5671  =  35 bp
multiple cloning site
MCS
5637 .. 5671  =  35 bp
multiple cloning site
T7 promoter
5616 .. 5634  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
5616 .. 5634  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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