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Plasmid Files

pT-REx™-DEST31

Gateway® destination vector enabling N-terminal 6xHis-tagging and tetracycline-inducible high-level expression.

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pT-REx-DEST31 Sequence and MappT-REx-DEST31.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 CMV enhancer PciI (7164) AhdI (6276) XmnI (5676) PfoI (5237) BstBI (4982) RsrII (4816) PflFI - Tth111I (4418) PluTI (4303) SfoI (4301) NarI (4300) KasI (4299) AflII (4154) AvrII (4106) StuI (4105) BseRI (4102) SfiI (4059) SexAI * (3873) KpnI (3760) SnaBI (288) tet operator BglII (534) tet operator SgrDI (560) BbsI (636) ATG 6xHis AgeI - SgrAI (691) AleI (692) EcoRV (699) PaeR7I - PspXI - XhoI (703) EcoRI (1161) PasI - PflMI * (1393) BstZ17I (1696) BbvCI (1886) TspMI - XmaI (2030) SmaI - SrfI (2032) BmgBI (2066) XbaI (2433) PspOMI (2439) ApaI (2443) T7 promoter NheI (2547) BmtI (2551) BclI * (2823) MfeI (2916) HpaI (2929) DraIII (3423) Acc65I (3756) pT-REx™-DEST31 7559 bp
PciI  (7164)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (6276)
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.
XmnI  (5676)
1 site
G A A N N N N T T C C T T N N N N A A G
PfoI  (5237)
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.
BstBI  (4982)
1 site
T T C G A A A A G C T T
RsrII  (4816)
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.
PflFI  (4418)
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  (4418)
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.
PluTI  (4303)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (4301)
1 site
G G C G C C C C G C G G
NarI  (4300)
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  (4299)
1 site
G G C G C C C C G C G G
AflII  (4154)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
AvrII  (4106)
1 site
C C T A G G G G A T C C
StuI  (4105)
1 site
A G G C C T T C C G G A
BseRI  (4102)
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.
SfiI  (4059)
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.
SexAI  (3873)
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.
KpnI  (3760)
1 site
G G T A C C C C A T G G
SnaBI  (288)
1 site
T A C G T A A T G C A T
BglII  (534)
1 site
A G A T C T T C T A G A
SgrDI  (560)
1 site
C G T C G A C G G C A G C T G C
BbsI  (636)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
AgeI  (691)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SgrAI  (691)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
AleI  (692)
1 site
C A C N N N N G T G G T G N N N N C A C
EcoRV  (699)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
PaeR7I  (703)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (703)
1 site
V C T C G A G B B G A G C T C V
XhoI  (703)
1 site
C T C G A G G A G C T C
EcoRI  (1161)
1 site
G A A T T C C T T A A G
PasI  (1393)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (1393)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BstZ17I  (1696)
1 site
G T A T A C C A T A T G
BbvCI  (1886)
1 site
C C T C A G C G G A G T C G
TspMI  (2030)
1 site
C C C G G G G G G C C C
XmaI  (2030)
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  (2032)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (2032)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (2066)
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.
XbaI  (2433)
1 site
T C T A G A A G A T C T
PspOMI  (2439)
1 site
G G G C C C C C C G G G
ApaI  (2443)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
NheI  (2547)
1 site
G C T A G C C G A T C G
BmtI  (2551)
1 site
G C T A G C C G A T C G
BclI  (2823)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
MfeI  (2916)
1 site
C A A T T G G T T A A C
HpaI  (2929)
1 site
G T T A A C C A A T T G
DraIII  (3423)
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.
Acc65I  (3756)
1 site
G G T A C C C C A T G G
AmpR
5489 .. 6349  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5489 .. 5557  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5489 .. 6349  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5558 .. 6349  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5489 .. 6349  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
4172 .. 4966  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
4172 .. 4966  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
CmR
948 .. 1607  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
948 .. 1607  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
6520 .. 7108  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6520 .. 7108  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3190 .. 3645  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3190 .. 3645  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
3764 .. 4121  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
3764 .. 4121  =  358 bp
SV40 enhancer and early promoter
ccdB
1949 .. 2254  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
ccdB
1949 .. 2254  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
CMV enhancer
9 .. 312  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
9 .. 312  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
313 .. 516  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
313 .. 516  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
2930 .. 3064  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2930 .. 3064  =  135 bp
SV40 polyadenylation signal
attR1
715 .. 839  =  125 bp
recombination site for the Gateway® LR reaction
attR1
715 .. 839  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2295 .. 2419  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2295 .. 2419  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
5384 .. 5488  =  105 bp
AmpR promoter
5384 .. 5488  =  105 bp
poly(A) signal
5030 .. 5078  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5030 .. 5078  =  49 bp
synthetic polyadenylation signal
lac UV5 promoter
864 .. 894  =  31 bp
   Segment 1:  -35  
   864 .. 869  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
864 .. 894  =  31 bp
   Segment 2:  
   870 .. 887  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
864 .. 894  =  31 bp
   Segment 3:  -10  
   888 .. 894  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
864 .. 894  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
tet operator
518 .. 536  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
518 .. 536  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
539 .. 557  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
539 .. 557  =  19 bp
bacterial operator O2 for the tetR and tetA genes
T7 promoter
2480 .. 2498  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2480 .. 2498  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
673 .. 690  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
673 .. 690  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
M13 fwd
2509 .. 2525  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
2509 .. 2525  =  17 bp
common sequencing primer, one of multiple similar
variants
ATG
661 .. 663  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
661 .. 663  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 ori
3972 .. 4107  =  136 bp
SV40 origin of replication
SV40 ori
3972 .. 4107  =  136 bp
SV40 origin of replication
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