pT-REx-DEST30

Gateway® destination vector enabling tetracycline-inducible high-level expression.

Sequence Author: Thermo Fisher (Invitrogen)

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CMV enhancer PciI (7149) AhdI (6261) XmnI (5661) PfoI (5222) BstBI (4967) RsrII (4801) PflFI - Tth111I (4403) PluTI (4288) SfoI (4286) NarI (4285) KasI (4284) AflII (4139) AvrII (4091) StuI (4090) BseRI (4087) SfiI (4044) CsiI - SexAI * (3858) KpnI (3745) SnaBI (288) tet operator tet operator SgrDI (560) BbsI (636) AgeI (674) EcoRV (682) PaeR7I - PspXI - XhoI (686) EcoRI (1145) PasI - PflMI * (1377) BstZ17I (1680) BbvCI (1870) TspMI - XmaI (2014) SmaI - SrfI (2016) BmgBI (2050) PspOMI (2424) ApaI (2428) MluI (2436) T7 promoter NheI (2532) BmtI (2536) BclI * (2808) MfeI (2901) HpaI (2914) DraIII (3408) Acc65I (3741) pT-REx™-DEST30 7544 bp
PciI  (7149)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AhdI  (6261)
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  (5661)
1 site		 G A A N N N N T T C C T T N N N N A A G
PfoI  (5222)
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  (4967)
1 site		 T T C G A A A A G C T T
RsrII  (4801)
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  (4403)
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  (4403)
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  (4288)
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  (4286)
1 site		 G G C G C C C C G C G G
NarI  (4285)
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  (4284)
1 site		 G G C G C C C C G C G G
AflII  (4139)
1 site		 C T T A A G G A A T T C
AvrII  (4091)
1 site		 C C T A G G G G A T C C
StuI  (4090)
1 site		 A G G C C T T C C G G A
BseRI  (4087)
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  (4044)
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.
CsiI  (3858)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (3858)
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  (3745)
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
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  (674)
1 site		 A C C G G T T G G C C A
EcoRV  (682)
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  (686)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (686)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (686)
1 site		 C T C G A G G A G C T C
EcoRI  (1145)
1 site		 G A A T T C C T T A A G
PasI  (1377)
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  (1377)
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  (1680)
1 site		 G T A T A C C A T A T G
BbvCI  (1870)
1 site		 C C T C A G C G G A G T C G
TspMI  (2014)
1 site		 C C C G G G G G G C C C
XmaI  (2014)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2016)
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  (2016)
1 site		 G C C C G G G C C G G G C C C G
BmgBI  (2050)
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.
PspOMI  (2424)
1 site		 G G G C C C C C C G G G
ApaI  (2428)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
MluI  (2436)
1 site		 A C G C G T T G C G C A
NheI  (2532)
1 site		 G C T A G C C G A T C G
BmtI  (2536)
1 site		 G C T A G C C G A T C G
BclI  (2808)
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  (2901)
1 site		 C A A T T G G T T A A C
HpaI  (2914)
1 site		 G T T A A C C A A T T G
DraIII  (3408)
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  (3741)
1 site		 G G T A C C C C A T G G
AmpR
5474 .. 6334  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5474 .. 5542  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5474 .. 6334  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5543 .. 6334  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5474 .. 6334  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
4157 .. 4951  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
4157 .. 4951  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
CmR
932 .. 1591  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
932 .. 1591  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
6505 .. 7093  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6505 .. 7093  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3175 .. 3630  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3175 .. 3630  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
3749 .. 4106  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
3749 .. 4106  =  358 bp
SV40 enhancer and early promoter
ccdB
1933 .. 2238  =  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
1933 .. 2238  =  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
2915 .. 3049  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2915 .. 3049  =  135 bp
SV40 polyadenylation signal
attR1
699 .. 823  =  125 bp
recombination site for the Gateway® LR reaction
attR1
699 .. 823  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2279 .. 2403  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2279 .. 2403  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
5369 .. 5473  =  105 bp
AmpR promoter
5369 .. 5473  =  105 bp
poly(A) signal
5015 .. 5063  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5015 .. 5063  =  49 bp
synthetic polyadenylation signal
lac UV5 promoter
848 .. 878  =  31 bp
3 segments
   Segment 1:  -35  
   848 .. 853  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
848 .. 878  =  31 bp
3 segments
   Segment 2:  
   854 .. 871  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
848 .. 878  =  31 bp
3 segments
   Segment 3:  -10  
   872 .. 878  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
848 .. 878  =  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
2465 .. 2483  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2465 .. 2483  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 fwd
2494 .. 2510  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2494 .. 2510  =  17 bp
common sequencing primer, one of multiple similar variants
SV40 ori
3957 .. 4092  =  136 bp
SV40 origin of replication
SV40 ori
3957 .. 4092  =  136 bp
SV40 origin of replication
ORF:  1933 .. 2238  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  932 .. 1591  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4157 .. 4951  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  5474 .. 6334  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  7493 .. 206  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  4329 .. 4715  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  4466 .. 5002  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  5938 .. 6204  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Individual Sequences & Maps

pAd BLOCK-iT-DEST
pDEST R4-R3 Vector II
pENTR11
pHELLSGATE 4
pAd CMV V5-DEST
pDEST10
pENTR1A
pHELLSGATE 8
pAd PL-DEST
pDEST14
pENTR2B
pIB V5-His-DEST
pBAD-DEST49
pDEST15
pENTR3C
pLenti4 BLOCK-iT-DEST
pcDNA-DEST40
pDEST17
pENTR4
pLenti4 V5-DEST
pcDNA-DEST47
pDEST20
pET-53-DEST
pLenti6.2 C-Lumio V5-DEST
pcDNA-DEST53
pDEST24
pET-54-DEST
pLenti6.2 N-Lumio V5-DEST
pcDNA3.1 nV5-DEST
pDEST26
pET-55-DEST
pLenti6.2 V5-DEST
pcDNA3.2 capTEV-CT V5-DEST
pDEST27
pET-56-DEST
pLenti6.3 TO V5-DEST
pcDNA3.2 capTEV-NT V5-DEST
pDEST8
pET-57-DEST
pLenti6.3 V5-DEST
pcDNA3.2 V5-DEST
pDONR221 P5-P2
pET-58-DEST
pLenti6.4 R4R2 V5-DEST
pcDNA6.2 C-EmGFP-DEST
pDONR P2r-P3
pET-59-DEST
pLenti6 BLOCK-iT-DEST
pcDNA6.2 C-YFP-DEST
pDONR P4-P1r
pET-60-DEST
pLenti6 capTEV-CT-DEST
pcDNA6.2 cGeneBLAzer-DEST
pDONR Zeo
pET-61-DEST
pLenti6 capTEV-NT-DEST1
pcDNA6.2 cLumio-DEST
pDONR201
pET-62-DEST
pLenti6 UbC V5-DEST
pcDNA6.2 EmGFP-Bsd V5-DEST
pDONR207
pET-DEST42
pLenti6 V5-DEST
pcDNA6.2 N-EmGFP-DEST
pDONR221
pET104-DEST
pLenti7.3 V5-DEST
pcDNA6.2 N-YFP-DEST
pDONR221 P1-P4
pET104.1-DEST
pMT-DEST48
pcDNA6.2 nGeneBLAzer-DEST
pDONR221 P1-P5r
pET160-DEST
pMT BioEase-DEST
pcDNA6.2 nLumio-DEST
pDONR221 P3-P2
pET161-DEST
pSCREEN-iT lacZ-DEST
pcDNA6.2 V5-DEST
pDONR221 P4r-P3r
pEXP1-DEST
pT-REx-DEST30
pcDNA6.2 V5-pL-DEST
pDONR221 P5-P4
pEXP2-DEST
pT-REx-DEST31
pCMV SPORT6
pDONR222
pEXP3-DEST
pYES-DEST52
pCMV SPORT6.1
pDONR223
pEXP4-DEST
Reading Frame Cassette A
pCOLA-2-DEST
pEF-DEST51
pHELLSGATE
Reading Frame Cassette B
pCOLA-3-DEST
pEF5 FRT V5-DEST
pHELLSGATE 12
Reading Frame Cassette C.1
pDEST R4-R3
pENTR GeneBLAzer

Plasmid Sets

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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
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Yeast Plasmids
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Mammalian Expression Vectors
Structural Genomics Vectors

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