pT-REx-DEST31

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

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspT-REx-DEST31
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) CsiI - SexAI * (3873) KpnI (3760) SnaBI (288) tet operator BglII (534) SgrDI (560) BbsI (636) 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) M13 fwd 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
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.
CsiI  (3873)
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  (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
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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
2 segments
   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
2 segments
   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
3 segments
   Segment 1:  -35  
   864 .. 869  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
864 .. 894  =  31 bp
3 segments
   Segment 2:  
   870 .. 887  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
864 .. 894  =  31 bp
3 segments
   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
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
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
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
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
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
SV40 ori
3972 .. 4107  =  136 bp
SV40 origin of replication
SV40 ori
3972 .. 4107  =  136 bp
SV40 origin of replication
ORF:  472 .. 750  =  279 bp
ORF:  92 amino acids  =  11.3 kDa
ORF:  1949 .. 2254  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4172 .. 4966  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  5489 .. 6349  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  7508 .. 206  =  258 bp
ORF:  85 amino acids  =  9.6 kDa
ORF:  948 .. 1607  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4344 .. 4730  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  4481 .. 5017  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  5953 .. 6219  =  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

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