pET160-DEST

Gateway® destination vector for high-level bacterial expression of proteins with an N-terminal tetracysteine (Lumio™) tag.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspET160-DEST
BglII (1) SphI (7249) EcoNI (7184) BclI * (6702) BstEII (6534) ApaI (6513) PspOMI (6509) HpaI (6214) PshAI (5875) NruI (5617) FspAI (5135) PflFI - Tth111I (4370) BsaAI (4364) BspQI - SapI (4229) PciI (4112) T7 promoter XbaI (67) RBS NdeI (107) ATG NsiI (113) 6xHis AleI (128) tetracysteine tag TEV Site NotI (322) lac UV5 promoter EcoRI (642) PasI (874) AgeI (1142) BbvCI (1346) TspMI - XmaI (1490) SmaI - SrfI (1492) BmgBI (1526) SalI (1749) PacI (1889) BlpI (1950) T7 terminator BspDI - ClaI (2198) ZraI (2303) AatII (2305) PvuI (2855) AhdI (3224) pET160-DEST 7437 bp
BglII  (1)
1 site		 A G A T C T T C T A G A
SphI  (7249)
1 site		 G C A T G C C G T A C G
EcoNI  (7184)
1 site		 C C T N N N N N A G G G G A N N N N N T C C
The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BclI  (6702)
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.
BstEII  (6534)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
ApaI  (6513)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PspOMI  (6509)
1 site		 G G G C C C C C C G G G
HpaI  (6214)
1 site		 G T T A A C C A A T T G
PshAI  (5875)
1 site		 G A C N N N N G T C C T G N N N N C A G
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
NruI  (5617)
1 site		 T C G C G A A G C G C T
FspAI  (5135)
1 site		 R T G C G C A Y Y A C G C G T R
PflFI  (4370)
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  (4370)
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.
BsaAI  (4364)
1 site		 Y A C G T R R T G C A Y
BspQI  (4229)
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  (4229)
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.
PciI  (4112)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
XbaI  (67)
1 site		 T C T A G A A G A T C T
NdeI  (107)
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.
NsiI  (113)
1 site		 A T G C A T T A C G T A
AleI  (128)
1 site		 C A C N N N N G T G G T G N N N N C A C
NotI  (322)
1 site		 G C G G C C G C C G C C G G C G
EcoRI  (642)
1 site		 G A A T T C C T T A A G
PasI  (874)
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.
AgeI  (1142)
1 site		 A C C G G T T G G C C A
BbvCI  (1346)
1 site		 C C T C A G C G G A G T C G
TspMI  (1490)
1 site		 C C C G G G G G G C C C
XmaI  (1490)
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  (1492)
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  (1492)
1 site		 G C C C G G G C C G G G C C C G
BmgBI  (1526)
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.
SalI  (1749)
1 site		 G T C G A C C A G C T G
PacI  (1889)
1 site		 T T A A T T A A A A T T A A T T
BlpI  (1950)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
BspDI  (2198)
1 site		 A T C G A T T A G C T A
ClaI  (2198)
1 site		 A T C G A T T A G C T A
ZraI  (2303)
1 site		 G A C G T C C T G C A G
AatII  (2305)
1 site		 G A C G T C C T G C A G
PvuI  (2855)
1 site		 C G A T C G G C T A G C
AhdI  (3224)
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.
lacI
5989 .. 7071  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
lacI
5989 .. 7071  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
AmpR
2437 .. 3297  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   2437 .. 2505  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2437 .. 3297  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   2506 .. 3297  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2437 .. 3297  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
429 .. 1088  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
429 .. 1088  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3468 .. 4056  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3468 .. 4056  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ccdB
1409 .. 1714  =  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
1409 .. 1714  =  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.
rop
4486 .. 4677  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
4486 .. 4677  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
attR1
196 .. 320  =  125 bp
recombination site for the Gateway® LR reaction
attR1
196 .. 320  =  125 bp
recombination site for the Gateway® LR reaction
attR2
1755 .. 1879  =  125 bp
recombination site for the Gateway® LR reaction
attR2
1755 .. 1879  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
2332 .. 2436  =  105 bp
AmpR promoter
2332 .. 2436  =  105 bp
ATG
109 .. 111  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
109 .. 111  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
112 .. 129  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
112 .. 129  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
tetracysteine tag
142 .. 159  =  18 bp
6 amino acids  =  584.8 Da
Product: tetracysteine peptide that binds biarsenical labeling reagents
tetracysteine tag
142 .. 159  =  18 bp
6 amino acids  =  584.8 Da
Product: tetracysteine peptide that binds biarsenical labeling reagents
TEV Site
169 .. 189  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) recognition and cleavage site
TEV Site
169 .. 189  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) recognition and cleavage site
lacI promoter
7072 .. 7149  =  78 bp
lacI promoter
7072 .. 7149  =  78 bp
T7 terminator
1961 .. 2008  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1961 .. 2008  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac UV5 promoter
345 .. 375  =  31 bp
3 segments
   Segment 1:  -35  
   345 .. 350  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
345 .. 375  =  31 bp
3 segments
   Segment 2:  
   351 .. 368  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
345 .. 375  =  31 bp
3 segments
   Segment 3:  -10  
   369 .. 375  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
345 .. 375  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
lac operator
40 .. 64  =  25 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
40 .. 64  =  25 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).
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
94 .. 101  =  8 bp
ribosome binding site
RBS
94 .. 101  =  8 bp
ribosome binding site
ORF:  2437 .. 3297  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1409 .. 1714  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4709 .. 5077  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  7247 .. 49  =  240 bp
ORF:  79 amino acids  =  8.0 kDa
ORF:  429 .. 1088  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  5511 .. 6206  =  696 bp
ORF:  231 amino acids  =  25.4 kDa
ORF:  7110 .. 7373  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  7318 .. 129  =  249 bp
ORF:  82 amino acids  =  8.8 kDa
ORF:  4486 .. 4710  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  5074 .. 5412  =  339 bp
ORF:  112 amino acids  =  12.6 kDa
ORF:  5989 .. 6948  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  2901 .. 3167  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5316 .. 5933  =  618 bp
ORF:  205 amino acids  =  21.8 kDa
ORF:  5969 .. 6232  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  6821 .. 7321  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
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Download pET160-DEST.dna file

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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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