pET-54-DEST

Gateway® Nova bacterial destination vector for expressing proteins tagged at the C-terminus with S-Tag.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pET-54-DEST
AvrII (6764) AflII (6751) SalI (6558) PstI (6556) BfuAI - BspMI (6545) SmaI - SrfI (6301) AvaI - BsoBI - TspMI - XmaI (6299) BbvCI (6155) PasI (5662) EcoRI (5430) EagI - NotI (5110) 6xHis ATG NcoI (4955) RBS XbaI (4916) T7 promoter BspDI * - ClaI * (4849) SgrAI (4805) SphI (4657) EcoNI (4592) BstAPI (4448) BclI * (4110) BstEII (3942) ApaI (3921) PspOMI (3917) HpaI (3622) BlpI (6782) DraIII (245) PsiI (370) AhdI (655) BsaI (716) BglI (775) FspI (877) PvuI (1025) PciI (2260) BspQI - SapI (2377) PflFI - Tth111I (2518) PpuMI (3255) pET-54-DEST™ 6865 bp
AvrII  (6764)
1 site		 C C T A G G G G A T C C
AflII  (6751)
1 site		 C T T A A G G A A T T C
SalI  (6558)
1 site		 G T C G A C C A G C T G
PstI  (6556)
1 site		 C T G C A G G A C G T C
BfuAI  (6545)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (6545)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
SmaI  (6301)
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  (6301)
1 site		 G C C C G G G C C G G G C C C G
AvaI  (6299)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (6299)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
TspMI  (6299)
1 site		 C C C G G G G G G C C C
XmaI  (6299)
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.
BbvCI  (6155)
1 site		 C C T C A G C G G A G T C G
PasI  (5662)
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.
EcoRI  (5430)
1 site		 G A A T T C C T T A A G
EagI  (5110)
1 site		 C G G C C G G C C G G C
NotI  (5110)
1 site		 G C G G C C G C C G C C G G C G
NcoI  (4955)
1 site		 C C A T G G G G T A C C
XbaI  (4916)
1 site		 T C T A G A A G A T C T
BspDI  (4849)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (4849)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
SgrAI  (4805)
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.
SphI  (4657)
1 site		 G C A T G C C G T A C G
EcoNI  (4592)
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.
BstAPI  (4448)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
BclI  (4110)
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  (3942)
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  (3921)
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  (3917)
1 site		 G G G C C C C C C G G G
HpaI  (3622)
1 site		 G T T A A C C A A T T G
BlpI  (6782)
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.
DraIII  (245)
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.
PsiI  (370)
1 site		 T T A T A A A A T A T T
AhdI  (655)
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.
BsaI  (716)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BglI  (775)
1 site		 G C C N N N N N G G C C G G N N N N N C C G
Sticky ends from different BglI sites may not be compatible.
FspI  (877)
1 site		 T G C G C A A C G C G T
PvuI  (1025)
1 site		 C G A T C G G C T A G C
PciI  (2260)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (2377)
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  (2377)
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.
PflFI  (2518)
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  (2518)
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.
PpuMI  (3255)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
lacI
3397 .. 4479  =  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
3397 .. 4479  =  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
582 .. 1442  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   582 .. 1373  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
582 .. 1442  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1374 .. 1442  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
582 .. 1442  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
5217 .. 5876  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
5217 .. 5876  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
1616 .. 2204  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1616 .. 2204  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
12 .. 467  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
12 .. 467  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
ccdB
6218 .. 6523  =  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
6218 .. 6523  =  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
2634 .. 2825  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
2634 .. 2825  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
attR1
4984 .. 5108  =  125 bp
recombination site for the Gateway® LR reaction
attR1
4984 .. 5108  =  125 bp
recombination site for the Gateway® LR reaction
attR2
6564 .. 6688  =  125 bp
recombination site for the Gateway® LR reaction
attR2
6564 .. 6688  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
1443 .. 1535  =  93 bp
AmpR promoter
1443 .. 1535  =  93 bp
lacI promoter
4480 .. 4557  =  78 bp
lacI promoter
4480 .. 4557  =  78 bp
T7 terminator
6793 .. 6840  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
6793 .. 6840  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
S-Tag
6693 .. 6737  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
6693 .. 6737  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
lac UV5 promoter
5133 .. 5163  =  31 bp
3 segments
   Segment 1:  -35  
   5133 .. 5138  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
5133 .. 5163  =  31 bp
3 segments
   Segment 2:  
   5139 .. 5156  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
5133 .. 5163  =  31 bp
3 segments
   Segment 3:  -10  
   5157 .. 5163  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
5133 .. 5163  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
lac operator
4889 .. 4913  =  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
4889 .. 4913  =  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).
ATG
4957 .. 4959  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4957 .. 4959  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
4963 .. 4980  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4963 .. 4980  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
T7 promoter
4870 .. 4888  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4870 .. 4888  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
4944 .. 4949  =  6 bp
ribosome binding site
RBS
4944 .. 4949  =  6 bp
ribosome binding site
ORF:  712 .. 978  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2857 .. 3225  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  4655 .. 4906  =  252 bp
ORF:  83 amino acids  =  8.6 kDa
ORF:  6218 .. 6523  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  3363 .. 3614  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  4518 .. 4781  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  5217 .. 5876  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3377 .. 3640  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4229 .. 4729  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
ORF:  3397 .. 4356  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  582 .. 1442  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2634 .. 2858  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
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Individual Sequences & Maps

pACYCDuet-1
pET-24a(+)
pET-3a
pET-51b(+) Ek_LIC (linearized)
pCDF-1b
pET-24b(+)
pET-3b
pET-52b(+)
pCDF-2 Ek_LIC
pET-24c(+)
pET-3c
pET-52b(+) 3C_LIC
pCDF-2 Ek_LIC (linearized)
pET-24d(+)
pET-3d
pET-52b(+) 3C_LIC (linearized)
pCDFDuet-1
pET-25b(+)
pET-40b(+)
pET-53-DEST
pCOLA-2-DEST
pET-26b(+)
pET-41 Ek_LIC
pET-54-DEST
pCOLA-3-DEST
pET-27b(+)
pET-41 Ek_LIC (linearized)
pET-55-DEST
pCOLADuet-1
pET-28a(+)
pET-41a(+)
pET-56-DEST
pET-11a
pET-28b(+)
pET-41b(+)
pET-57-DEST
pET-11b
pET-28c(+)
pET-41c(+)
pET-58-DEST
pET-11c
pET-29a(+)
pET-42a(+)
pET-59-DEST
pET-11d
pET-29b(+)
pET-42b(+)
pET-60-DEST
pET-14b
pET-29c(+)
pET-42c(+)
pET-61-DEST
pET-15b
pET-30 Ek_LIC
pET-43.1 Ek_LIC
pET-62-DEST
pET-16b
pET-30 Ek_LIC (linearized)
pET-43.1 Ek_LIC (linearized)
pET-9a
pET-17b
pET-30 Xa_LIC
pET-43.1a(+)
pET-9b
pET-19b
pET-30 Xa_LIC (linearized)
pET-43.1b(+)
pET-9c
pET-20b(+)
pET-30a(+)
pET-43.1c(+)
pET-9d
pET-21(+)
pET-30b(+)
pET-44a(+)
pETBlue-1
pET-21a(+)
pET-30c(+)
pET-44b(+)
pETBlue-2
pET-21b(+)
pET-31b(+)
pET-44c(+)
pETcoco-1
pET-21c(+)
pET-32 Ek_LIC
pET-45b(+)
pETcoco-2
pET-21d(+)
pET-32 Ek_LIC (linearized)
pET-46 Ek_LIC
pETDuet-1
pET-22b(+)
pET-32 Xa_LIC
pET-46 Ek_LIC (linearized)
pLacI
pET-23(+)
pET-32 Xa_LIC (linearized)
pET-47b(+)
pLysE
pET-23a(+)
pET-32a(+)
pET-48b(+)
pLysS
pET-23b(+)
pET-32b(+)
pET-49b(+)
pRSF-1b
pET-23c(+)
pET-32c(+)
pET-50b(+)
pRSF-2 Ek_LIC
pET-23d(+)
pET-33b(+)
pET-51b(+)
pRSF-2 Ek_LIC (linearized)
pET-24(+)
pET-39b(+)
pET-51b(+) Ek_LIC
pRSFDuet-1

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