pETBlue-1

Vector for blue/white screening and for inducible bacterial expression of native unfused proteins.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pETBlue-1
AgeI (3422) NspI (3090) AflIII - PciI (3086) PspFI (2786) BseYI (2782) AlwNI (2677) BspHI (2366) XmnI (2161) ScaI (2042) TatI (2040) T7 promoter XbaI (47) BglII (223) NheI (235) BmtI (239) EcoRV (278) EcoRI (282) AvaI - BsoBI - TspMI - XmaI (291) SmaI - SrfI (293) tet promoter AvrII (420) BlpI (824) BspEI (901) BanII (1078) BtgZI (1144) BsaAI (1149) DraIII (1152) PsiI (1277) SspI (1357) AhdI (1562) NmeAIII (1710) pETBlue™-1 3476 bp
AgeI  (3422)
1 site		 A C C G G T T G G C C A
NspI  (3090)
1 site		 R C A T G Y Y G T A C R
AflIII  (3086)
1 site		 A C R Y G T T G Y R C A
Sticky ends from different AflIII sites may not be compatible.
PciI  (3086)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
PspFI  (2786)
1 site		 C C C A G C G G G T C G
BseYI  (2782)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AlwNI  (2677)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
BspHI  (2366)
1 site		 T C A T G A A G T A C T
XmnI  (2161)
1 site		 G A A N N N N T T C C T T N N N N A A G
ScaI  (2042)
1 site		 A G T A C T T C A T G A
TatI  (2040)
1 site		 W G T A C W W C A T G W
XbaI  (47)
1 site		 T C T A G A A G A T C T
BglII  (223)
1 site		 A G A T C T T C T A G A
NheI  (235)
1 site		 G C T A G C C G A T C G
BmtI  (239)
1 site		 G C T A G C C G A T C G
EcoRV  (278)
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.
EcoRI  (282)
1 site		 G A A T T C C T T A A G
AvaI  (291)
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  (291)
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  (291)
1 site		 C C C G G G G G G C C C
XmaI  (291)
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  (293)
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  (293)
1 site		 G C C C G G G C C G G G C C C G
AvrII  (420)
1 site		 C C T A G G G G A T C C
BlpI  (824)
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.
BspEI  (901)
1 site		 T C C G G A A G G C C T
BanII  (1078)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
BtgZI  (1144)
1 site		 G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4
Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BsaAI  (1149)
1 site		 Y A C G T R R T G C A Y
DraIII  (1152)
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  (1277)
1 site		 T T A T A A A A T A T T
SspI  (1357)
1 site		 A A T A T T T T A T A A
AhdI  (1562)
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.
NmeAIII  (1710)
1 site		 G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19
Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AmpR
1489 .. 2349  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1489 .. 2280  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1489 .. 2349  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2281 .. 2349  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1489 .. 2349  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2442 .. 3030  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2442 .. 3030  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
919 .. 1374  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
919 .. 1374  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
lacZα
54 .. 314  =  261 bp
86 amino acids  =  10.0 kDa
Product: LacZα fragment of β-galactosidase
lacZα
54 .. 314  =  261 bp
86 amino acids  =  10.0 kDa
Product: LacZα fragment of β-galactosidase
rrnB T1 terminator
562 .. 648  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
562 .. 648  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
835 .. 882  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
835 .. 882  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
tet promoter
364 .. 392  =  29 bp
3 segments
   Segment 3:  -10  
   364 .. 369  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
364 .. 392  =  29 bp
3 segments
   Segment 2:  
   370 .. 386  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
364 .. 392  =  29 bp
3 segments
   Segment 1:  -35  
   387 .. 392  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
364 .. 392  =  29 bp
3 segments
E. coli promoter for tetracycline efflux protein gene
rrnB T2 terminator
740 .. 767  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
740 .. 767  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
lac operator
20 .. 44  =  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
20 .. 44  =  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 (symmetric)
3429 .. 3448  =  20 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). The symmetric lac operator was optimized for tight binding of lac repressor.
lac operator (symmetric)
3429 .. 3448  =  20 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). The symmetric lac operator was optimized for tight binding of lac repressor.
T7 promoter
1 .. 19  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1 .. 19  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
265 .. 270  =  6 bp
ribosome binding site
RBS
265 .. 270  =  6 bp
ribosome binding site
ORF:  1619 .. 1885  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  54 .. 314  =  261 bp
ORF:  86 amino acids  =  10.0 kDa
ORF:  3190 .. 73  =  360 bp
ORF:  119 amino acids  =  12.8 kDa
ORF:  421 .. 729  =  309 bp
ORF:  102 amino acids  =  11.4 kDa
ORF:  1489 .. 2349  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pETBlue-1.dna file

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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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