pETBlue-1 AccepTor

Linearized vector with 3'-U overhangs for UA cloning and inducible bacterial expression of PCR-amplified genes.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsTA and GC Cloning VectorspETBlue-1 AccepTor
AgeI (3145) NspI (2813) AflIII - PciI (2809) PspFI (2509) BseYI (2505) AlwNI (2400) BspHI (2089) XmnI (1884) ScaI (1765) TatI (1763) NmeAIII (1433) T7 promoter XbaI (3246) BglII (3422) NheI (3434) BmtI (3438) End (3478) Start (1) EcoRI (5) AvaI - BsoBI - TspMI - XmaI (14) SmaI - SrfI (16) tet promoter AvrII (143) BlpI (547) BspEI (624) BanII (801) BtgZI (867) BsaAI (872) DraIII (875) PsiI (1000) SspI (1080) AhdI (1285) pETBlue™-1 AccepTor™ 3477 bp
AgeI  (3145)
1 site		 A C C G G T T G G C C A
NspI  (2813)
1 site		 R C A T G Y Y G T A C R
AflIII  (2809)
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  (2809)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
PspFI  (2509)
1 site		 C C C A G C G G G T C G
BseYI  (2505)
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  (2400)
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  (2089)
1 site		 T C A T G A A G T A C T
XmnI  (1884)
1 site		 G A A N N N N T T C C T T N N N N A A G
ScaI  (1765)
1 site		 A G T A C T T C A T G A
TatI  (1763)
1 site		 W G T A C W W C A T G W
NmeAIII  (1433)
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).
XbaI  (3246)
1 site		 T C T A G A A G A T C T
BglII  (3422)
1 site		 A G A T C T T C T A G A
NheI  (3434)
1 site		 G C T A G C C G A T C G
BmtI  (3438)
1 site		 G C T A G C C G A T C G
End  (3478)
0 sites
Start  (1)
0 sites
EcoRI  (5)
1 site		 G A A T T C C T T A A G
AvaI  (14)
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  (14)
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  (14)
1 site		 C C C G G G G G G C C C
XmaI  (14)
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  (16)
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  (16)
1 site		 G C C C G G G C C G G G C C C G
AvrII  (143)
1 site		 C C T A G G G G A T C C
BlpI  (547)
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  (624)
1 site		 T C C G G A A G G C C T
BanII  (801)
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  (867)
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  (872)
1 site		 Y A C G T R R T G C A Y
DraIII  (875)
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  (1000)
1 site		 T T A T A A A A T A T T
SspI  (1080)
1 site		 A A T A T T T T A T A A
AhdI  (1285)
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.
AmpR
1212 .. 2072  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1212 .. 2003  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1212 .. 2072  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2004 .. 2072  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1212 .. 2072  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2165 .. 2753  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2165 .. 2753  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
642 .. 1097  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
642 .. 1097  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
lacZα
3253 .. 3477  =  225 bp
74 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
lacZα
3253 .. 3477  =  225 bp
74 amino acids  =  8.6 kDa
Product: LacZα fragment of β-galactosidase
rrnB T1 terminator
285 .. 371  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
285 .. 371  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
558 .. 605  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
558 .. 605  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lacZα
2 .. 37  =  36 bp
12 amino acids  =  1.4 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2 .. 37  =  36 bp
12 amino acids  =  1.4 kDa
Product: LacZα fragment of β-galactosidase
tet promoter
87 .. 115  =  29 bp
3 segments
   Segment 3:  -10  
   87 .. 92  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
87 .. 115  =  29 bp
3 segments
   Segment 2:  
   93 .. 109  =  17 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
87 .. 115  =  29 bp
3 segments
   Segment 1:  -35  
   110 .. 115  =  6 bp
E. coli promoter for tetracycline efflux protein gene
tet promoter
87 .. 115  =  29 bp
3 segments
E. coli promoter for tetracycline efflux protein gene
rrnB T2 terminator
463 .. 490  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
463 .. 490  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
lac operator
3219 .. 3243  =  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
3219 .. 3243  =  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)
3152 .. 3171  =  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)
3152 .. 3171  =  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
3200 .. 3218  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3200 .. 3218  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
3464 .. 3469  =  6 bp
ribosome binding site
RBS
3464 .. 3469  =  6 bp
ribosome binding site
ORF:  1342 .. 1608  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3253 .. 3477  =  225 bp
ORF:  74 amino acids  =  8.6 kDa  (no start codon)
ORF:  144 .. 452  =  309 bp
ORF:  102 amino acids  =  11.4 kDa
ORF:  1212 .. 2072  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2913 .. 3272  =  360 bp
ORF:  119 amino acids  =  12.8 kDa
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Individual Sequences & Maps

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