pET-49b(+)

Bacterial vector for expressing proteins with a cleavable N-terminal GST tag and a cleavable C-terminal S-tag.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pET-49b(+)
PaeR7I - XhoI (147) PacI (141) AvrII (131) BlpI (80) DraIII (5684) PsiI (5556) NruI (4640) AcuI (4329) AlwNI (4197) BssSI - BssSαI (3954) BstZ17I (3552) PflFI - Tth111I (3526) S-Tag thrombin site Eco53kI (236) SacI (238) EagI - NotI (242) HindIII (249) SalI (255) PshAI (262) ZraI (264) AatII (266) AscI (270) StuI (280) BsrGI (283) EcoRI (289) BamHI (295) Acc65I (302) KpnI - TspMI - XmaI (306) SmaI (308) KflI (310) HRV 3C site SacII (338) 6xHis SpeI (417) ScaI (514) SwaI (660) MscI (880) NdeI (1087) XbaI (1125) lac operator SgrAI (1236) SphI (1392) BstAPI (1600) MluI (1917) BstEII (2098) NmeAIII (2123) PspOMI (2124) ApaI (2128) HpaI (2423) pET-49b(+) 5926 bp
PaeR7I  (147)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (147)
1 site		 C T C G A G G A G C T C
PacI  (141)
1 site		 T T A A T T A A A A T T A A T T
AvrII  (131)
1 site		 C C T A G G G G A T C C
BlpI  (80)
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  (5684)
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  (5556)
1 site		 T T A T A A A A T A T T
NruI  (4640)
1 site		 T C G C G A A G C G C T
AcuI  (4329)
1 site		 C T G A A G ( N ) 14 N N G A C T T C ( N ) 14
Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
AlwNI  (4197)
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.
BssSI  (3954)
1 site		 C A C G A G G T G C T C
BssSαI  (3954)
1 site		 C A C G A G G T G C T C
BstZ17I  (3552)
1 site		 G T A T A C C A T A T G
PflFI  (3526)
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  (3526)
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.
Eco53kI  (236)
1 site		 G A G C T C C T C G A G
SacI  (238)
1 site		 G A G C T C C T C G A G
EagI  (242)
1 site		 C G G C C G G C C G G C
NotI  (242)
1 site		 G C G G C C G C C G C C G G C G
HindIII  (249)
1 site		 A A G C T T T T C G A A
SalI  (255)
1 site		 G T C G A C C A G C T G
PshAI  (262)
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.
ZraI  (264)
1 site		 G A C G T C C T G C A G
AatII  (266)
1 site		 G A C G T C C T G C A G
AscI  (270)
1 site		 G G C G C G C C C C G C G C G G
StuI  (280)
1 site		 A G G C C T T C C G G A
BsrGI  (283)
1 site		 T G T A C A A C A T G T
BsrGI is typically used at 37°C, but is even more active at 60°C.
EcoRI  (289)
1 site		 G A A T T C C T T A A G
BamHI  (295)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
Acc65I  (302)
1 site		 G G T A C C C C A T G G
KpnI  (306)
1 site		 G G T A C C C C A T G G
TspMI  (306)
1 site		 C C C G G G G G G C C C
XmaI  (306)
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  (308)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
KflI  (310)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different KflI sites may not be compatible.
SacII  (338)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
SpeI  (417)
1 site		 A C T A G T T G A T C A
ScaI  (514)
1 site		 A G T A C T T C A T G A
SwaI  (660)
1 site		 A T T T A A A T T A A A T T T A
SwaI is typically used at 25°C, but is 50% active at 37°C.
MscI  (880)
1 site		 T G G C C A A C C G G T
NdeI  (1087)
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.
XbaI  (1125)
1 site		 T C T A G A A G A T C T
SgrAI  (1236)
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  (1392)
1 site		 G C A T G C C G T A C G
BstAPI  (1600)
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.
MluI  (1917)
1 site		 A C G C G T T G C G C A
BstEII  (2098)
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.
NmeAIII  (2123)
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).
PspOMI  (2124)
1 site		 G G G C C C C C C G G G
ApaI  (2128)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
HpaI  (2423)
1 site		 G T T A A C C A A T T G
lacI
1567 .. 2649  =  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
1567 .. 2649  =  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).
S-Tag
168 .. 212  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
168 .. 212  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
thrombin site
213 .. 230  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
213 .. 230  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
HRV 3C site
309 .. 332  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C protease
HRV 3C site
309 .. 332  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C protease
6xHis
342 .. 359  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
342 .. 359  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
GST
435 .. 1088  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
435 .. 1088  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
KanR
4552 .. 5367  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
4552 .. 5367  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
3842 .. 4430  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3842 .. 4430  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5460 .. 5915  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5460 .. 5915  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop
3221 .. 3412  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
3221 .. 3412  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
lacI promoter
1489 .. 1566  =  78 bp
lacI promoter
1489 .. 1566  =  78 bp
MCS
234 .. 308  =  75 bp
multiple cloning site
MCS
234 .. 308  =  75 bp
multiple cloning site
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
1133 .. 1157  =  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
1133 .. 1157  =  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
1158 .. 1176  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1158 .. 1176  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
1097 .. 1102  =  6 bp
ribosome binding site
RBS
1097 .. 1102  =  6 bp
ribosome binding site
ORF:  1690 .. 2649  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  4552 .. 5367  =  816 bp
ORF:  271 amino acids  =  31.0 kDa
ORF:  3188 .. 3412  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  1089 .. 1817  =  729 bp
ORF:  242 amino acids  =  25.5 kDa
ORF:  2406 .. 2669  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  137 .. 430  =  294 bp
ORF:  97 amino acids  =  11.0 kDa
ORF:  1265 .. 1528  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  2432 .. 2683  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  2821 .. 3189  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  141 .. 1088  =  948 bp
ORF:  315 amino acids  =  35.3 kDa
ORF:  1140 .. 1391  =  252 bp
ORF:  83 amino acids  =  8.6 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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