pET-31b(+)

Bacterial vector for expressing ketosteroid isomerase fusion proteins.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pET-31b(+)
StyI (57) DraIII (5500) PsiI (5372) ScaI (4837) PvuI (4727) PstI (4602) BsaI (4418) AhdI (4357) PciI (3470) BspQI - SapI (3354) BstZ17I (3241) AccI (3240) PflFI - Tth111I (3215) BlpI (80) AvaI - BsoBI - PaeR7I - PspXI - XhoI (158) AlwNI (169) BspDI * - ClaI * (249) AleI (519) NdeI - NsiI (543) XbaI (581) T7 promoter BglII (647) SgrAI (688) EcoNI (904) PflMI (951) BstAPI (1052) MluI (1369) BclI * (1383) BstEII (1550) PspOMI (1576) ApaI (1580) BssHII (1780) EcoRV (1819) HincII - HpaI (1875) PshAI (2214) FspAI (2451) PpuMI (2476) Bpu10I (2576) pET-31b(+) 5742 bp
StyI  (57)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
DraIII  (5500)
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  (5372)
1 site		 T T A T A A A A T A T T
ScaI  (4837)
1 site		 A G T A C T T C A T G A
PvuI  (4727)
1 site		 C G A T C G G C T A G C
PstI  (4602)
1 site		 C T G C A G G A C G T C
BsaI  (4418)
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.
AhdI  (4357)
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.
PciI  (3470)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (3354)
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  (3354)
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.
BstZ17I  (3241)
1 site		 G T A T A C C A T A T G
AccI  (3240)
1 site		 G T M K A C C A K M T G
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
PflFI  (3215)
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  (3215)
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.
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.
AvaI  (158)
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  (158)
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.
PaeR7I  (158)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (158)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (158)
1 site		 C T C G A G G A G C T C
AlwNI  (169)
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.
BspDI  (249)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (249)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
AleI  (519)
1 site		 C A C N N N N G T G G T G N N N N C A C
NdeI  (543)
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  (543)
1 site		 A T G C A T T A C G T A
XbaI  (581)
1 site		 T C T A G A A G A T C T
BglII  (647)
1 site		 A G A T C T T C T A G A
SgrAI  (688)
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.
EcoNI  (904)
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.
PflMI  (951)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
BstAPI  (1052)
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  (1369)
1 site		 A C G C G T T G C G C A
BclI  (1383)
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  (1550)
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.
PspOMI  (1576)
1 site		 G G G C C C C C C G G G
ApaI  (1580)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BssHII  (1780)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
EcoRV  (1819)
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.
HincII  (1875)
1 site		 G T Y R A C C A R Y T G
HpaI  (1875)
1 site		 G T T A A C C A A T T G
PshAI  (2214)
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.
FspAI  (2451)
1 site		 R T G C G C A Y Y A C G C G T R
PpuMI  (2476)
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.
Bpu10I  (2576)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
lacI
1019 .. 2101  =  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
1019 .. 2101  =  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
4284 .. 5144  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   4284 .. 5075  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4284 .. 5144  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   5076 .. 5144  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4284 .. 5144  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3531 .. 4113  =  583 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3531 .. 4113  =  583 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5276 .. 5731  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5276 .. 5731  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
KSI
170 .. 544  =  375 bp
125 amino acids  =  13.5 kDa
Product: ketosteroid isomerase
KSI
170 .. 544  =  375 bp
125 amino acids  =  13.5 kDa
Product: ketosteroid isomerase
rop
2910 .. 3101  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
2910 .. 3101  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
AmpR promoter
5145 .. 5249  =  105 bp
AmpR promoter
5145 .. 5249  =  105 bp
lacI promoter
941 .. 1018  =  78 bp
lacI promoter
941 .. 1018  =  78 bp
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
589 .. 613  =  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
589 .. 613  =  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
614 .. 632  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
614 .. 632  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
553 .. 558  =  6 bp
ribosome binding site
RBS
553 .. 558  =  6 bp
ribosome binding site
ORF:  769 .. 1269  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
ORF:  1858 .. 2121  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  4414 .. 4680  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  539 .. 772  =  234 bp
ORF:  77 amino acids  =  8.2 kDa
ORF:  1142 .. 2101  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  2157 .. 2513  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  2877 .. 3101  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  604 .. 843  =  240 bp
ORF:  79 amino acids  =  8.0 kDa
ORF:  717 .. 980  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  1884 .. 2135  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  4284 .. 5144  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  137 .. 544  =  408 bp
ORF:  135 amino acids  =  14.8 kDa
ORF:  2510 .. 2878  =  369 bp
ORF:  122 amino acids  =  14.2 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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