pET-46 Ek_LIC

Bacterial vector for ligation-independent cloning (LIC) to express 6xHis-tagged proteins with an enterokinase site.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pET-46 Ek_LIC
DraIII (4958) PsiI (4830) SspI (4750) AhdI (4546) BsaI (4480) BglI (4428) FspI (4323) PvuI (4177) ScaI (4065) AlwNI (3352) PciI (2936) BspQI - SapI (2820) BstZ17I (2707) PflFI - Tth111I (2681) BlpI (80) AvrII (97) PacI (107) AvaI - BsoBI - PaeR7I - XhoI (176) BseRI (201) PshAI (203) enterokinase site PmlI (220) NcoI (241) ATG RBS XbaI (280) lac operator T7 promoter BspDI * - ClaI * (349) SgrAI (391) SphI (547) EcoNI (607) BstAPI (755) MluI (1072) BclI * (1086) BstEII (1253) PspOMI (1279) ApaI (1283) BssHII (1483) HincII - HpaI (1578) PpuMI (1942) Bpu10I (2042) pET-46 Ek/LIC 5200 bp
DraIII  (4958)
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  (4830)
1 site		 T T A T A A A A T A T T
SspI  (4750)
1 site		 A A T A T T T T A T A A
AhdI  (4546)
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  (4480)
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  (4428)
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  (4323)
1 site		 T G C G C A A C G C G T
PvuI  (4177)
1 site		 C G A T C G G C T A G C
ScaI  (4065)
1 site		 A G T A C T T C A T G A
AlwNI  (3352)
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.
PciI  (2936)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (2820)
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  (2820)
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  (2707)
1 site		 G T A T A C C A T A T G
PflFI  (2681)
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  (2681)
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.
AvrII  (97)
1 site		 C C T A G G G G A T C C
PacI  (107)
1 site		 T T A A T T A A A A T T A A T T
AvaI  (176)
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  (176)
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  (176)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (176)
1 site		 C T C G A G G A G C T C
BseRI  (201)
1 site		 G A G G A G ( N ) 8 N N C T C C T C ( N ) 8
Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
PshAI  (203)
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.
PmlI  (220)
1 site		 C A C G T G G T G C A C
NcoI  (241)
1 site		 C C A T G G G G T A C C
XbaI  (280)
1 site		 T C T A G A A G A T C T
BspDI  (349)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (349)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
SgrAI  (391)
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  (547)
1 site		 G C A T G C C G T A C G
EcoNI  (607)
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  (755)
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  (1072)
1 site		 A C G C G T T G C G C A
BclI  (1086)
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  (1253)
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  (1279)
1 site		 G G G C C C C C C G G G
ApaI  (1283)
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  (1483)
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.
HincII  (1578)
1 site		 G T Y R A C C A R Y T G
HpaI  (1578)
1 site		 G T T A A C C A A T T G
PpuMI  (1942)
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  (2042)
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
722 .. 1804  =  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
722 .. 1804  =  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
3759 .. 4619  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3759 .. 3827  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3759 .. 4619  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3828 .. 4619  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3759 .. 4619  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2997 .. 3585  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2997 .. 3585  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
4734 .. 5189  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
4734 .. 5189  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop
2376 .. 2567  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
2376 .. 2567  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
S-Tag
125 .. 169  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
125 .. 169  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
enterokinase site
203 .. 217  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
203 .. 217  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
6xHis
221 .. 238  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
221 .. 238  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
242 .. 244  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
242 .. 244  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
lacI promoter
644 .. 721  =  78 bp
lacI promoter
644 .. 721  =  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
288 .. 312  =  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
288 .. 312  =  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
313 .. 331  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
313 .. 331  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
252 .. 257  =  6 bp
ribosome binding site
RBS
252 .. 257  =  6 bp
ribosome binding site
ORF:  472 .. 972  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
ORF:  1561 .. 1824  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  845 .. 1804  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  2343 .. 2567  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  3759 .. 4619  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1976 .. 2344  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  4223 .. 4489  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  295 .. 546  =  252 bp
ORF:  83 amino acids  =  8.6 kDa
ORF:  420 .. 683  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  1587 .. 1838  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
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Download pET-46 Ek_LIC.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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