pET-43.1 Ek_LIC

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

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidspET & Duet Vectors (Novagen)pET-43.1 Ek_LIC
DraIII (7053) PsiI (6925) AhdI (6641) BglI (6523) FspI (6418) ScaI (6160) AlwNI (5447) PciI (5031) BspQI - SapI (4915) PflFI - Tth111I (4776) Bpu10I (4137) PpuMI (4037) HpaI (3673) AvrII (483) PacI (503) 6xHis PaeR7I - XhoI (530) HSV tag PmlI (576) EagI - NotI (592) HindIII (599) Acc65I (620) KpnI (624) SalI - SgrDI (626) PstI - SbfI (639) AscI (641) BsrGI (648) EcoRI (654) BamHI (660) Eco53kI (670) SacI (672) BseRI (699) PshAI (701) enterokinase site TspMI - XmaI (739) SmaI (741) thrombin site S-Tag SacII (817) 6xHis SpeI (848) BglII (963) BsmI (1007) MscI (1095) StuI (1148) BfuAI - BspMI (1297) NcoI (1423) NdeI (2337) RBS XbaI (2375) T7 promoter SphI (2642) EcoNI (2702) BstEII (3348) PspOMI (3374) ApaI (3378) pET-43.1 Ek/LIC 7295 bp
DraIII  (7053)
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  (6925)
1 site		 T T A T A A A A T A T T
AhdI  (6641)
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.
BglI  (6523)
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  (6418)
1 site		 T G C G C A A C G C G T
ScaI  (6160)
1 site		 A G T A C T T C A T G A
AlwNI  (5447)
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  (5031)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (4915)
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  (4915)
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.
PflFI  (4776)
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  (4776)
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.
Bpu10I  (4137)
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.
PpuMI  (4037)
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.
HpaI  (3673)
1 site		 G T T A A C C A A T T G
AvrII  (483)
1 site		 C C T A G G G G A T C C
PacI  (503)
1 site		 T T A A T T A A A A T T A A T T
PaeR7I  (530)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (530)
1 site		 C T C G A G G A G C T C
PmlI  (576)
1 site		 C A C G T G G T G C A C
EagI  (592)
1 site		 C G G C C G G C C G G C
NotI  (592)
1 site		 G C G G C C G C C G C C G G C G
HindIII  (599)
1 site		 A A G C T T T T C G A A
Acc65I  (620)
1 site		 G G T A C C C C A T G G
KpnI  (624)
1 site		 G G T A C C C C A T G G
SalI  (626)
1 site		 G T C G A C C A G C T G
SgrDI  (626)
1 site		 C G T C G A C G G C A G C T G C
PstI  (639)
1 site		 C T G C A G G A C G T C
SbfI  (639)
1 site		 C C T G C A G G G G A C G T C C
AscI  (641)
1 site		 G G C G C G C C C C G C G C G G
BsrGI  (648)
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  (654)
1 site		 G A A T T C C T T A A G
BamHI  (660)
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.
Eco53kI  (670)
1 site		 G A G C T C C T C G A G
SacI  (672)
1 site		 G A G C T C C T C G A G
BseRI  (699)
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  (701)
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.
TspMI  (739)
1 site		 C C C G G G G G G C C C
XmaI  (739)
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  (741)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
SacII  (817)
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  (848)
1 site		 A C T A G T T G A T C A
BglII  (963)
1 site		 A G A T C T T C T A G A
BsmI  (1007)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
MscI  (1095)
1 site		 T G G C C A A C C G G T
StuI  (1148)
1 site		 A G G C C T T C C G G A
BfuAI  (1297)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1297)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
NcoI  (1423)
1 site		 C C A T G G G G T A C C
NdeI  (2337)
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  (2375)
1 site		 T C T A G A A G A T C T
SphI  (2642)
1 site		 G C A T G C C G T A C G
EcoNI  (2702)
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.
BstEII  (3348)
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  (3374)
1 site		 G G G C C C C C C G G G
ApaI  (3378)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
6xHis
512 .. 529  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
512 .. 529  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
HSV tag
536 .. 568  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
536 .. 568  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
enterokinase site
701 .. 715  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
701 .. 715  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
thrombin site
734 .. 751  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
734 .. 751  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
S-Tag
767 .. 811  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
767 .. 811  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
6xHis
821 .. 838  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
821 .. 838  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
NusA
854 .. 2338  =  1485 bp
495 amino acids  =  54.9 kDa
Product: transcription elongation protein (N utilization substance protein A)
highly soluble in E. coli
NusA
854 .. 2338  =  1485 bp
495 amino acids  =  54.9 kDa
Product: transcription elongation protein (N utilization substance protein A)
highly soluble in E. coli
lacI
2817 .. 3899  =  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
2817 .. 3899  =  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
5854 .. 6714  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5854 .. 5922  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5854 .. 6714  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5923 .. 6714  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5854 .. 6714  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
5092 .. 5680  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5092 .. 5680  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6829 .. 7284  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6829 .. 7284  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop
4471 .. 4662  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
4471 .. 4662  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rrnB T1 terminator
260 .. 346  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
260 .. 346  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
lacI promoter
2739 .. 2816  =  78 bp
lacI promoter
2739 .. 2816  =  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
2383 .. 2407  =  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
2383 .. 2407  =  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
2408 .. 2426  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2408 .. 2426  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
2347 .. 2352  =  6 bp
ribosome binding site
RBS
2347 .. 2352  =  6 bp
ribosome binding site
MCS
530 .. 673  =  144 bp
multiple cloning site
MCS
530 .. 673  =  144 bp
multiple cloning site
ORF:  427 .. 852  =  426 bp
ORF:  141 amino acids  =  15.1 kDa
ORF:  4438 .. 4662  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  5854 .. 6714  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  179 .. 487  =  309 bp
ORF:  102 amino acids  =  11.4 kDa
ORF:  824 .. 2152  =  1329 bp
ORF:  442 amino acids  =  51.3 kDa
ORF:  2339 .. 3067  =  729 bp
ORF:  242 amino acids  =  25.5 kDa
ORF:  3656 .. 3919  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  948 .. 1184  =  237 bp
ORF:  78 amino acids  =  8.0 kDa
ORF:  2940 .. 3899  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  267 .. 509  =  243 bp
ORF:  80 amino acids  =  8.5 kDa
ORF:  4071 .. 4439  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  6318 .. 6584  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  509 .. 2338  =  1830 bp
ORF:  609 amino acids  =  67.1 kDa
ORF:  2390 .. 2641  =  252 bp
ORF:  83 amino acids  =  8.6 kDa
ORF:  2515 .. 2778  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  3682 .. 3933  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
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Download pET-43.1 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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