pTriEx-4 Ek_LIC (linearized)

Linearized insect, bacterial, and mammalian vector for ligation-independent cloning (LIC), encoding an N-terminal 6xHis-S-Tag and a C-terminal HSV-8xHis.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsInsect Cell VectorspTriEx-4 Ek_LIC (linearized)
AanI (1287) AanI (730) Bsu36I (196) AanI (3298) AanI (3387) CMV enhancer NcoI (5090) XcmI (5116) PflMI (5160) TspMI - XmaI (5192) SmaI (5194) End (5223) Start (0) EcoRV (20) Eco53kI (27) SacI (29) BamHI (33) EcoRI (39) BglII (47) AscI - BssHII (55) PstI - SbfI (65) Acc65I (73) AgeI (76) KpnI (77) BstBI (82) HindIII (85) EagI - NotI (92) PvuII (102) BstZ17I (107) PmlI (112) PaeR7I - XhoI (154) DraIII (186) pTriEx™-4 Ek/LIC 5223 bp
AanI  (1287)
4 sites		 T T A T A A A A T A T T
AanI  (730)
4 sites		 T T A T A A A A T A T T
Bsu36I  (196)
1 site		 C C T N A G G G G A N T C C
Sticky ends from different Bsu36I sites may not be compatible.
AanI  (3298)
4 sites		 T T A T A A A A T A T T
AanI  (3387)
4 sites		 T T A T A A A A T A T T
NcoI  (5090)
1 site		 C C A T G G G G T A C C
XcmI  (5116)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
PflMI  (5160)
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.
TspMI  (5192)
1 site		 C C C G G G G G G C C C
XmaI  (5192)
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  (5194)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
End  (5223)
0 sites
Start  (0)
0 sites
EcoRV  (20)
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.
Eco53kI  (27)
1 site		 G A G C T C C T C G A G
SacI  (29)
1 site		 G A G C T C C T C G A G
BamHI  (33)
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.
EcoRI  (39)
1 site		 G A A T T C C T T A A G
BglII  (47)
1 site		 A G A T C T T C T A G A
AscI  (55)
1 site		 G G C G C G C C C C G C G C G G
BssHII  (55)
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.
PstI  (65)
1 site		 C T G C A G G A C G T C
SbfI  (65)
1 site		 C C T G C A G G G G A C G T C C
Acc65I  (73)
1 site		 G G T A C C C C A T G G
AgeI  (76)
1 site		 A C C G G T T G G C C A
KpnI  (77)
1 site		 G G T A C C C C A T G G
BstBI  (82)
1 site		 T T C G A A A A G C T T
HindIII  (85)
1 site		 A A G C T T T T C G A A
EagI  (92)
1 site		 C G G C C G G C C G G C
NotI  (92)
1 site		 G C G G C C G C C G C C G G C G
PvuII  (102)
1 site		 C A G C T G G T C G A C
BstZ17I  (107)
1 site		 G T A T A C C A T A T G
PmlI  (112)
1 site		 C A C G T G G T G C A C
PaeR7I  (154)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (154)
1 site		 C T C G A G G A G C T C
DraIII  (186)
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.
AmpR
2056 .. 2916  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2056 .. 2847  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2056 .. 2916  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2848 .. 2916  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2056 .. 2916  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
baculovirus recombination region (lef2/ORF603)
3159 .. 3986  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
3159 .. 3986  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (ORF1629)
541 .. 1246  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
541 .. 1246  =  706 bp
contains part of ORF1629
Exon 1
4575 .. 4676  =  102 bp
Exon 1
4575 .. 4676  =  102 bp
intron
4677 .. 4840  =  164 bp
intron
4677 .. 4840  =  164 bp
Exon 2
4841 .. 5236  =  396 bp
Exon 2
4841 .. 5236  =  396 bp
ori
1456 .. 2043  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1456 .. 2043  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
Exon 2
1 .. 362  =  362 bp
Exon 2
1 .. 362  =  362 bp
CMV enhancer
4054 .. 4357  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4054 .. 4357  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
4359 .. 4558  =  200 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
4359 .. 4558  =  200 bp
human cytomegalovirus (CMV) immediate early promoter
T7 terminator
481 .. 528  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
481 .. 528  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
ORF603
3321 .. 3926  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
3321 .. 3926  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF1629
658 .. 1246  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
658 .. 1246  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ATG
5092 .. 5094  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
5092 .. 5094  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
5098 .. 5115  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
5098 .. 5115  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
S-Tag
5125 .. 5169  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
5125 .. 5169  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
thrombin site
5185 .. 5202  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
5185 .. 5202  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
enterokinase site
5221 .. 5235  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
5221 .. 5235  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
lef2
3159 .. 3283  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
3159 .. 3283  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
rabbit globin terminator
360 .. 471  =  112 bp
rabbit globin terminator
360 .. 471  =  112 bp
p10 promoter
4968 .. 5077  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
4968 .. 5077  =  110 bp
baculovirus promoter for expression in insect cells
HSV tag
121 .. 153  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
121 .. 153  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
8xHis
160 .. 183  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
160 .. 183  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
lac operator
4928 .. 4952  =  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
4928 .. 4952  =  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
4909 .. 4927  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4909 .. 4927  =  19 bp
promoter for bacteriophage T7 RNA polymerase
poly(A) signal
338 .. 343  =  6 bp
poly(A) signal
338 .. 343  =  6 bp
RBS
5079 .. 5084  =  6 bp
ribosome binding site
RBS
5079 .. 5084  =  6 bp
ribosome binding site
ORF:  2186 .. 2452  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2966 .. 3283  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  3365 .. 3595  =  231 bp
ORF:  76 amino acids  =  9.2 kDa
ORF:  4565 .. 4819  =  255 bp
ORF:  84 amino acids  =  9.0 kDa
ORF:  13 .. 324  =  312 bp
ORF:  103 amino acids  =  11.8 kDa
ORF:  658 .. 1275  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  2056 .. 2916  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3321 .. 3926  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

Plasmid Sets

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pET & Duet Vectors (Novagen)
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Coronavirus Resources
Insect Cell Vectors
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TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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