pIEx Bac-5 3C_LIC (linearized)

Linearized insect cell and baculovirus vector for ligation-independent cloning (LIC), encoding a cleavable N-terminal GST tag and a cleavable C-terminal Strep-Tag® II.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsInsect Cell VectorspIEx Bac-5 3C_LIC (linearized)
DraIII (4024) BanII (3950) NmeAIII (3020) BpmI (2942) AlwNI (2395) PspFI (2287) BseYI (2283) PmeI (1859) SgrAI (1571) AleI (1562) AgeI (1331) BmgBI (987) AatII (764) ZraI (762) SnaBI (483) BstXI (4631) NsiI (4648) BbvCI - Bpu10I (4880) BglII (5477) EcoRV (5750) NheI (6117) BmtI (6121) PvuII (6287) BsiWI (6295) EagI (6353) PfoI (6594) PpuMI (6598) BspEI (6711) BamHI (6745) BtgI - NcoI (6749) EcoNI (6761) BspQI - SapI (6830) MscI (6958) End (7449) Start (0) thrombin site AvrII (72) StuI (77) Bsu36I (79) IE1 terminator PflFI - Tth111I (332) BlpI (396) pIEx/Bac™-5 3C/LIC 7449 bp
DraIII  (4024)
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.
BanII  (3950)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
NmeAIII  (3020)
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).
BpmI  (2942)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AlwNI  (2395)
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.
PspFI  (2287)
1 site		 C C C A G C G G G T C G
BseYI  (2283)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PmeI  (1859)
1 site		 G T T T A A A C C A A A T T T G
SgrAI  (1571)
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.
AleI  (1562)
1 site		 C A C N N N N G T G G T G N N N N C A C
AgeI  (1331)
1 site		 A C C G G T T G G C C A
BmgBI  (987)
1 site		 C A C G T C G T G C A G
This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
AatII  (764)
1 site		 G A C G T C C T G C A G
ZraI  (762)
1 site		 G A C G T C C T G C A G
SnaBI  (483)
1 site		 T A C G T A A T G C A T
BstXI  (4631)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
NsiI  (4648)
1 site		 A T G C A T T A C G T A
BbvCI  (4880)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (4880)
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.
BglII  (5477)
1 site		 A G A T C T T C T A G A
EcoRV  (5750)
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.
NheI  (6117)
1 site		 G C T A G C C G A T C G
BmtI  (6121)
1 site		 G C T A G C C G A T C G
PvuII  (6287)
1 site		 C A G C T G G T C G A C
BsiWI  (6295)
1 site		 C G T A C G G C A T G C
BsiWI is typically used at 55°C, but is 50% active at 37°C.
EagI  (6353)
1 site		 C G G C C G G C C G G C
PfoI  (6594)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
PpuMI  (6598)
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.
BspEI  (6711)
1 site		 T C C G G A A G G C C T
BamHI  (6745)
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.
BtgI  (6749)
1 site		 C C R Y G G G G Y R C C
Sticky ends from different BtgI sites may not be compatible.
NcoI  (6749)
1 site		 C C A T G G G G T A C C
EcoNI  (6761)
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.
BspQI  (6830)
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  (6830)
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.
MscI  (6958)
1 site		 T G G C C A A C C G G T
End  (7449)
0 sites
Start  (0)
0 sites
AvrII  (72)
1 site		 C C T A G G G G A T C C
StuI  (77)
1 site		 A G G C C T T C C G G A
Bsu36I  (79)
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.
PflFI  (332)
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  (332)
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  (396)
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.
baculovirus recombination region (ORF1629)
420 .. 1777  =  1358 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
420 .. 1777  =  1358 bp
contains part of ORF1629
AmpR
2799 .. 3659  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2799 .. 3590  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2799 .. 3659  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3591 .. 3659  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2799 .. 3659  =  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)
4645 .. 5472  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
4645 .. 5472  =  828 bp
contains ORF603 and part of lef2
GST
6751 .. 7404  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
6751 .. 7404  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
HRV 3C site
7435 .. 7458  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C and PreScission proteases
HRV 3C site
7435 .. 7458  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C and PreScission proteases
IE1 promoter
5998 .. 6589  =  592 bp
promoter of the ie1 gene from the baculovirus Autographa californica
IE1 promoter
5998 .. 6589  =  592 bp
promoter of the ie1 gene from the baculovirus Autographa californica
ori
2040 .. 2628  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2040 .. 2628  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hr5 enhancer
5512 .. 5994  =  483 bp
baculovirus early transcription enhancer
hr5 enhancer
5512 .. 5994  =  483 bp
baculovirus early transcription enhancer
f1 ori
3791 .. 4246  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3791 .. 4246  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
IE1 terminator
85 .. 391  =  307 bp
terminator of the ie1 gene from the baculovirus Autographa californica
IE1 terminator
85 .. 391  =  307 bp
terminator of the ie1 gene from the baculovirus Autographa californica
p10 promoter
6599 .. 6708  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
6599 .. 6708  =  110 bp
baculovirus promoter for expression in insect cells
AmpR promoter
3660 .. 3764  =  105 bp
AmpR promoter
3660 .. 3764  =  105 bp
thrombin site
5 .. 22  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
5 .. 22  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
Strep-Tag II
29 .. 52  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
Strep-Tag II
29 .. 52  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
ORF1629
436 .. 1777  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
436 .. 1777  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF603
4807 .. 5412  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
4807 .. 5412  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
lef2
4645 .. 4769  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
4645 .. 4769  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
ORF:  2929 .. 3195  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6751 .. 7458  =  708 bp
ORF:  236 amino acids  =  27.5 kDa
ORF:  4389 .. 4769  =  381 bp
ORF:  126 amino acids  =  14.4 kDa
ORF:  2799 .. 3659  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4851 .. 5081  =  231 bp
ORF:  76 amino acids  =  9.2 kDa
ORF:  5963 .. 6529  =  567 bp
ORF:  188 amino acids  =  21.5 kDa
ORF:  436 .. 1689  =  1254 bp
ORF:  417 amino acids  =  46.0 kDa
ORF:  4807 .. 5412  =  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

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
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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