pIEx-9 3C_LIC

Insect cell vector for ligation-independent cloning (LIC) to express proteins with an N-terminal Strep-Tag® II-HRV 3C cassette and a C-terminal 10xHis tag.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsInsect Cell VectorspIEx-9 3C_LIC
BspDI * - ClaI * (207) BspQI - SapI (3498) PciI (3381) DrdI (3279) PspFI (3081) BseYI (3077) AlwNI (2972) AhdI (2493) BsaI (2427) BsrFI (2408) NmeAIII (2346) ScaI (2012) XmnI (1893) AatII (1574) EcoRV (239) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BclI * (778) IE1 promoter BsiWI (784) MluI (872) BtgI - NcoI (1082) ATG BstBI (1109) HRV 3C site EcoNI (1135) EcoO109I - KflI - PpuMI (1141) Acc65I (1148) KpnI (1152) BamHI (1155) BsrGI (1161) SalI (1168) AccI (1169) NotI (1175) Eco53kI (1185) BanII - SacI (1187) thrombin site DraIII (1248) Bsu36I (1258) PacI (1287) IE1 terminator PflFI - Tth111I (1511) BtgZI (1534) XbaI (1566) ZraI (1572) pIEx™-9 3C/LIC 3747 bp
BspDI  (207)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (207)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
BspQI  (3498)
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  (3498)
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.
PciI  (3381)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
DrdI  (3279)
1 site		 G A C N N N N N N G T C C T G N N N N N N C A G
Sticky ends from different DrdI sites may not be compatible.
PspFI  (3081)
1 site		 C C C A G C G G G T C G
BseYI  (3077)
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.
AlwNI  (2972)
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.
AhdI  (2493)
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  (2427)
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.
BsrFI  (2408)
1 site		 R C C G G Y Y G G C C R
Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
NmeAIII  (2346)
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).
ScaI  (2012)
1 site		 A G T A C T T C A T G A
XmnI  (1893)
1 site		 G A A N N N N T T C C T T N N N N A A G
AatII  (1574)
1 site		 G A C G T C C T G C A G
EcoRV  (239)
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.
PsiI  (409)
1 site		 T T A T A A A A T A T T
NheI  (606)
1 site		 G C T A G C C G A T C G
BmtI  (610)
1 site		 G C T A G C C G A T C G
SphI  (614)
1 site		 G C A T G C C G T A C G
BsmI  (775)
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.
BclI  (778)
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.
BsiWI  (784)
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.
MluI  (872)
1 site		 A C G C G T T G C G C A
BtgI  (1082)
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  (1082)
1 site		 C C A T G G G G T A C C
BstBI  (1109)
1 site		 T T C G A A A A G C T T
EcoNI  (1135)
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.
EcoO109I  (1141)
1 site		 R G G N C C Y Y C C N G G R
Sticky ends from different EcoO109I sites may not be compatible.
KflI  (1141)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different KflI sites may not be compatible.
PpuMI  (1141)
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.
Acc65I  (1148)
1 site		 G G T A C C C C A T G G
KpnI  (1152)
1 site		 G G T A C C C C A T G G
BamHI  (1155)
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.
BsrGI  (1161)
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.
SalI  (1168)
1 site		 G T C G A C C A G C T G
AccI  (1169)
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.
NotI  (1175)
1 site		 G C G G C C G C C G C C G G C G
Eco53kI  (1185)
1 site		 G A G C T C C T C G A G
BanII  (1187)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
SacI  (1187)
1 site		 G A G C T C C T C G A G
DraIII  (1248)
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.
Bsu36I  (1258)
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.
PacI  (1287)
1 site		 T T A A T T A A A A T T A A T T
PflFI  (1511)
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  (1511)
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.
BtgZI  (1534)
1 site		 G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4
Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
XbaI  (1566)
1 site		 T C T A G A A G A T C T
ZraI  (1572)
1 site		 G A C G T C C T G C A G
AmpR
1706 .. 2566  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1706 .. 1774  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1706 .. 2566  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1775 .. 2566  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1706 .. 2566  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
IE1 promoter
487 .. 1078  =  592 bp
promoter of the ie1 gene from the baculovirus Autographa californica
IE1 promoter
487 .. 1078  =  592 bp
promoter of the ie1 gene from the baculovirus Autographa californica
ori
2737 .. 3325  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2737 .. 3325  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hr5 enhancer
1 .. 483  =  483 bp
baculovirus early transcription enhancer
hr5 enhancer
1 .. 483  =  483 bp
baculovirus early transcription enhancer
IE1 terminator
1264 .. 1571  =  308 bp
terminator of the ie1 gene from the baculovirus Autographa californica
IE1 terminator
1264 .. 1571  =  308 bp
terminator of the ie1 gene from the baculovirus Autographa californica
ATG
1084 .. 1086  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1084 .. 1086  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
Strep-Tag II
1093 .. 1116  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
Strep-Tag II
1093 .. 1116  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
HRV 3C site
1123 .. 1146  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C and PreScission proteases
HRV 3C site
1123 .. 1146  =  24 bp
8 amino acids  =  902.1 Da
Product: recognition and cleavage site for human rhinovirus 3C and PreScission proteases
thrombin site
1192 .. 1209  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
1192 .. 1209  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
10xHis
1216 .. 1245  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
10xHis
1216 .. 1245  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
AmpR promoter
1601 .. 1705  =  105 bp
AmpR promoter
1601 .. 1705  =  105 bp
ORF:  1706 .. 2566  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2170 .. 2436  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  452 .. 1018  =  567 bp
ORF:  188 amino acids  =  21.5 kDa
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Download pIEx-9 3C_LIC.dna file

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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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I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
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Coronavirus Resources
Insect Cell Vectors
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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
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Mammalian Expression Vectors
Structural Genomics Vectors

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