pMelBac C

Baculovirus transfer vector for expression and secretion of recombinant proteins. For other reading frames, use pMelBac A or pMelBac B.

Sequence Author: Thermo Fisher (Invitrogen)

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EcoRV (3) BstAPI (4732) Ac PCNA Bsu36I (4300) AleI (4235) Bpu10I (4019) BfuAI - BspMI (3749) PciI (3294) AlwNI (2885) BsaBI (28) polyhedrin promoter melittin signal sequence BstZ17I (137) BamHI (166) AvaI - BsoBI - PaeR7I - XhoI (175) BglII (179) PstI (187) Acc65I (191) KpnI - NcoI - StyI (195) EcoRI (200) BstBI (204) HindIII (207) SalI (216) BseRI (246) AfeI (355) BsaAI - SnaBI (397) PsiI (422) SwaI (487) baculovirus recombination region (ORF1629) BsrGI (727) BbsI (856) BmgBI (901) PfoI (1367) EcoO109I (1426) ScaI (1925) BsrFI (2321) BsaI (2340) AhdI (2406) pMelBac C 4829 bp
EcoRV  (3)
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.
BstAPI  (4732)
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.
Bsu36I  (4300)
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.
AleI  (4235)
1 site		 C A C N N N N G T G G T G N N N N C A C
Bpu10I  (4019)
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.
BfuAI  (3749)
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  (3749)
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.
PciI  (3294)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AlwNI  (2885)
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.
BsaBI  (28)
1 site		 G A T N N N N A T C C T A N N N N T A G
BstZ17I  (137)
1 site		 G T A T A C C A T A T G
BamHI  (166)
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.
AvaI  (175)
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  (175)
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  (175)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (175)
1 site		 C T C G A G G A G C T C
BglII  (179)
1 site		 A G A T C T T C T A G A
PstI  (187)
1 site		 C T G C A G G A C G T C
Acc65I  (191)
1 site		 G G T A C C C C A T G G
KpnI  (195)
1 site		 G G T A C C C C A T G G
NcoI  (195)
1 site		 C C A T G G G G T A C C
StyI  (195)
1 site		 C C W W G G G G W W C C
Sticky ends from different StyI sites may not be compatible.
EcoRI  (200)
1 site		 G A A T T C C T T A A G
BstBI  (204)
1 site		 T T C G A A A A G C T T
HindIII  (207)
1 site		 A A G C T T T T C G A A
SalI  (216)
1 site		 G T C G A C C A G C T G
BseRI  (246)
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.
AfeI  (355)
1 site		 A G C G C T T C G C G A
BsaAI  (397)
1 site		 Y A C G T R R T G C A Y
SnaBI  (397)
1 site		 T A C G T A A T G C A T
PsiI  (422)
1 site		 T T A T A A A A T A T T
SwaI  (487)
1 site		 A T T T A A A T T A A A T T T A
SwaI is typically used at 25°C, but is 50% active at 37°C.
BsrGI  (727)
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.
BbsI  (856)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BmgBI  (901)
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.
PfoI  (1367)
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.
EcoO109I  (1426)
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.
ScaI  (1925)
1 site		 A G T A C T T C A T G A
BsrFI  (2321)
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.
BsaI  (2340)
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.
AhdI  (2406)
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.
5' lacZ fragment
3415 .. 4517  =  1103 bp
367 amino acids  =  41.8 kDa
Product: N-terminal fragment of β-galactosidase
5' lacZ fragment
3415 .. 4517  =  1103 bp
367 amino acids  =  41.8 kDa
Product: N-terminal fragment of β-galactosidase
Ac PCNA
4525 .. 4614  =  90 bp
30 amino acids  =  3.6 kDa
Product: proliferating cell nuclear antigen (PCNA) from baculovirus
Ac PCNA
4525 .. 4614  =  90 bp
30 amino acids  =  3.6 kDa
Product: proliferating cell nuclear antigen (PCNA) from baculovirus
baculovirus recombination region (ORF1629)
233 .. 1177  =  945 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
233 .. 1177  =  945 bp
contains part of ORF1629
AmpR
1619 .. 2479  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1619 .. 1687  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1619 .. 2479  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1688 .. 2479  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1619 .. 2479  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2650 .. 3238  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2650 .. 3238  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ETL promoter
4615 .. 4829  =  215 bp
baculovirus early-to-late promoter (Liu et al., 2006)
ETL promoter
4615 .. 4829  =  215 bp
baculovirus early-to-late promoter (Liu et al., 2006)
AmpR promoter
1514 .. 1618  =  105 bp
AmpR promoter
1514 .. 1618  =  105 bp
polyhedrin promoter
4 .. 95  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
4 .. 95  =  92 bp
promoter for the baculovirus polyhedrin gene
melittin signal sequence
96 .. 158  =  63 bp
21 amino acids  =  2.5 kDa
Product: signal sequence from honeybee melittin
melittin signal sequence
96 .. 158  =  63 bp
21 amino acids  =  2.5 kDa
Product: signal sequence from honeybee melittin
MCS
166 .. 221  =  56 bp
multiple cloning site
MCS
166 .. 221  =  56 bp
multiple cloning site
ORF:  1619 .. 2479  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4613 .. 20  =  237 bp
ORF:  78 amino acids
ORF:  96 .. 320  =  225 bp
ORF:  74 amino acids  =  8.6 kDa
ORF:  4637 .. 32  =  225 bp
ORF:  74 amino acids  =  8.4 kDa
ORF:  105 .. 332  =  228 bp
ORF:  75 amino acids  =  9.2 kDa
ORF:  350 .. 1096  =  747 bp
ORF:  248 amino acids  =  27.9 kDa
ORF:  2083 .. 2349  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3292 .. 4614  =  1323 bp
ORF:  440 amino acids  =  50.0 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

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