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pMelBac A

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

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pMelBac A.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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BstAPI (4734) Bsu36I (4302) AleI (4237) Bpu10I (4021) BfuAI - BspMI (3751) PciI (3296) AlwNI (2887) EcoRV (3) BstZ17I (137) BamHI (168) Eco53kI (176) AvaI - BsoBI - PaeR7I - XhoI (177) BanII - BmeT110I - SacI (178) BglII (181) PstI (189) Acc65I (193) KpnI - NcoI - StyI (197) EcoRI (202) BstBI (206) HindIII (209) SalI (218) BseRI (248) AfeI (357) BsaAI - SnaBI (399) PsiI (424) SwaI (489) BsrGI (729) BbsI (858) BmgBI (903) PfoI (1369) EcoO109I (1428) ScaI (1927) BsrFI (2323) BsaI (2342) AhdI (2408) pMelBac A 4831 bp
BstAPI  (4734)
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  (4302)
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  (4237)
1 site
C A C N N N N G T G G T G N N N N C A C
Bpu10I  (4021)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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  (3751)
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  (3751)
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  (3296)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (2887)
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.
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.
BstZ17I  (137)
1 site
G T A T A C C A T A T G
BamHI  (168)
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  (176)
1 site
G A G C T C C T C G A G
AvaI  (177)
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  (177)
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  (177)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (177)
1 site
C T C G A G G A G C T C
BanII  (178)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BmeT110I  (178)
1 site
C Y C G R G G R G C Y C
SacI  (178)
1 site
G A G C T C C T C G A G
BglII  (181)
1 site
A G A T C T T C T A G A
PstI  (189)
1 site
C T G C A G G A C G T C
Acc65I  (193)
1 site
G G T A C C C C A T G G
KpnI  (197)
1 site
G G T A C C C C A T G G
NcoI  (197)
1 site
C C A T G G G G T A C C
StyI  (197)
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  (202)
1 site
G A A T T C C T T A A G
BstBI  (206)
1 site
T T C G A A A A G C T T
HindIII  (209)
1 site
A A G C T T T T C G A A
SalI  (218)
1 site
G T C G A C C A G C T G
BseRI  (248)
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  (357)
1 site
A G C G C T T C G C G A
BsaAI  (399)
1 site
Y A C G T R R T G C A Y
SnaBI  (399)
1 site
T A C G T A A T G C A T
PsiI  (424)
1 site
T T A T A A A A T A T T
SwaI  (489)
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  (729)
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  (858)
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  (903)
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  (1369)
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  (1428)
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  (1927)
1 site
A G T A C T T C A T G A
BsrFI  (2323)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
BsaI  (2342)
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  (2408)
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
3417 .. 4519  =  1103 bp
367 amino acids  =  41.8 kDa
Product: N-terminal fragment of β-galactosidase
5' lacZ fragment
3417 .. 4519  =  1103 bp
367 amino acids  =  41.8 kDa
Product: N-terminal fragment of β-galactosidase
baculovirus recombination region (ORF1629)
235 .. 1179  =  945 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
235 .. 1179  =  945 bp
contains part of ORF1629
AmpR
1621 .. 2481  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1621 .. 1689  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1621 .. 2481  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1690 .. 2481  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1621 .. 2481  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2652 .. 3240  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2652 .. 3240  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ETL promoter
4617 .. 4831  =  215 bp
baculovirus early-to-late promoter (Liu et al., 2006)
ETL promoter
4617 .. 4831  =  215 bp
baculovirus early-to-late promoter (Liu et al., 2006)
AmpR promoter
1516 .. 1620  =  105 bp
AmpR promoter
1516 .. 1620  =  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
Ac PCNA
4527 .. 4616  =  90 bp
30 amino acids  =  3.6 kDa
Product: proliferating cell nuclear antigen (PCNA) from baculovirus
Ac PCNA
4527 .. 4616  =  90 bp
30 amino acids  =  3.6 kDa
Product: proliferating cell nuclear antigen (PCNA) from baculovirus
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
168 .. 223  =  56 bp
multiple cloning site
MCS
168 .. 223  =  56 bp
multiple cloning site
ORF1629
352 .. 1179  =  828 bp
275 amino acids  =  30.5 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
352 .. 1179  =  828 bp
275 amino acids  =  30.5 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF:  1621 .. 2481  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4615 .. 20  =  237 bp
ORF:  78 amino acids
ORF:  352 .. 1098  =  747 bp
ORF:  248 amino acids  =  27.9 kDa
ORF:  4639 .. 32  =  225 bp
ORF:  74 amino acids  =  8.4 kDa
ORF:  2085 .. 2351  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3294 .. 4616  =  1323 bp
ORF:  440 amino acids  =  50.0 kDa
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