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Plasmid Files

pBeloBAC11

Single-copy E. coli plasmid vector designed for constructing Bacterial Artificial Chromosomes (BACs).

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pBeloBAC11 Sequence and MappBeloBAC11.dna
Map and Sequence File   
Sequence Author:  New England Biolabs
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 HpaI (6997) BstEII (6972) SexAI * (6968) AhdI (6854) PciI (6850) ApaI (6340) PspOMI (6336) BsaI (6178) PfoI * (6119) SpeI (6090) AleI (5911) BclI * (5166) BseRI (5068) SnaBI (4999) PflFI - Tth111I (4639) BstXI (4453) MfeI (4355) NmeAIII (4307) AfeI (3934) ApaLI (87) KasI (145) BsaHI - NarI (146) SfoI (147) PluTI (149) BamHI (354) SphI (382) HindIII (384) lac operator SfiI - SrfI (640) ScaI (794) BstZ17I (1833) PaeR7I - PspXI - XhoI (2381) PsiI (2490) StuI (2542) EcoNI (2837) FspAI (3120) BsrGI (3148) BlpI (3847) pBeloBAC11 7507 bp
HpaI  (6997)
1 site
G T T A A C C A A T T G
BstEII  (6972)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
SexAI  (6968)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
AhdI  (6854)
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.
PciI  (6850)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaI  (6340)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (6336)
1 site
G G G C C C C C C G G G
BsaI  (6178)
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.
PfoI  (6119)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
SpeI  (6090)
1 site
A C T A G T T G A T C A
AleI  (5911)
1 site
C A C N N N N G T G G T G N N N N C A C
BclI  (5166)
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.
BseRI  (5068)
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.
SnaBI  (4999)
1 site
T A C G T A A T G C A T
PflFI  (4639)
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  (4639)
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.
BstXI  (4453)
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.
MfeI  (4355)
1 site
C A A T T G G T T A A C
NmeAIII  (4307)
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).
AfeI  (3934)
1 site
A G C G C T T C G C G A
ApaLI  (87)
1 site
G T G C A C C A C G T G
KasI  (145)
1 site
G G C G C C C C G C G G
BsaHI  (146)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
NarI  (146)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (147)
1 site
G G C G C C C C G C G G
PluTI  (149)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
BamHI  (354)
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.
SphI  (382)
1 site
G C A T G C C G T A C G
HindIII  (384)
1 site
A A G C T T T T C G A A
SfiI  (640)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
SrfI  (640)
1 site
G C C C G G G C C G G G C C C G
ScaI  (794)
1 site
A G T A C T T C A T G A
BstZ17I  (1833)
1 site
G T A T A C C A T A T G
PaeR7I  (2381)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2381)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2381)
1 site
C T C G A G G A G C T C
PsiI  (2490)
1 site
T T A T A A A A T A T T
StuI  (2542)
1 site
A G G C C T T C C G G A
EcoNI  (2837)
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.
FspAI  (3120)
1 site
R T G C G C A Y Y A C G C G T R
BsrGI  (3148)
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.
BlpI  (3847)
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.
sopA
4099 .. 5274  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F
plasmid
sopA
4099 .. 5274  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F
plasmid
repE
2765 .. 3520  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the
bacterial F plasmid
repE
2765 .. 3520  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the
bacterial F plasmid
CmR
766 .. 1425  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
766 .. 1425  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
sopC
6318 .. 6791  =  474 bp
centromere-like partitioning region of the bacterial F
plasmid
sopC
6318 .. 6791  =  474 bp
centromere-like partitioning region of the bacterial F
plasmid
cos
7051 .. 7449  =  399 bp
lambda cos site; allows packaging into phage
lambda particles
cos
7051 .. 7449  =  399 bp
lambda cos site; allows packaging into phage
lambda particles
lacZα
56 .. 436  =  381 bp
126 amino acids  =  14.5 kDa
Product: LacZα fragment of β-galactosidase
lacZα
56 .. 436  =  381 bp
126 amino acids  =  14.5 kDa
Product: LacZα fragment of β-galactosidase
incC
3523 .. 3773  =  251 bp
incompatibility region of the bacterial F plasmid
incC
3523 .. 3773  =  251 bp
incompatibility region of the bacterial F plasmid
ori2
2455 .. 2674  =  220 bp
secondary origin of replication for the bacterial F
plasmid; also known as oriS
ori2
2455 .. 2674  =  220 bp
secondary origin of replication for the bacterial F
plasmid; also known as oriS
cat promoter
1426 .. 1528  =  103 bp
promoter of the E. coli cat gene
cat promoter
1426 .. 1528  =  103 bp
promoter of the E. coli cat gene
loxP
7467 .. 7500  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
7467 .. 7500  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
lac promoter
480 .. 510  =  31 bp
   Segment 3:  -10  
   480 .. 486  =  7 bp
promoter for the E. coli lac operon
lac promoter
480 .. 510  =  31 bp
   Segment 2:  
   487 .. 504  =  18 bp
promoter for the E. coli lac operon
lac promoter
480 .. 510  =  31 bp
   Segment 1:  -35  
   505 .. 510  =  6 bp
promoter for the E. coli lac operon
lac promoter
480 .. 510  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
456 .. 472  =  17 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
456 .. 472  =  17 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).
sopB
5274 .. 6245  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F
plasmid
sopB
5274 .. 6245  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F
plasmid
MCS
333 .. 389  =  57 bp
pUC18/19 multiple cloning site
MCS
333 .. 389  =  57 bp
pUC18/19 multiple cloning site
T7 promoter
312 .. 330  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
312 .. 330  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
396 .. 414  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
396 .. 414  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 fwd
289 .. 305  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
289 .. 305  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
432 .. 448  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
432 .. 448  =  17 bp
common sequencing primer, one of multiple similar
variants
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