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Plasmid Files

pFusionRed-B

Vector for expressing the monomeric red fluorescent protein FusionRed in bacteria.

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pFusionRed-B Sequence and MappFusionRed-B.dna
Map and Sequence File   
Sequence Author:  Evrogen
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 AvaI - BsoBI - PaeR7I - XhoI (1) EcoO109I (4113) AatII (4059) ZraI (4057) XmnI (3736) PvuI (3507) FspI (3359) AseI (3309) AhdI (3137) AlwNI (2660) AflIII - PciI (2244) BspQI - SapI (2128) BstAPI (2068) NdeI (2067) BmeT110I (2) PsiI (49) MfeI (59) lac operator BseRI (114) ATG BsaBI * (125) BamHI (126) AgeI (134) AleI (147) BsrGI (187) Bsu36I (403) StuI (573) PshAI (777) DraIII (794) EagI - NotI (847) HindIII (854) BlpI (866) NheI (974) BmtI (978) Bpu10I (999) PvuII (1126) BspEI (1222) PasI (1458) XbaI (1831) PflFI - Tth111I (1991) BsaAI (1998) BstZ17I (2017) pFusionRed-B 4129 bp
AvaI  (1)
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  (1)
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  (1)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1)
1 site
C T C G A G G A G C T C
EcoO109I  (4113)
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.
AatII  (4059)
1 site
G A C G T C C T G C A G
ZraI  (4057)
1 site
G A C G T C C T G C A G
XmnI  (3736)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (3507)
1 site
C G A T C G G C T A G C
FspI  (3359)
1 site
T G C G C A A C G C G T
AseI  (3309)
1 site
A T T A A T T A A T T A
AhdI  (3137)
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.
AlwNI  (2660)
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.
AflIII  (2244)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2244)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2128)
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  (2128)
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.
BstAPI  (2068)
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.
NdeI  (2067)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
BmeT110I  (2)
1 site
C Y C G R G G R G C Y C
PsiI  (49)
1 site
T T A T A A A A T A T T
MfeI  (59)
1 site
C A A T T G G T T A A C
BseRI  (114)
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.
BsaBI  (125)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BamHI  (126)
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.
AgeI  (134)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AleI  (147)
1 site
C A C N N N N G T G G T G N N N N C A C
BsrGI  (187)
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.
Bsu36I  (403)
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.
StuI  (573)
1 site
A G G C C T T C C G G A
PshAI  (777)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
DraIII  (794)
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.
EagI  (847)
1 site
C G G C C G G C C G G C
NotI  (847)
1 site
G C G G C C G C C G C C G G C G
HindIII  (854)
1 site
A A G C T T T T C G A A
BlpI  (866)
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.
NheI  (974)
1 site
G C T A G C C G A T C G
BmtI  (978)
1 site
G C T A G C C G A T C G
Bpu10I  (999)
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.
PvuII  (1126)
1 site
C A G C T G G T C G A C
BspEI  (1222)
1 site
T C C G G A A G G C C T
PasI  (1458)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
XbaI  (1831)
1 site
T C T A G A A G A T C T
PflFI  (1991)
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  (1991)
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.
BsaAI  (1998)
1 site
Y A C G T R R T G C A Y
BstZ17I  (2017)
1 site
G T A T A C C A T A T G
AmpR
3064 .. 3924  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3064 .. 3855  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3064 .. 3924  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3856 .. 3924  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3064 .. 3924  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
FusionRed
147 .. 845  =  699 bp
232 amino acids  =  25.6 kDa
Product: monomeric red fluorescent protein
optimized for use as a fusion tag (Shemiakina et al.,
2012)
mammalian codon-optimized
FusionRed
147 .. 845  =  699 bp
232 amino acids  =  25.6 kDa
Product: monomeric red fluorescent protein
optimized for use as a fusion tag (Shemiakina et al.,
2012)
mammalian codon-optimized
CmR
1013 .. 1672  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1013 .. 1672  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
2305 .. 2893  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2305 .. 2893  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
3925 .. 4029  =  105 bp
AmpR promoter
3925 .. 4029  =  105 bp
lambda t0 terminator
875 .. 969  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
875 .. 969  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
1737 .. 1823  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
1737 .. 1823  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
T5 promoter
10 .. 54  =  45 bp
   Segment 1:  
   10 .. 24  =  15 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
   Segment 2:  -35  
   25 .. 30  =  6 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
   Segment 3:  
   31 .. 47  =  17 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
   Segment 4:  -10  
   48 .. 54  =  7 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
4 segments
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
lac operator
62 .. 78  =  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
62 .. 78  =  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).
ATG
114 .. 116  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
114 .. 116  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
lac operator
30 .. 46  =  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
30 .. 46  =  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).
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