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Plasmid Files

pRSET-BFP

Vector for expressing BFP in bacteria.

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pRSET-BFP Sequence and MappRSET-BFP.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 BspQI - SapI (3526) NspI (3413) AflIII - PciI (3409) PspFI (3109) BseYI (3105) AlwNI (3000) AhdI (2521) ScaI (2040) XmnI (1921) XbaI (47) NdeI (87) 6xHis NheI (125) BmtI (129) enterokinase site BspDI * - ClaI * (184) BamHI (191) EcoRI (197) NcoI (207) BseRI (239) PfoI * (489) BsrGI (918) AvaI - BsoBI - PaeR7I - XhoI (929) BmeT110I (930) HindIII (935) BlpI (993) EcoO109I (1020) BspEI * (1070) NgoMIV (1273) NaeI (1275) BanII (1307) BsaAI (1378) DraIII (1381) PsiI (1506) pRSET-BFP 3600 bp
BspQI  (3526)
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  (3526)
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.
NspI  (3413)
1 site
R C A T G Y Y G T A C R
AflIII  (3409)
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  (3409)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (3109)
1 site
C C C A G C G G G T C G
BseYI  (3105)
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  (3000)
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  (2521)
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.
ScaI  (2040)
1 site
A G T A C T T C A T G A
XmnI  (1921)
1 site
G A A N N N N T T C C T T N N N N A A G
XbaI  (47)
1 site
T C T A G A A G A T C T
NdeI  (87)
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.
NheI  (125)
1 site
G C T A G C C G A T C G
BmtI  (129)
1 site
G C T A G C C G A T C G
BspDI  (184)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (184)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BamHI  (191)
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.
EcoRI  (197)
1 site
G A A T T C C T T A A G
NcoI  (207)
1 site
C C A T G G G G T A C C
BseRI  (239)
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.
PfoI  (489)
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.
BsrGI  (918)
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.
AvaI  (929)
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  (929)
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  (929)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (929)
1 site
C T C G A G G A G C T C
BmeT110I  (930)
1 site
C Y C G R G G R G C Y C
HindIII  (935)
1 site
A A G C T T T T C G A A
BlpI  (993)
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.
EcoO109I  (1020)
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.
BspEI  (1070)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
NgoMIV  (1273)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (1275)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
BanII  (1307)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BsaAI  (1378)
1 site
Y A C G T R R T G C A Y
DraIII  (1381)
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.
PsiI  (1506)
1 site
T T A T A A A A T A T T
AmpR
1734 .. 2594  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1734 .. 1802  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1734 .. 2594  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1803 .. 2594  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1734 .. 2594  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
BFP
209 .. 928  =  720 bp
239 amino acids  =  26.8 kDa
   Segment 1:  
   209 .. 211  =  3 bp
   1 amino acid  =  149.2 Da
Product: blue variant of GFP
mammalian codon-optimized
BFP
209 .. 928  =  720 bp
239 amino acids  =  26.8 kDa
   Segment 2:  1a  
   212 .. 214  =  3 bp
   1 amino acid  =  117.2 Da
Product: blue variant of GFP
mammalian codon-optimized
BFP
209 .. 928  =  720 bp
239 amino acids  =  26.8 kDa
   Segment 3:  
   215 .. 928  =  714 bp
   237 amino acids  =  26.6 kDa
Product: blue variant of GFP
mammalian codon-optimized
BFP
209 .. 928  =  720 bp
239 amino acids  =  26.8 kDa
3 segments
Product: blue variant of GFP
mammalian codon-optimized
ori
2765 .. 3353  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2765 .. 3353  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1148 .. 1603  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1148 .. 1603  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
AmpR promoter
1629 .. 1733  =  105 bp
AmpR promoter
1629 .. 1733  =  105 bp
T7 terminator
1004 .. 1051  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
1004 .. 1051  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 tag
122 .. 154  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 tag
122 .. 154  =  33 bp
11 amino acids  =  1.1 kDa
Product: epitope tag from a T7 major capsid protein
T7 promoter
9 .. 27  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
9 .. 27  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
101 .. 118  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
101 .. 118  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
enterokinase site
167 .. 181  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
167 .. 181  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
ATG
89 .. 91  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
89 .. 91  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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