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Plasmid Files

pFN6A (HQ)

Flexi® vector with an ampicillin resistance marker, for bacterial or in vitro expression of a protein with an N-terminal HQ metal affinity tag.

 
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 T7 promoter BglII (3023) MluI (2904) BspEI (2691) DrdI (2035) PciI (1927) AgeI (1859) BstBI (1832) AhdI (1757) ATG HQ tag AsiSI - PvuI - SgfI (98) BtgI - NcoI (101) DraI - PmeI (465) BanII (480) Acc65I (482) KpnI (486) BamHI (491) XbaI (497) SalI (503) AccI (504) HincII (505) PstI - SbfI (513) SphI (519) BlpI (578) EcoO109I (605) BsaAI - SnaBI (819) SspI (952) EarI (958) XmnI (1157) FspI (1534) BglI (1639) BpmI (1688) pFN6A (HQ) 3471 bp
BglII  (3023)
1 site
A G A T C T T C T A G A
MluI  (2904)
1 site
A C G C G T T G C G C A
BspEI  (2691)
1 site
T C C G G A A G G C C T
DrdI  (2035)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PciI  (1927)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (1859)
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.
BstBI  (1832)
1 site
T T C G A A A A G C T T
AhdI  (1757)
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.
AsiSI  (98)
1 site
G C G A T C G C C G C T A G C G
PvuI  (98)
1 site
C G A T C G G C T A G C
SgfI  (98)
1 site
G C G A T C G C C G C T A G C G
BtgI  (101)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (101)
1 site
C C A T G G G G T A C C
DraI  (465)
1 site
T T T A A A A A A T T T
PmeI  (465)
1 site
G T T T A A A C C A A A T T T G
BanII  (480)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
Acc65I  (482)
1 site
G G T A C C C C A T G G
KpnI  (486)
1 site
G G T A C C C C A T G G
BamHI  (491)
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.
XbaI  (497)
1 site
T C T A G A A G A T C T
SalI  (503)
1 site
G T C G A C C A G C T G
AccI  (504)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
HincII  (505)
1 site
G T Y R A C C A R Y T G
PstI  (513)
1 site
C T G C A G G A C G T C
SbfI  (513)
1 site
C C T G C A G G G G A C G T C C
SphI  (519)
1 site
G C A T G C C G T A C G
BlpI  (578)
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  (605)
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.
BsaAI  (819)
1 site
Y A C G T R R T G C A Y
SnaBI  (819)
1 site
T A C G T A A T G C A T
SspI  (952)
1 site
A A T A T T T T A T A A
EarI  (958)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI
recognition sequence.
Sticky ends from different EarI sites may not be compatible.
XmnI  (1157)
1 site
G A A N N N N T T C C T T N N N N A A G
FspI  (1534)
1 site
T G C G C A A C G C G T
BglI  (1639)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
BpmI  (1688)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AmpR
970 .. 1830  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   970 .. 1038  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
970 .. 1830  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1039 .. 1830  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
970 .. 1830  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1988 .. 2576  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1988 .. 2576  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
barnase
125 .. 460  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus
amyloliquefaciens

The barnase gene is lethal in standard bacterial
transformation strains.
barnase
125 .. 460  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus
amyloliquefaciens

The barnase gene is lethal in standard bacterial
transformation strains.
cer region
2692 .. 2975  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
cer region
2692 .. 2975  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
AmpR promoter
865 .. 969  =  105 bp
AmpR promoter
865 .. 969  =  105 bp
rrnB T1 terminator
3205 .. 3291  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
3205 .. 3291  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
T7 terminator
589 .. 636  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
589 .. 636  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
rrnB T2 terminator
3383 .. 3410  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
3383 .. 3410  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
HQ tag
76 .. 93  =  18 bp
6 amino acids  =  813.9 Da
Product: HQHQHQ metal affinity tag
The HQ tag binds to nickel and other metal ions
much like the 6xHis tag, but may allow elution at
lower concentrations of imidazole (Godat et al.,
2008).
HQ tag
76 .. 93  =  18 bp
6 amino acids  =  813.9 Da
Product: HQHQHQ metal affinity tag
The HQ tag binds to nickel and other metal ions
much like the 6xHis tag, but may allow elution at
lower concentrations of imidazole (Godat et al.,
2008).
ATG
70 .. 72  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
70 .. 72  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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