pFC7A (HQ)

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

Sequence Author: Promega

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T7 promoter BglII (2977) MluI (2858) BspEI (2645) DrdI (1989) PciI (1881) AgeI (1813) BstBI (1786) BanI (1758) AsiSI - PvuI (65) BtgI - NcoI (68) AvaI - BsoBI - TspMI - XmaI (261) SmaI (263) Eco53kI (427) BanII - SacI (429) XbaI (451) SalI (457) AccI (458) HincII (459) PstI - SbfI (467) SphI (473) BlpI (532) EcoO109I (559) BsaAI - SnaBI (773) SspI (906) EarI (912) XmnI (1111) FspI (1488) BglI (1593) BpmI (1642) pFC7A (HQ) 3425 bp
BglII  (2977)
1 site
A G A T C T T C T A G A
MluI  (2858)
1 site
A C G C G T T G C G C A
BspEI  (2645)
1 site
T C C G G A A G G C C T
DrdI  (1989)
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  (1881)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (1813)
1 site
A C C G G T T G G C C A
BstBI  (1786)
1 site
T T C G A A A A G C T T
BanI  (1758)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
AsiSI  (65)
1 site
G C G A T C G C C G C T A G C G
PvuI  (65)
1 site
C G A T C G G C T A G C
BtgI  (68)
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  (68)
1 site
C C A T G G G G T A C C
AvaI  (261)
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  (261)
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.
TspMI  (261)
1 site
C C C G G G G G G C C C
XmaI  (261)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (263)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Eco53kI  (427)
1 site
G A G C T C C T C G A G
BanII  (429)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (429)
1 site
G A G C T C C T C G A G
XbaI  (451)
1 site
T C T A G A A G A T C T
SalI  (457)
1 site
G T C G A C C A G C T G
AccI  (458)
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  (459)
1 site
G T Y R A C C A R Y T G
PstI  (467)
1 site
C T G C A G G A C G T C
SbfI  (467)
1 site
C C T G C A G G G G A C G T C C
SphI  (473)
1 site
G C A T G C C G T A C G
BlpI  (532)
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  (559)
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  (773)
1 site
Y A C G T R R T G C A Y
SnaBI  (773)
1 site
T A C G T A A T G C A T
SspI  (906)
1 site
A A T A T T T T A T A A
EarI  (912)
1 site
C T C T T C N G A G A A G N N N N

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
XmnI  (1111)
1 site
G A A N N N N T T C C T T N N N N A A G
FspI  (1488)
1 site
T G C G C A A C G C G T
BglI  (1593)
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  (1642)
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
924 .. 1784  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   924 .. 992  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
924 .. 1784  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   993 .. 1784  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
924 .. 1784  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1942 .. 2530  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1942 .. 2530  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
70 .. 405  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
70 .. 405  =  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
2646 .. 2929  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
2646 .. 2929  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
AmpR promoter
819 .. 923  =  105 bp
AmpR promoter
819 .. 923  =  105 bp
rrnB T1 terminator
3159 .. 3245  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3159 .. 3245  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
543 .. 590  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
543 .. 590  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
rrnB T2 terminator
3337 .. 3364  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
3337 .. 3364  =  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
430 .. 447  =  18 bp
6 amino acids  =  813.8 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
430 .. 447  =  18 bp
6 amino acids  =  813.8 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).
ORF:  70 .. 405  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  3412 .. 358  =  372 bp
ORF:  123 amino acids  =  13.9 kDa
ORF:  2891 .. 3238  =  348 bp
ORF:  115 amino acids  =  12.4 kDa
ORF:  924 .. 1784  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3000 .. 3326  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  1388 .. 1654  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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