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Plasmid Files

pF1A T7

Flexi® vector with an ampicillin resistance marker, for bacterial or in vitro expression of an untagged protein.

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pF1A T7 Sequence and MappF1A T7.dna
Map and Sequence File   
Sequence Author:  Promega
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 T7 promoter BglII (3007) MluI (2888) BspEI (2675) DrdI (2019) PciI (1911) AgeI (1843) BstBI (1816) AhdI (1741) lac operator SpeI (67) AsiSI - PvuI - SgfI (104) BtgI - NcoI (107) DraI - PmeI (449) EcoRI (454) Eco53kI (462) BanII - SacI (464) Acc65I (466) AvaI - BsoBI - KpnI - TspMI - XmaI (470) BmeT110I (471) SmaI (472) BamHI (475) XbaI (481) SalI (487) AccI (488) HincII (489) PstI - SbfI (497) SphI (503) BlpI (562) EcoO109I (589) BsaAI - SnaBI (803) SspI (936) EarI (942) XmnI (1141) FspI (1518) BglI (1623) pF1A T7 3455 bp
BglII  (3007)
1 site
A G A T C T T C T A G A
MluI  (2888)
1 site
A C G C G T T G C G C A
BspEI  (2675)
1 site
T C C G G A A G G C C T
DrdI  (2019)
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  (1911)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (1843)
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  (1816)
1 site
T T C G A A A A G C T T
AhdI  (1741)
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.
SpeI  (67)
1 site
A C T A G T T G A T C A
AsiSI  (104)
1 site
G C G A T C G C C G C T A G C G
PvuI  (104)
1 site
C G A T C G G C T A G C
SgfI  (104)
1 site
G C G A T C G C C G C T A G C G
BtgI  (107)
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  (107)
1 site
C C A T G G G G T A C C
DraI  (449)
1 site
T T T A A A A A A T T T
PmeI  (449)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (454)
1 site
G A A T T C C T T A A G
Eco53kI  (462)
1 site
G A G C T C C T C G A G
BanII  (464)
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  (464)
1 site
G A G C T C C T C G A G
Acc65I  (466)
1 site
G G T A C C C C A T G G
AvaI  (470)
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  (470)
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.
KpnI  (470)
1 site
G G T A C C C C A T G G
TspMI  (470)
1 site
C C C G G G G G G C C C
XmaI  (470)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
BmeT110I  (471)
1 site
C Y C G R G G R G C Y C
SmaI  (472)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (475)
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  (481)
1 site
T C T A G A A G A T C T
SalI  (487)
1 site
G T C G A C C A G C T G
AccI  (488)
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  (489)
1 site
G T Y R A C C A R Y T G
PstI  (497)
1 site
C T G C A G G A C G T C
SbfI  (497)
1 site
C C T G C A G G G G A C G T C C
SphI  (503)
1 site
G C A T G C C G T A C G
BlpI  (562)
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  (589)
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  (803)
1 site
Y A C G T R R T G C A Y
SnaBI  (803)
1 site
T A C G T A A T G C A T
SspI  (936)
1 site
A A T A T T T T A T A A
EarI  (942)
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  (1141)
1 site
G A A N N N N T T C C T T N N N N A A G
FspI  (1518)
1 site
T G C G C A A C G C G T
BglI  (1623)
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.
AmpR
954 .. 1814  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   954 .. 1022  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
954 .. 1814  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1023 .. 1814  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
954 .. 1814  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1972 .. 2560  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1972 .. 2560  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
barnase
109 .. 444  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus
amyloliquefaciens

The barnase gene is lethal in standard bacterial
transformation strains.
barnase
109 .. 444  =  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
2676 .. 2959  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
cer region
2676 .. 2959  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
AmpR promoter
849 .. 953  =  105 bp
AmpR promoter
849 .. 953  =  105 bp
rrnB T1 terminator
3189 .. 3275  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
3189 .. 3275  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
T7 terminator
573 .. 620  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
573 .. 620  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
rrnB T2 terminator
3367 .. 3394  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
3367 .. 3394  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
lac operator
40 .. 64  =  25 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
40 .. 64  =  25 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).
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
21 .. 39  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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