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Plasmid Files

pF25K ICE T7

Flexi® vector with a kanamycin resistance marker, for cell-free translation in an insect cell extract.

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pF25K ICE T7.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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AseI (20) AclI (3782) BsaI (3646) ScaI (3459) MluI (3375) BspEI (3162) BspHI (3118) ApaLI (2712) PciI (2398) AgeI (2330) BstBI (2303) RsrII (2137) NaeI (2123) AsiSI - PvuI - SgfI (90) BtgI - NcoI (93) PmeI (457) EcoRI (462) Acc65I (474) KpnI (478) BamHI (483) XbaI (489) SalI (495) AccI (496) HincII (497) SbfI (505) BlpI (570) AfeI (645) MfeI (676) SnaBI (687) AanI - PsiI (712) SwaI (777) EcoO109I (1034) NruI (1069) NotI (1074) StuI (1083) BstAPI (1319) XcmI (1437) BclI * (1462) MscI (1703) FspI (1723) PflFI - Tth111I (1739) BsrDI (1854) NgoMIV (2121) pF25K ICE T7 3942 bp
AseI  (20)
1 site
A T T A A T T A A T T A
AclI  (3782)
1 site
A A C G T T T T G C A A
BsaI  (3646)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
ScaI  (3459)
1 site
A G T A C T T C A T G A
MluI  (3375)
1 site
A C G C G T T G C G C A
BspEI  (3162)
1 site
T C C G G A A G G C C T
BspHI  (3118)
1 site
T C A T G A A G T A C T
ApaLI  (2712)
1 site
G T G C A C C A C G T G
PciI  (2398)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AgeI  (2330)
1 site
A C C G G T T G G C C A
BstBI  (2303)
1 site
T T C G A A A A G C T T
RsrII  (2137)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
NaeI  (2123)
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.
AsiSI  (90)
1 site
G C G A T C G C C G C T A G C G
PvuI  (90)
1 site
C G A T C G G C T A G C
SgfI  (90)
1 site
G C G A T C G C C G C T A G C G
BtgI  (93)
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  (93)
1 site
C C A T G G G G T A C C
PmeI  (457)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (462)
1 site
G A A T T C C T T A A G
Acc65I  (474)
1 site
G G T A C C C C A T G G
KpnI  (478)
1 site
G G T A C C C C A T G G
BamHI  (483)
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  (489)
1 site
T C T A G A A G A T C T
SalI  (495)
1 site
G T C G A C C A G C T G
AccI  (496)
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  (497)
1 site
G T Y R A C C A R Y T G
SbfI  (505)
1 site
C C T G C A G G G G A C G T C C
BlpI  (570)
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.
AfeI  (645)
1 site
A G C G C T T C G C G A
MfeI  (676)
1 site
C A A T T G G T T A A C
SnaBI  (687)
1 site
T A C G T A A T G C A T
AanI  (712)
1 site
T T A T A A A A T A T T
PsiI  (712)
1 site
T T A T A A A A T A T T
SwaI  (777)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
EcoO109I  (1034)
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.
NruI  (1069)
1 site
T C G C G A A G C G C T
NotI  (1074)
1 site
G C G G C C G C C G C C G G C G
StuI  (1083)
1 site
A G G C C T T C C G G A
BstAPI  (1319)
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.
XcmI  (1437)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BclI  (1462)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
MscI  (1703)
1 site
T G G C C A A C C G G T
FspI  (1723)
1 site
T G C G C A A C G C G T
PflFI  (1739)
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  (1739)
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.
BsrDI  (1854)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
NgoMIV  (2121)
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.
NeoR/KanR
1493 .. 2287  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
1493 .. 2287  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
2459 .. 3047  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2459 .. 3047  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
polyhedrin 3' UTR
611 .. 983  =  373 bp
3' UTR from the Autographa californica nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)
polyhedrin 3' UTR
611 .. 983  =  373 bp
3' UTR from the Autographa californica nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)
barnase
117 .. 452  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
117 .. 452  =  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
3163 .. 3446  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3163 .. 3446  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
rrnB T1 terminator
3676 .. 3762  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3676 .. 3762  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
1018 .. 1065  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1018 .. 1065  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
polyhedrin 5' UTR
40 .. 85  =  46 bp
5' UTR from the Malacosoma neustria nucleopolyhedrovirus polyhedrin gene (Ezure et al., 2006)
polyhedrin 5' UTR
40 .. 85  =  46 bp
5' UTR from the Malacosoma neustria nucleopolyhedrovirus polyhedrin gene (Ezure et al., 2006)
poly(A)
988 .. 1017  =  30 bp
synthetic poly(A) tract
poly(A)
988 .. 1017  =  30 bp
synthetic poly(A) tract
rrnB T2 terminator
3854 .. 3881  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
3854 .. 3881  =  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
ORF:  1493 .. 2287  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  117 .. 452  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  1665 .. 2051  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  3408 .. 3755  =  348 bp
ORF:  115 amino acids  =  12.4 kDa
ORF:  3517 .. 3843  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  1802 .. 2056  =  255 bp
ORF:  84 amino acids  =  9.8 kDa
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