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Plasmid Files

pFN2K (GST)

Flexi® vector with a kanamycin resistance marker, for bacterial or in vitro expression of a protein with a cleavable N-terminal GST tag.

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pFN2K (GST).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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AseI (20) AclI (3972) BsaI (3836) MluI (3565) BspHI (3308) ApaLI (2902) AgeI (2520) RsrII (2327) NaeI (2313) NgoMIV (2311) EcoNI (80) BsgI (356) XmnI (463) AsiSI - PvuI - SgfI (764) BtgI - NcoI (767) ZraI (925) AatII (927) BpmI (965) DraI - PmeI (1131) EcoRI (1136) Eco53kI (1144) SacI (1146) Acc65I (1148) AvaI - BsoBI - KpnI - TspMI - XmaI (1152) BmeT110I (1153) SmaI (1154) BamHI (1157) XbaI (1163) SalI (1169) AccI (1170) HincII (1171) SbfI (1179) BlpI (1244) BsaAI (1485) BstAPI (1509) XcmI (1627) EagI (1717) FspI (1913) PflFI - Tth111I (1929) BsrDI (2044) pFN2K (GST) 4132 bp
AseI  (20)
1 site
A T T A A T T A A T T A
AclI  (3972)
1 site
A A C G T T T T G C A A
BsaI  (3836)
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.
MluI  (3565)
1 site
A C G C G T T G C G C A
BspHI  (3308)
1 site
T C A T G A A G T A C T
ApaLI  (2902)
1 site
G T G C A C C A C G T G
AgeI  (2520)
1 site
A C C G G T T G G C C A
RsrII  (2327)
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  (2313)
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.
NgoMIV  (2311)
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.
EcoNI  (80)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BsgI  (356)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
XmnI  (463)
1 site
G A A N N N N T T C C T T N N N N A A G
AsiSI  (764)
1 site
G C G A T C G C C G C T A G C G
PvuI  (764)
1 site
C G A T C G G C T A G C
SgfI  (764)
1 site
G C G A T C G C C G C T A G C G
BtgI  (767)
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  (767)
1 site
C C A T G G G G T A C C
ZraI  (925)
1 site
G A C G T C C T G C A G
AatII  (927)
1 site
G A C G T C C T G C A G
BpmI  (965)
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.
DraI  (1131)
1 site
T T T A A A A A A T T T
PmeI  (1131)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (1136)
1 site
G A A T T C C T T A A G
Eco53kI  (1144)
1 site
G A G C T C C T C G A G
SacI  (1146)
1 site
G A G C T C C T C G A G
Acc65I  (1148)
1 site
G G T A C C C C A T G G
AvaI  (1152)
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  (1152)
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  (1152)
1 site
G G T A C C C C A T G G
TspMI  (1152)
1 site
C C C G G G G G G C C C
XmaI  (1152)
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  (1153)
1 site
C Y C G R G G R G C Y C
SmaI  (1154)
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  (1157)
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  (1163)
1 site
T C T A G A A G A T C T
SalI  (1169)
1 site
G T C G A C C A G C T G
AccI  (1170)
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  (1171)
1 site
G T Y R A C C A R Y T G
SbfI  (1179)
1 site
C C T G C A G G G G A C G T C C
BlpI  (1244)
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.
BsaAI  (1485)
1 site
Y A C G T R R T G C A Y
BstAPI  (1509)
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  (1627)
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.
EagI  (1717)
1 site
C G G C C G G C C G G C
FspI  (1913)
1 site
T G C G C A A C G C G T
PflFI  (1929)
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  (1929)
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  (2044)
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.
NeoR/KanR
1683 .. 2477  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
1683 .. 2477  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
GST
70 .. 723  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
70 .. 723  =  654 bp
218 amino acids  =  25.5 kDa
Product: glutathione S-transferase from Schistosoma japonicum
ori
2649 .. 3237  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2649 .. 3237  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
barnase
791 .. 1126  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus amyloliquefaciens
The barnase gene is lethal in standard bacterial transformation strains.
barnase
791 .. 1126  =  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
3353 .. 3636  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
cer region
3353 .. 3636  =  284 bp
ColE1-derived recombination site that helps to maintain plasmids as monomers
rrnB T1 terminator
3866 .. 3952  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
3866 .. 3952  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T7 terminator
1255 .. 1302  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1255 .. 1302  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
rrnB T2 terminator
4044 .. 4071  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
4044 .. 4071  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
TEV site
742 .. 762  =  21 bp
7 amino acids  =  884.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
742 .. 762  =  21 bp
7 amino acids  =  884.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
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:  70 .. 777  =  708 bp
ORF:  235 amino acids  =  27.2 kDa
ORF:  1855 .. 2241  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  3598 .. 3945  =  348 bp
ORF:  115 amino acids  =  12.4 kDa
ORF:  791 .. 1126  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  669 .. 1079  =  411 bp
ORF:  136 amino acids  =  15.5 kDa
ORF:  1683 .. 2477  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  623 .. 847  =  225 bp
ORF:  74 amino acids  =  8.6 kDa
ORF:  3707 .. 4033  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  507 .. 770  =  264 bp
ORF:  87 amino acids  =  9.7 kDa
ORF:  1992 .. 2246  =  255 bp
ORF:  84 amino acids  =  9.8 kDa
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