Resources
Plasmid Files

pFA6a-GFP(S65T)-kanMX6

Plasmid with a kanMX marker for adding a C-terminal GFP tag or truncating a gene.

 
To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pFA6a-GFP(S65T)-kanMX6.dna
Map and Sequence File:    Download    Open   
Download Free Trial Get SnapGene Viewer


BsiWI (25) PfoI (4633) EcoO109I (4576) AatII (4522) ZraI (4520) XmnI (4199) NmeAIII (3748) BmrI (3640) BanI (3548) PciI (2707) BspQI - SapI (2591) SacII (2476) SfiI (2469) SpeI (2456) EcoRV (2446) R1 (2418 .. 2437) EcoRI (2432) SacI (2430) Eco53kI (2428) SalI (37) F2 (42 .. 61) BamHI (43) AvaI - BsoBI - TspMI - XmaI (48) BmeT110I (49) SmaI (50) PacI (58) MscI (230) SnaBI (253) BsrGI (335) PmlI (384) MfeI (619) BtgZI (640) BstBI (681) F3 (777 .. 793) AscI - BssHII (778) PsiI (839) BglII (986) BstEII (1016) BstXI (1033) BmgBI (1069) BseRI (1180) MluI (1233) NruI (1457) EcoNI (1712) AsiSI (1800) PflMI (2063) PmeI (2421) pFA6a-GFP(S65T)-kanMX6 4854 bp
BsiWI  (25)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
PfoI  (4633)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
EcoO109I  (4576)
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.
AatII  (4522)
1 site
G A C G T C C T G C A G
ZraI  (4520)
1 site
G A C G T C C T G C A G
XmnI  (4199)
1 site
G A A N N N N T T C C T T N N N N A A G
NmeAIII  (3748)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BmrI  (3640)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
BanI  (3548)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
PciI  (2707)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2591)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (2591)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
SacII  (2476)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
SfiI  (2469)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
SpeI  (2456)
1 site
A C T A G T T G A T C A
EcoRV  (2446)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
EcoRI  (2432)
1 site
G A A T T C C T T A A G
SacI  (2430)
1 site
G A G C T C C T C G A G
Eco53kI  (2428)
1 site
G A G C T C C T C G A G
SalI  (37)
1 site
G T C G A C C A G C T G
BamHI  (43)
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.
AvaI  (48)
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  (48)
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  (48)
1 site
C C C G G G G G G C C C
XmaI  (48)
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  (49)
1 site
C Y C G R G G R G C Y C
SmaI  (50)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PacI  (58)
1 site
T T A A T T A A A A T T A A T T
MscI  (230)
1 site
T G G C C A A C C G G T
SnaBI  (253)
1 site
T A C G T A A T G C A T
BsrGI  (335)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
PmlI  (384)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
MfeI  (619)
1 site
C A A T T G G T T A A C
BtgZI  (640)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BstBI  (681)
1 site
T T C G A A A A G C T T
AscI  (778)
1 site
G G C G C G C C C C G C G C G G
BssHII  (778)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
PsiI  (839)
1 site
T T A T A A A A T A T T
BglII  (986)
1 site
A G A T C T T C T A G A
BstEII  (1016)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BstXI  (1033)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BmgBI  (1069)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
BseRI  (1180)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
MluI  (1233)
1 site
A C G C G T T G C G C A
NruI  (1457)
1 site
T C G C G A A G C G C T
EcoNI  (1712)
1 site
C C T N N N N N A G G G G A