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Plasmid Files

pANT7_cGST

Gateway® destination vector for in vitro transcription and translation of a protein with a C-terminal GST tag.

 
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pANT7_cGST.dna
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PspOMI (55) NaeI (5652) NgoMIV (5650) PsiI (5421) PfoI (5132) AatII (5021) ZraI (5019) NmeAIII (4247) AhdI (4099) PspFI (3514) BseYI (3510) BlpI (3009) SpeI (3000) ApaI (59) AvrII (93) PmlI (258) AarI (281) Acc65I (383) KpnI (387) Eco53kI (537) SacI (539) BspEI (991) PasI (1227) NcoI (1296) BssHII (1489) BstZ17I (1530) BbvCI (1720) TspMI - XmaI (1864) SmaI - SrfI (1866) BstXI (1983) PstI (2121) SalI (2123) PaeR7I - PspXI - XhoI (2276) EcoNI (2286) BspQI - SapI (2355) BsgI (2562) BstBI (2673) SwaI (2703) BclI * (2710) PmeI (2995) pANT7_cGST 5964 bp
PspOMI  (55)
1 site
G G G C C C C C C G G G
NaeI  (5652)
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  (5650)
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.
PsiI  (5421)
1 site
T T A T A A A A T A T T
PfoI  (5132)
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.
AatII  (5021)
1 site
G A C G T C C T G C A G
ZraI  (5019)
1 site
G A C G T C C T G C A G
NmeAIII  (4247)
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).
AhdI  (4099)
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.
PspFI  (3514)
1 site
C C C A G C G G G T C G
BseYI  (3510)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
BlpI  (3009)
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.
SpeI  (3000)
1 site
A C T A G T T G A T C A
ApaI  (59)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AvrII  (93)
1 site
C C T A G G G G A T C C
PmlI  (258)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AarI  (281)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
Acc65I  (383)
1 site
G G T A C C C C A T G G
KpnI  (387)
1 site
G G T A C C C C A T G G
Eco53kI  (537)
1 site
G A G C T C C T C G A G
SacI  (539)
1 site
G A G C T C C T C G A G
BspEI  (991)
1 site
T C C G G A A G G C C T
PasI  (1227)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
NcoI  (1296)
1 site
C C A T G G G G T A C C
BssHII  (1489)
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.
BstZ17I  (1530)
1 site
G T A T A C C A T A T G
BbvCI  (1720)
1 site
C C T C A G C G G A G T C G
TspMI  (1864)
1 site
C C C G G G G G G C C C
XmaI  (1864)
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.
SmaI  (1866)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (1866)
1 site
G C C C G G G C C G G G C C C G
BstXI  (1983)
1 site
C C A N N N N N N T G G G G T