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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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 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 N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
PstI  (2121)
1 site
C T G C A G G A C G T C
SalI  (2123)
1 site
G T C G A C C A G C T G
PaeR7I  (2276)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2276)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2276)
1 site
C T C G A G G A G C T C
EcoNI  (2286)
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.
BspQI  (2355)
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  (2355)
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.
BsgI  (2562)
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).
BstBI  (2673)
1 site
T T C G A A A A G C T T
SwaI  (2703)
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.
BclI  (2710)
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.
PmeI  (2995)
1 site
G T T T A A A C C A A A T T T G
AmpR
4026 .. 4886  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4026 .. 4817  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4026 .. 4886  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4818 .. 4886  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4026 .. 4886  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CmR
782 .. 1441  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
782 .. 1441  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
GST
2282 .. 2929  =  648 bp
216 amino acids  =  25.3 kDa
Product: glutathione S-transferase from
Schistosoma japonicum
GST
2282 .. 2929  =  648 bp
216 amino acids  =  25.3 kDa
Product: glutathione S-transferase from
Schistosoma japonicum
ori
3267 .. 3855  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3267 .. 3855  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
17 .. 515  =  499 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
17 .. 515  =  499 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
f1 ori
5325 .. 5780  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
5325 .. 5780  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
ccdB
1783 .. 2088  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
ccdB
1783 .. 2088  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
attR1
549 .. 673  =  125 bp
recombination site for the Gateway® LR reaction
attR1
549 .. 673  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2129 .. 2253  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2129 .. 2253  =  125 bp
recombination site for the Gateway® LR reaction
AmpR promoter
4887 .. 4991  =  105 bp
AmpR promoter
4887 .. 4991  =  105 bp
T7 terminator
3020 .. 3067  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
3020 .. 3067  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac UV5 promoter
698 .. 728  =  31 bp
   Segment 1:  -35  
   698 .. 703  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
698 .. 728  =  31 bp
   Segment 2:  
   704 .. 721  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
698 .. 728  =  31 bp
   Segment 3:  -10  
   722 .. 728  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
698 .. 728  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
5948 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
5948 .. 2  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ATG
516 .. 518  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
516 .. 518  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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