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Plasmid Files

pGFP

Vector for expressing GFP in bacteria.

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pGFP Sequence and MappGFP.dna
Map and Sequence File   
Sequence Author:  Clontech
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 BspQI - SapI (3336) PciI (3219) AhdI (2331) BmrI (2291) NmeAIII (2184) FspI (2108) PvuI (1962) TsoI (1933) ScaI (1850) XmnI (1731) HindIII (234) BfuAI - BspMI (239) SphI (244) PstI - SbfI (250) SalI (252) XbaI (258) BamHI (264) AvaI - BsoBI - TspMI - XmaI (269) BmeT110I (270) SmaI (271) Acc65I (273) AgeI (276) KpnI (277) BtgI - NcoI - StyI (454) MscI (459) BsrGI (564) BsaAI - PmlI (613) BstZ17I (738) HpaI (777) MfeI (848) BtgZI (869) BstBI (910) BsmI (1005) EcoRI (1014) StuI (1074) SpeI (1078) EagI (1084) BsiWI (1088) PspOMI (1093) ApaI - BanII (1097) PfoI (1151) BstAPI (1290) AflII (1346) ZraI (1410) AatII (1412) SspI (1526) pGFP 3344 bp
BspQI  (3336)
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  (3336)
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.
PciI  (3219)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (2331)
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.
BmrI  (2291)
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.
NmeAIII  (2184)
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).
FspI  (2108)
1 site
T G C G C A A C G C G T
PvuI  (1962)
1 site
C G A T C G G C T A G C
TsoI  (1933)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
ScaI  (1850)
1 site
A G T A C T T C A T G A
XmnI  (1731)
1 site
G A A N N N N T T C C T T N N N N A A G
HindIII  (234)
1 site
A A G C T T T T C G A A
BfuAI  (239)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (239)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
SphI  (244)
1 site
G C A T G C C G T A C G
PstI  (250)
1 site
C T G C A G G A C G T C
SbfI  (250)
1 site
C C T G C A G G G G A C G T C C
SalI  (252)
1 site
G T C G A C C A G C T G
XbaI  (258)
1 site
T C T A G A A G A T C T
BamHI  (264)
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  (269)
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  (269)
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  (269)
1 site
C C C G G G G G G C C C
XmaI  (269)
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  (270)
1 site
C Y C G R G G R G C Y C
SmaI  (271)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (273)
1 site
G G T A C C C C A T G G
AgeI  (276)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
KpnI  (277)
1 site
G G T A C C C C A T G G
BtgI  (454)
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  (454)
1 site
C C A T G G G G T A C C
StyI  (454)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
MscI  (459)
1 site
T G G C C A A C C G G T
BsrGI  (564)
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.
BsaAI  (613)
1 site
Y A C G T R R T G C A Y
PmlI  (613)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BstZ17I  (738)
1 site
G T A T A C C A T A T G
HpaI  (777)
1 site
G T T A A C C A A T T G
MfeI  (848)
1 site
C A A T T G G T T A A C
BtgZI  (869)
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  (910)
1 site
T T C G A A A A G C T T
BsmI  (1005)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
EcoRI  (1014)
1 site
G A A T T C C T T A A G
StuI  (1074)
1 site
A G G C C T T C C G G A
SpeI  (1078)
1 site
A C T A G T T G A T C A
EagI  (1084)
1 site
C G G C C G G C C G G C
BsiWI  (1088)
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.
PspOMI  (1093)
1 site
G G G C C C C C C G G G
ApaI  (1097)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (1097)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PfoI  (1151)
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.
BstAPI  (1290)
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.
AflII  (1346)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
ZraI  (1410)
1 site
G A C G T C C T G C A G
AatII  (1412)
1 site
G A C G T C C T G C A G
SspI  (1526)
1 site
A A T A T T T T A T A A
AmpR
1544 .. 2404  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1544 .. 1612  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1544 .. 2404  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1613 .. 2404  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1544 .. 2404  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
GFP
289 .. 1005  =  717 bp
238 amino acids  =  26.9 kDa
Product: Aequoria victoria green fluorescent protein
GFP
289 .. 1005  =  717 bp
238 amino acids  =  26.9 kDa
Product: Aequoria victoria green fluorescent protein
ori
2575 .. 3163  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2575 .. 3163  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
1439 .. 1543  =  105 bp
AmpR promoter
1439 .. 1543  =  105 bp
lac promoter
143 .. 173  =  31 bp
   Segment 1:  -35  
   143 .. 148  =  6 bp
promoter for the E. coli lac operon
lac promoter
143 .. 173  =  31 bp
   Segment 2:  
   149 .. 166  =  18 bp
promoter for the E. coli lac operon
lac promoter
143 .. 173  =  31 bp
   Segment 3:  -10  
   167 .. 173  =  7 bp
promoter for the E. coli lac operon
lac promoter
143 .. 173  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
181 .. 197  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
181 .. 197  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
M13 rev
205 .. 221  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
205 .. 221  =  17 bp
common sequencing primer, one of multiple similar
variants
ATG
217 .. 219  =  3 bp
1 amino acid  =  149.2 Da
Product: lacZ start codon
ATG
217 .. 219  =  3 bp
1 amino acid  =  149.2 Da
Product: lacZ start codon
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