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Plasmid Files

YIplac204

Yeast integrating plasmid with a TRP1 marker.

 
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YIplac204.dna
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BspQI - SapI (3536) AflIII - PciI (3419) AlwNI (3010) AhdI (2531) BsaI (2465) BpmI (2462) BsrFI (2446) NmeAIII (2384) TsoI (2133) ScaI (2050) XmnI (1931) HindIII (233) SphI (243) PstI - SbfI (249) SalI (251) HincII (253) XbaI (257) BamHI (263) TspMI - XmaI (268) SmaI (270) Acc65I (272) KpnI (276) Eco53kI (280) SacI (282) EcoRI (284) KasI (445) NarI (446) SfoI (447) PluTI (449) NdeI (498) PfoI (633) AarI (1012) BsgI (1038) BstXI (1155) EcoRV (1164) Bsu36I (1204) MfeI (1305) BstZ17I (1482) PmlI (1489) ZraI (1610) AatII (1612) SspI (1726) YIplac204 3545 bp
BspQI  (3536)
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  (3536)
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.
AflIII  (3419)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (3419)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (3010)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AhdI  (2531)
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.
BsaI  (2465)
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.
BpmI  (2462)
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.
BsrFI  (2446)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
NmeAIII  (2384)
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).
TsoI  (2133)
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  (2050)
1 site
A G T A C T T C A T G A
XmnI  (1931)
1 site
G A A N N N N T T C C T T N N N N A A G
HindIII  (233)
1 site
A A G C T T T T C G A A
SphI  (243)
1 site
G C A T G C C G T A C G
PstI  (249)
1 site
C T G C A G G A C G T C
SbfI  (249)
1 site
C C T G C A G G G G A C G T C C
SalI  (251)
1 site
G T C G A C C A G C T G
HincII  (253)
1 site
G T Y R A C C A R Y T G
XbaI  (257)
1 site
T C T A G A A G A T C T
BamHI  (263)
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.
TspMI  (268)
1 site
C C C G G G G G G C C C
XmaI  (268)
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  (270)
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  (272)
1 site
G G T A C C C C A T G G
KpnI  (276)
1 site
G G T A C C C C A T G G
Eco53kI  (280)
1 site
G A G C T C C T C G A G
SacI  (282)
1 site
G A G C T C C T C G A G
EcoRI  (284)
1 site
G A A T T C C T T A A G
KasI  (445)
1 site
G G C G C C C C G C G G
NarI  (446)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (447)
1 site
G G C G C C C C G C G G
PluTI  (449)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
NdeI  (498)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
PfoI  (633)
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.
AarI  (1012)
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.
BsgI  (1038)
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).
BstXI  (1155)
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.
EcoRV  (1164)
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.
Bsu36I  (1204)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
MfeI  (1305)
1 site
C A A T T G G T T A A C
BstZ17I  (1482)
1 site
G T A T A C C A T A T G
PmlI  (1489)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
ZraI  (1610)
1 site
G A C G T C C T G C A G
AatII  (1612)
1 site
G A C G T C C T G C A G
SspI  (1726)
1 site
A A T A T T T T A T A A
AmpR
1744 .. 2604  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1744 .. 1812  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1744 .. 2604  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1813 .. 2604  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1744 .. 2604  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
TRP1
775 .. 1449  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
TRP1
775 .. 1449  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis
yeast auxotrophic marker
ori
2775 .. 3363  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
2775 .. 3363  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
lacZα
216 .. 539  =  324 bp
107 amino acids  =  12.2 kDa
Product: LacZα fragment of β-galactosidase
lacZα
216 .. 539  =  324 bp
107 amino acids  =  12.2 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
1639 .. 1743  =  105 bp
AmpR promoter
1639 .. 1743  =  105 bp
TRP1 promoter
1450 .. 1551  =  102 bp
TRP1 promoter
1450 .. 1551  =  102 bp
lac promoter
142 .. 172  =  31 bp
   Segment 1:  -35  
   142 .. 147  =  6 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
   Segment 2:  
   148 .. 165  =  18 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
   Segment 3:  -10  
   166 .. 172  =  7 bp
promoter for the E. coli lac operon
lac promoter
142 .. 172  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
180 .. 196  =  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
180 .. 196  =  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).
ARS1
696 .. 936  =  241 bp
S. cerevisiae autonomously replicating sequence ARS1/ARS416
ARS1
696 .. 936  =  241 bp
S. cerevisiae autonomously replicating sequence ARS1/ARS416
MCS
233 .. 289  =  57 bp
pUC19 multiple cloning site
MCS
233 .. 289  =  57 bp
pUC19 multiple cloning site
M13 rev
204 .. 220  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
204 .. 220  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
290 .. 306  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
290 .. 306  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1744 .. 2604  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  704 .. 1039  =  336 bp
ORF:  111 amino acids  =  12.2 kDa
ORF:  216 .. 539  =  324 bp
ORF:  107 amino acids  =  12.2 kDa
ORF:  918 .. 1199  =  282 bp
ORF:  93 amino acids  =  10.8 kDa
ORF:  2208 .. 2474  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  143 .. 499  =  357 bp
ORF:  118 amino acids  =  13.5 kDa
ORF:  775 .. 1449  =  675 bp
ORF:  224 amino acids  =  24.1 kDa
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