pPCR-Script Cam SK(+)

Bacterial cloning vector with a chloramphenicol resistance marker and an SrfI site for inserting blunt PCR products.

Sequence Author: Agilent Technologies

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XmnI (3083) BclI * (3057) Bsu36I (2993) AgeI (2943) PflFI - Tth111I (2886) BstBI (2869) ScaI (2755) TatI (2753) NcoI - StyI (2639) MscI (2605) BsrDI (2357) SnaBI (2344) BspEI (2334) PsiI (99) DraIII (227) NgoMIV (328) NaeI (330) BglI (472) FspI (479) PvuI (500) BmrI (580) Acc65I (653) KpnI (657) PspOMI (659) EcoO109I (660) ApaI (663) AbsI - PaeR7I - PspXI - XhoI (668) SalI (674) AccI (675) HincII (676) BspDI - ClaI (684) HindIII (689) EcoRV (697) EcoRI (701) PstI (711) BamHI (719) SrfI (728) EagI - NotI (738) AleI (749) SacII (750) BstXI (751) Eco53kI (757) SacI (759) lac operator BspQI - SapI (1037) AflIII - PciI (1153) NspI (1157) BseYI (1457) PspFI (1461) AlwNI (1569) pPCR-Script Cam SK(+) 3400 bp
XmnI  (3083)
1 site
G A A N N N N T T C C T T N N N N A A G
BclI  (3057)
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.
Bsu36I  (2993)
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.
AgeI  (2943)
1 site
A C C G G T T G G C C A
PflFI  (2886)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2886)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BstBI  (2869)
1 site
T T C G A A A A G C T T
ScaI  (2755)
1 site
A G T A C T T C A T G A
TatI  (2753)
1 site
W G T A C W W C A T G W
NcoI  (2639)
1 site
C C A T G G G G T A C C
StyI  (2639)
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  (2605)
1 site
T G G C C A A C C G G T
BsrDI  (2357)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
SnaBI  (2344)
1 site
T A C G T A A T G C A T
BspEI  (2334)
1 site
T C C G G A A G G C C T
PsiI  (99)
1 site
T T A T A A A A T A T T
DraIII  (227)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
NgoMIV  (328)
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.
NaeI  (330)
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.
BglI  (472)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
FspI  (479)
1 site
T G C G C A A C G C G T
PvuI  (500)
1 site
C G A T C G G C T A G C
BmrI  (580)
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.
Acc65I  (653)
1 site
G G T A C C C C A T G G
KpnI  (657)
1 site
G G T A C C C C A T G G
PspOMI  (659)
1 site
G G G C C C C C C G G G
EcoO109I  (660)
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.
ApaI  (663)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AbsI  (668)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (668)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (668)
1 site
V C T C G A G B B G A G C T C V
XhoI  (668)
1 site
C T C G A G G A G C T C
SalI  (674)
1 site
G T C G A C C A G C T G
AccI  (675)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
HincII  (676)
1 site
G T Y R A C C A R Y T G
BspDI  (684)
1 site
A T C G A T T A G C T A
ClaI  (684)
1 site
A T C G A T T A G C T A
HindIII  (689)
1 site
A A G C T T T T C G A A
EcoRV  (697)
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  (701)
1 site
G A A T T C C T T A A G
PstI  (711)
1 site
C T G C A G G A C G T C
BamHI  (719)
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.
SrfI  (728)
1 site
G C C C G G G C C G G G C C C G
EagI  (738)
1 site
C G G C C G G C C G G C
NotI  (738)
1 site
G C G G C C G C C G C C G G C G
AleI  (749)
1 site
C A C N N N N G T G G T G N N N N C A C
SacII  (750)
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.
BstXI  (751)
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.
Eco53kI  (757)
1 site
G A G C T C C T C G A G
SacI  (759)
1 site
G A G C T C C T C G A G
BspQI  (1037)
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  (1037)
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  (1153)
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  (1153)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (1157)
1 site
R C A T G Y Y G T A C R
BseYI  (1457)
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.
PspFI  (1461)
1 site
C C C A G C G G G T C G
AlwNI  (1569)
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.
CmR
2125 .. 2784  =  660 bp
219 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2125 .. 2784  =  660 bp
219 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
1214 .. 1802  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1214 .. 1802  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
lacZα
241 .. 816  =  576 bp
191 amino acids  =  20.4 kDa
Product: LacZα fragment of β-galactosidase
lacZα
241 .. 816  =  576 bp
191 amino acids  =  20.4 kDa
Product: LacZα fragment of β-galactosidase
AmpR promoter
3272 .. 3377  =  106 bp
AmpR promoter
3272 .. 3377  =  106 bp
cat promoter
2022 .. 2124  =  103 bp
promoter of the E. coli cat gene
cat promoter
2022 .. 2124  =  103 bp
promoter of the E. coli cat gene
lac promoter
860 .. 890  =  31 bp
3 segments
   Segment 3:  -10  
   860 .. 866  =  7 bp
promoter for the E. coli lac operon
lac promoter
860 .. 890  =  31 bp
3 segments
   Segment 2:  
   867 .. 884  =  18 bp
promoter for the E. coli lac operon
lac promoter
860 .. 890  =  31 bp
3 segments
   Segment 1:  -35  
   885 .. 890  =  6 bp
promoter for the E. coli lac operon
lac promoter
860 .. 890  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
836 .. 852  =  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
836 .. 852  =  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).
f1 ori
3 .. 458  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3 .. 458  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
MCS
653 .. 760  =  108 bp
multiple cloning site
MCS
653 .. 760  =  108 bp
multiple cloning site
T7 promoter
626 .. 644  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
626 .. 644  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
773 .. 791  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
773 .. 791  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
600 .. 616  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
600 .. 616  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
812 .. 828  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
812 .. 828  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  2125 .. 2784  =  660 bp
ORF:  219 amino acids  =  25.6 kDa
ORF:  3008 .. 3271  =  264 bp
ORF:  87 amino acids  =  9.9 kDa
ORF:  241 .. 816  =  576 bp
ORF:  191 amino acids  =  20.4 kDa
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