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pSTV28

Bacterial cloning vector with a p15A origin and a chloramphenicol resistance gene. The MCS is reversed in pSTV29.

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pSTV28.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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BsrDI (2987) AclI (2915) MscI (2737) NcoI - StyI (2699) ScaI (2587) TatI (2585) BaeGI - Bme1580I (2537) DrdI - PflFI - Tth111I (2456) Bsu36I (2347) BclI * (2296) BsaBI * (2200) EcoO109I (2143) NaeI (2114) NgoMIV (2112) BbsI (2073) PshAI (2059) EcoRI (1738) BanII - SacI (1736) Eco53kI (1734) KpnI (1730) Acc65I (1726) SmaI (1724) BmeT110I (1723) AvaI - BsoBI - TspMI - XmaI (1722) BamHI (1717) HincII (1707) SalI (1705) PstI - SbfI (1703) SfcI (1699) SphI (1697) BfuAI - BspMI (1692) HindIII (1687) BmrI (1647) PvuI (1567) FspI (1546) BglI (1539) BspDI - ClaI (1518) BsaAI (312) NheI (583) BmtI (587) BstZ17I (596) XmnI (639) SgrAI (669) PfoI * (788) SacII (835) BssS α I (959) AcuI (1484) pSTV28 2999 bp
BsrDI  (2987)
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.
AclI  (2915)
1 site
A A C G T T T T G C A A
MscI  (2737)
1 site
T G G C C A A C C G G T
NcoI  (2699)
1 site
C C A T G G G G T A C C
StyI  (2699)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
ScaI  (2587)
1 site
A G T A C T T C A T G A
TatI  (2585)
1 site
W G T A C W W C A T G W
BaeGI  (2537)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (2537)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
DrdI  (2456)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PflFI  (2456)
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  (2456)
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.
Bsu36I  (2347)
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.
BclI  (2296)
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.
BsaBI  (2200)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
EcoO109I  (2143)
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.
NaeI  (2114)
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  (2112)
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.
BbsI  (2073)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
PshAI  (2059)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
EcoRI  (1738)
1 site
G A A T T C C T T A A G
BanII  (1736)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (1736)
1 site
G A G C T C C T C G A G
Eco53kI  (1734)
1 site
G A G C T C C T C G A G
KpnI  (1730)
1 site
G G T A C C C C A T G G
Acc65I  (1726)
1 site
G G T A C C C C A T G G
SmaI  (1724)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BmeT110I  (1723)
1 site
C Y C G R G G R G C Y C
AvaI  (1722)
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  (1722)
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  (1722)
1 site
C C C G G G G G G C C C
XmaI  (1722)
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.
BamHI  (1717)
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.
HincII  (1707)
1 site
G T Y R A C C A R Y T G
SalI  (1705)
1 site
G T C G A C C A G C T G
PstI  (1703)
1 site
C T G C A G G A C G T C
SbfI  (1703)
1 site
C C T G C A G G G G A C G T C C
SfcI  (1699)
1 site
C T R Y A G G A Y R T C

Sticky ends from different SfcI sites may not be compatible.
SfcI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
SphI  (1697)
1 site
G C A T G C C G T A C G
BfuAI  (1692)
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  (1692)
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.
HindIII  (1687)
1 site
A A G C T T T T C G A A
BmrI  (1647)
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.
PvuI  (1567)
1 site
C G A T C G G C T A G C
FspI  (1546)
1 site
T G C G C A A C G C G T
BglI  (1539)
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.
BspDI  (1518)
1 site
A T C G A T T A G C T A
ClaI  (1518)
1 site
A T C G A T T A G C T A
BsaAI  (312)
1 site
Y A C G T R R T G C A Y
NheI  (583)
1 site
G C T A G C C G A T C G
BmtI  (587)
1 site
G C T A G C C G A T C G
BstZ17I  (596)
1 site
G T A T A C C A T A T G
XmnI  (639)
1 site
G A A N N N N T T C C T T N N N N A A G
SgrAI  (669)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
PfoI  (788)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
SacII  (835)
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.
BssSαI  (959)
1 site
C A C G A G G T G C T C
AcuI  (1484)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
CmR
2559 .. 219  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2559 .. 219  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
p15A ori
848 .. 1392  =  545 bp
Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin.
p15A ori
848 .. 1392  =  545 bp
Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin.
lacZα
1527 .. 1757  =  231 bp
76 amino acids  =  8.5 kDa
Product: LacZα fragment of β-galactosidase
lacZα
1527 .. 1757  =  231 bp
76 amino acids  =  8.5 kDa
Product: LacZα fragment of β-galactosidase
cat promoter
220 .. 322  =  103 bp
promoter of the E. coli cat gene
cat promoter
220 .. 322  =  103 bp
promoter of the E. coli cat gene
lac promoter
1801 .. 1831  =  31 bp
   Segment 3:  -10  
   1801 .. 1807  =  7 bp
promoter for the E. coli lac operon
lac promoter
1801 .. 1831  =  31 bp
   Segment 2:  
   1808 .. 1825  =  18 bp
promoter for the E. coli lac operon
lac promoter
1801 .. 1831  =  31 bp
   Segment 1:  -35  
   1826 .. 1831  =  6 bp
promoter for the E. coli lac operon
lac promoter
1801 .. 1831  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
1777 .. 1793  =  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
1777 .. 1793  =  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).
MCS
1687 .. 1743  =  57 bp
pUC18/19 multiple cloning site
MCS
1687 .. 1743  =  57 bp
pUC18/19 multiple cloning site
M13 fwd
1667 .. 1683  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
1667 .. 1683  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
1753 .. 1769  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
1753 .. 1769  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  763 .. 1041  =  279 bp
ORF:  92 amino acids  =  10.4 kDa
ORF:  1846 .. 2262  =  417 bp
ORF:  138 amino acids  =  14.9 kDa
ORF:  1527 .. 1757  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  2559 .. 219  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  730 .. 1002  =  273 bp
ORF:  90 amino acids  =  9.7 kDa
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