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Plasmid Files

pBK-CMV

Dual eukaryotic and prokaryotic expression vector.

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pBK-CMV Sequence and MappBK-CMV.dna
Map and Sequence File   
Sequence Author:  Agilent Technologies
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 SfiI (4099) BseRI (4055) StuI (4050) PluTI (3872) SfoI (3870) NarI (3869) KasI (3868) MscI (3789) PflFI - Tth111I (3752) BsrDI (3640) RsrII (3352) BstBI (3187) PfoI (3091) BsaI (2878) ApaLI (2263) DraIII (240) MluI (463) Bts α I (516) HpaI (590) MfeI (599) BclI * (692) PvuI (852) Acc65I (1015) KpnI (1019) TspMI - XmaI (1024) AleI - SmaI (1026) BstXI (1028) PspOMI (1042) ApaI (1046) NotI - SacII (1049) XbaI (1056) ScaI (1064) PaeR7I - XhoI (1068) HindIII (1074) EcoRI (1083) BamHI (1092) SpeI (1098) SalI (1104) AccI (1105) PstI - SbfI (1114) BssHII (1116) Eco53kI (1124) SacI (1126) NheI (1300) BmtI (1304) SnaBI (1555) NdeI (1659) CMV enhancer AseI (1886) PciI (1949) pBK-CMV 4518 bp
SfiI  (4099)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
BseRI  (4055)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
StuI  (4050)
1 site
A G G C C T T C C G G A
PluTI  (3872)
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.
SfoI  (3870)
1 site
G G C G C C C C G C G G
NarI  (3869)
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.
KasI  (3868)
1 site
G G C G C C C C G C G G
MscI  (3789)
1 site
T G G C C A A C C G G T
PflFI  (3752)
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  (3752)
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.
BsrDI  (3640)
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.
RsrII  (3352)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BstBI  (3187)
1 site
T T C G A A A A G C T T
PfoI  (3091)
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.
BsaI  (2878)
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.
ApaLI  (2263)
1 site
G T G C A C C A C G T G
DraIII  (240)
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.
MluI  (463)
1 site
A C G C G T T G C G C A
BtsαI  (516)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
HpaI  (590)
1 site
G T T A A C C A A T T G
MfeI  (599)
1 site
C A A T T G G T T A A C
BclI  (692)
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.
PvuI  (852)
1 site
C G A T C G G C T A G C
Acc65I  (1015)
1 site
G G T A C C C C A T G G
KpnI  (1019)
1 site
G G T A C C C C A T G G
TspMI  (1024)
1 site
C C C G G G G G G C C C
XmaI  (1024)
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.
AleI  (1026)
1 site
C A C N N N N G T G G T G N N N N C A C
SmaI  (1026)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BstXI  (1028)
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.
PspOMI  (1042)
1 site
G G G C C C C C C G G G
ApaI  (1046)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
NotI  (1049)
1 site
G C G G C C G C C G C C G G C G
SacII  (1049)
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.
XbaI  (1056)
1 site
T C T A G A A G A T C T
ScaI  (1064)
1 site
A G T A C T T C A T G A
PaeR7I  (1068)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1068)
1 site
C T C G A G G A G C T C
HindIII  (1074)
1 site
A A G C T T T T C G A A
EcoRI  (1083)
1 site
G A A T T C C T T A A G
BamHI  (1092)
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.
SpeI  (1098)
1 site
A C T A G T T G A T C A
SalI  (1104)
1 site
G T C G A C C A G C T G
AccI  (1105)
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.
PstI  (1114)
1 site
C T G C A G G A C G T C
SbfI  (1114)
1 site
C C T G C A G G G G A C G T C C
BssHII  (1116)
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.
Eco53kI  (1124)
1 site
G A G C T C C T C G A G
SacI  (1126)
1 site
G A G C T C C T C G A G
NheI  (1300)
1 site
G C T A G C C G A T C G
BmtI  (1304)
1 site
G C T A G C C G A T C G
SnaBI  (1555)
1 site
T A C G T A A T G C A T
NdeI  (1659)
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.
AseI  (1886)
1 site
A T T A A T T A A T T A
PciI  (1949)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NeoR/KanR
3206 .. 4000  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
3206 .. 4000  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
2010 .. 2598  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2010 .. 2598  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
7 .. 462  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
7 .. 462  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
lacZα
797 .. 1183  =  387 bp
128 amino acids  =  14.7 kDa
Product: LacZα fragment of β-galactosidase
lacZα
797 .. 1183  =  387 bp
128 amino acids  =  14.7 kDa
Product: LacZα fragment of β-galactosidase
SV40 promoter
4035 .. 4392  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
4035 .. 4392  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
1532 .. 1835  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1532 .. 1835  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
1328 .. 1531  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
1328 .. 1531  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
469 .. 590  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
469 .. 590  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4394 .. 4498  =  105 bp
AmpR promoter
4394 .. 4498  =  105 bp
HSV TK poly(A) signal
2927 .. 2974  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
2927 .. 2974  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
lac operator
1203 .. 1219  =  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
1203 .. 1219  =  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).
SV40 ori
4049 .. 4184  =  136 bp
SV40 origin of replication
SV40 ori
4049 .. 4184  =  136 bp
SV40 origin of replication
MCS
1015 .. 1127  =  113 bp
multiple cloning site
MCS
1015 .. 1127  =  113 bp
multiple cloning site
T7 promoter
975 .. 993  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
975 .. 993  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
1140 .. 1158  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
1140 .. 1158  =  19 bp
promoter for bacteriophage T3 RNA polymerase
M13 fwd
952 .. 968  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
952 .. 968  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
1179 .. 1195  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
1179 .. 1195  =  17 bp
common sequencing primer, one of multiple similar
variants
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