pCMV6-A-BSD

PrecisionShuttle™ mammalian vector with a blasticidin resistance marker, for expressing an untagged ORF.

Sequence Author: OriGene

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HomePlasmidsMammalian Expression VectorspCMV6-A-BSD
AanI (5433) DraIII (5308) XmnI (4755) ScaI (4636) AlwNI (3679) PciI (3263) BspQI - SapI (3147) BstZ17I (2884) BsmI (2832) AanI (2771) BsrGI (301) CMV enhancer NdeI (592) SnaBI (698) VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) AsiSI - SgfI (1024) MreI - SgrAI (1026) AscI - BssHII (1030) HindIII (1044) RsrII (1061) MluI (1067) NotI (1077) PaeR7I - PspXI - XhoI (1085) PmeI (1094) FseI (1104) SacII (1108) AleI (1194) XL39 (reverse primer) (1200 .. 1219) BbvCI - Bpu10I (1420) BlpI (1464) BsmBI (1484) BstXI (1571) AgeI (1682) PflMI (1685) XcmI (1777) SexAI * (1869) SfiI (2055) StuI (2101) AvrII (2102) BclI * (2153) BsiWI (2166) PpuMI (2334) NruI * (2406) BsaBI * (2473) PstI (2578) BstBI (2594) BfuAI - BspMI (2630) PfoI (2687) pCMV6-A-BSD 5505 bp
AanI  (5433)
2 sites		 T T A T A A A A T A T T
DraIII  (5308)
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.
XmnI  (4755)
1 site		 G A A N N N N T T C C T T N N N N A A G
ScaI  (4636)
1 site		 A G T A C T T C A T G A
AlwNI  (3679)
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.
PciI  (3263)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (3147)
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  (3147)
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.
BstZ17I  (2884)
1 site		 G T A T A C C A T A T G
BsmI  (2832)
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.
AanI  (2771)
2 sites		 T T A T A A A A T A T T
BsrGI  (301)
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.
NdeI  (592)
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.
SnaBI  (698)
1 site		 T A C G T A A T G C A T
Eco53kI  (924)
1 site		 G A G C T C C T C G A G
SacI  (926)
1 site		 G A G C T C C T C G A G
EcoRI  (979)
1 site		 G A A T T C C T T A A G
BamHI  (992)
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.
Acc65I  (998)
1 site		 G G T A C C C C A T G G
KpnI  (1002)
1 site		 G G T A C C C C A T G G
AsiSI  (1024)
1 site		 G C G A T C G C C G C T A G C G
SgfI  (1024)
1 site		 G C G A T C G C C G C T A G C G
MreI  (1026)
1 site		 C G C C G G C G G C G G C C G C
SgrAI  (1026)
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.
AscI  (1030)
1 site		 G G C G C G C C C C G C G C G G
BssHII  (1030)
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.
HindIII  (1044)
1 site		 A A G C T T T T C G A A
RsrII  (1061)
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.
MluI  (1067)
1 site		 A C G C G T T G C G C A
NotI  (1077)
1 site		 G C G G C C G C C G C C G G C G
PaeR7I  (1085)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1085)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (1085)
1 site		 C T C G A G G A G C T C
PmeI  (1094)
1 site		 G T T T A A A C C A A A T T T G
FseI  (1104)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
SacII  (1108)
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.
AleI  (1194)
1 site		 C A C N N N N G T G G T G N N N N C A C
BbvCI  (1420)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (1420)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BlpI  (1464)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
BsmBI  (1484)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
BstXI  (1571)
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.
AgeI  (1682)
1 site		 A C C G G T T G G C C A
PflMI  (1685)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
XcmI  (1777)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
SexAI  (1869)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SfiI  (2055)
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.
StuI  (2101)
1 site		 A G G C C T T C C G G A
AvrII  (2102)
1 site		 C C T A G G G G A T C C
BclI  (2153)
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.
BsiWI  (2166)
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.
PpuMI  (2334)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
NruI  (2406)
1 site		 T C G C G A A G C G C T
* Blocked by Dam methylation.
BsaBI  (2473)
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.
PstI  (2578)
1 site		 C T G C A G G A C G T C
BstBI  (2594)
1 site		 T T C G A A A A G C T T
BfuAI  (2630)
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  (2630)
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.
PfoI  (2687)
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.
VP1.5 (forward primer)
18-mer  /  44% GC
1 binding site
839 .. 856  =  18 annealed bases
Tm  =  51°C
XL39 (reverse primer)
20-mer  /  55% GC
1 binding site
1200 .. 1219  =  20 annealed bases
Tm  =  59°C
AmpR
4083 .. 4943  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4083 .. 4874  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4083 .. 4943  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4875 .. 4943  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4083 .. 4943  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
hGH poly(A) signal
1137 .. 1759  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
1137 .. 1759  =  623 bp
human growth hormone polyadenylation signal
ori
3324 .. 3912  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3324 .. 3912  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5075 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5075 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
2175 .. 2573  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
2175 .. 2573  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
1788 .. 2117  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1788 .. 2117  =  330 bp
SV40 enhancer and early promoter
CMV promoter
723 .. 926  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
723 .. 926  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
MCS
979 .. 1110  =  132 bp
multiple cloning site
MCS
979 .. 1110  =  132 bp
multiple cloning site
SV40 poly(A) signal
2752 .. 2873  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2752 .. 2873  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4944 .. 5048  =  105 bp
AmpR promoter
4944 .. 5048  =  105 bp
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
1968 .. 2103  =  136 bp
SV40 origin of replication
SV40 ori
1968 .. 2103  =  136 bp
SV40 origin of replication
ORF:  2509 .. 2787  =  279 bp
ORF:  92 amino acids  =  10.4 kDa
ORF:  4213 .. 4479  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2175 .. 2573  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  1501 .. 1725  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  2103 .. 2477  =  375 bp
ORF:  124 amino acids  =  13.5 kDa
ORF:  4083 .. 4943  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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