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Plasmid Files

pCMV6-A-BSD

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

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pCMV6-A-BSD Sequence and MappCMV6-A-BSD.dna
Map and Sequence File   
Sequence Author:  OriGene
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 DraIII (5308) XmnI (4755) ScaI (4636) AlwNI (3679) PciI (3263) BspQI - SapI (3147) BstZ17I (2884) BsmI (2832) AanI (2771) AanI (5433) BsrGI (301) 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
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
AanI  (5433)
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

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
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

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
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
   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
   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
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