pCMV6-AN-mKate

PrecisionShuttle™ mammalian expression vector for fusing an ORF to an N-terminal mKate2 tag.

Sequence Author: OriGene

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HomePlasmidsMammalian Expression VectorspCMV6-AN-mKate
AanI (6553) XmnI (5875) ScaI (5756) PciI (4383) BstZ17I (4004) BsmI (3952) AanI (3891) BstBI (3714) PflFI - Tth111I (3150) NdeI (592) SnaBI (698) CMV promoter VP1.5 (forward primer) (839 .. 856) Eco53kI (924) SacI (926) T7 promoter EcoRI (979) BamHI (992) Acc65I (998) KpnI (1002) PasI (1241) PshAI (1650) BspEI (1728) AsiSI - SgfI (1744) MreI - SgrAI (1746) AscI (1750) HindIII (1764) RsrII (1781) MluI (1787) NotI (1797) PaeR7I - PspXI - XhoI (1805) PmeI (1814) FseI (1824) SacII (1828) XL39 (reverse primer) (1920 .. 1939) EcoO109I (2044) BsgI (2098) BbvCI - Bpu10I (2140) BlpI (2184) BsmBI (2204) BstXI (2291) AgeI (2402) PflMI (2405) XcmI (2497) SexAI * (2589) SfiI (2775) AvrII (2822) BclI * (2873) BsaBI * (2891) KasI (3031) NarI (3032) SfoI (3033) PluTI (3035) PstI (3085) pCMV6-AN-mKate 6625 bp
AanI  (6553)
2 sites		 T T A T A A A A T A T T
XmnI  (5875)
1 site		 G A A N N N N T T C C T T N N N N A A G
ScaI  (5756)
1 site		 A G T A C T T C A T G A
PciI  (4383)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BstZ17I  (4004)
1 site		 G T A T A C C A T A T G
BsmI  (3952)
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  (3891)
2 sites		 T T A T A A A A T A T T
BstBI  (3714)
1 site		 T T C G A A A A G C T T
PflFI  (3150)
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  (3150)
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.
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
PasI  (1241)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
PshAI  (1650)
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.
BspEI  (1728)
1 site		 T C C G G A A G G C C T
AsiSI  (1744)
1 site		 G C G A T C G C C G C T A G C G
SgfI  (1744)
1 site		 G C G A T C G C C G C T A G C G
MreI  (1746)
1 site		 C G C C G G C G G C G G C C G C
SgrAI  (1746)
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  (1750)
1 site		 G G C G C G C C C C G C G C G G
HindIII  (1764)
1 site		 A A G C T T T T C G A A
RsrII  (1781)
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  (1787)
1 site		 A C G C G T T G C G C A
NotI  (1797)
1 site		 G C G G C C G C C G C C G G C G
PaeR7I  (1805)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1805)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (1805)
1 site		 C T C G A G G A G C T C
PmeI  (1814)
1 site		 G T T T A A A C C A A A T T T G
FseI  (1824)
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  (1828)
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.
EcoO109I  (2044)
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.
BsgI  (2098)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BbvCI  (2140)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (2140)
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  (2184)
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  (2204)
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  (2291)
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  (2402)
1 site		 A C C G G T T G G C C A
PflMI  (2405)
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  (2497)
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  (2589)
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  (2775)
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.
AvrII  (2822)
1 site		 C C T A G G G G A T C C
BclI  (2873)
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  (2891)
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.
KasI  (3031)
1 site		 G G C G C C C C G C G G
NarI  (3032)
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.
SfoI  (3033)
1 site		 G G C G C C C C G C G G
PluTI  (3035)
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.
PstI  (3085)
1 site		 C T G C A G G A C G T C
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
1920 .. 1939  =  20 annealed bases
Tm  =  59°C
AmpR
5203 .. 6063  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5203 .. 5994  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5203 .. 6063  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5995 .. 6063  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5203 .. 6063  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2904 .. 3698  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2904 .. 3698  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
mKate2
1020 .. 1715  =  696 bp
232 amino acids  =  26.1 kDa
Product: monomeric far-red fluorescent protein (Shcherbo et al., 2009)
mammalian codon-optimized
mKate2
1020 .. 1715  =  696 bp
232 amino acids  =  26.1 kDa
Product: monomeric far-red fluorescent protein (Shcherbo et al., 2009)
mammalian codon-optimized
hGH poly(A) signal
1857 .. 2479  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
1857 .. 2479  =  623 bp
human growth hormone polyadenylation signal
ori
4444 .. 5032  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4444 .. 5032  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6195 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6195 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
2508 .. 2837  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
2508 .. 2837  =  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
SV40 poly(A) signal
3872 .. 3993  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3872 .. 3993  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6064 .. 6168  =  105 bp
AmpR promoter
6064 .. 6168  =  105 bp
MCS
1728 .. 1830  =  103 bp
multiple cloning site
MCS
1728 .. 1830  =  103 bp
multiple cloning site
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  3076 .. 3462  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  5333 .. 5599  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  882 .. 1775  =  894 bp
ORF:  297 amino acids  =  33.2 kDa