pCMV6-AC-YFP

PrecisionShuttle™ mammalian expression vector for fusing an ORF to a C-terminal TurboYFP tag.

Sequence Author: OriGene

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AanI (6559) DraIII (6434) ScaI (5762) AlwNI (4808) PciI (4392) BstZ17I (4013) BsmI (3961) AanI (3900) PfoI (3816) BstBI (3723) PflFI - Tth111I (3159) 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 (1026) AscI (1030) HindIII (1044) RsrII (1061) MluI (1067) BsiWI (1070) NotI (1076) PaeR7I - XhoI (1082) BspEI (1802) PmeI (1823) FseI (1833) XL39 (reverse primer) (1929 .. 1948) BbsI (2046) EcoO109I (2053) BbvCI - Bpu10I (2149) BsmBI (2213) BstXI (2300) AgeI (2411) XcmI (2506) SexAI * (2598) SfiI (2784) StuI (2830) AvrII (2831) BclI * (2882) BsaBI * (2900) PstI (3094) pCMV6-AC-YFP 6631 bp
AanI  (6559)
2 sites
T T A T A A A A T A T T
DraIII  (6434)
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.
ScaI  (5762)
1 site
A G T A C T T C A T G A
AlwNI  (4808)
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  (4392)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (4013)
1 site
G T A T A C C A T A T G
BsmI  (3961)
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  (3900)
2 sites
T T A T A A A A T A T T
PfoI  (3816)
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.
BstBI  (3723)
1 site
T T C G A A A A G C T T
PflFI  (3159)
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  (3159)
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
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
AscI  (1030)
1 site
G G C G C G C C C C G C G C G G
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
BsiWI  (1070)
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.
NotI  (1076)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1082)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1082)
1 site
C T C G A G G A G C T C
BspEI  (1802)
1 site
T C C G G A A G G C C T
PmeI  (1823)
1 site
G T T T A A A C C A A A T T T G
FseI  (1833)
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.
BbsI  (2046)
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.
EcoO109I  (2053)
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.
BbvCI  (2149)
1 site
C C T C A G C G G A G T C G
Bpu10I  (2149)
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.
BsmBI  (2213)
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  (2300)
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  (2411)
1 site
A C C G G T T G G C C A
XcmI  (2506)
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  (2598)
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  (2784)
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  (2830)
1 site
A G G C C T T C C G G A
AvrII  (2831)
1 site
C C T A G G G G A T C C
BclI  (2882)
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  (2900)
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  (3094)
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
1929 .. 1948  =  20 annealed bases
Tm  =  59°C
AmpR
5209 .. 6069  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5209 .. 6000  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5209 .. 6069  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6001 .. 6069  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5209 .. 6069  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
2913 .. 3707  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2913 .. 3707  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
TurboYFP
1088 .. 1819  =  732 bp
243 amino acids  =  26.9 kDa
Product: enhanced derivative of PhiYFP
mammalian codon-optimized
TurboYFP
1088 .. 1819  =  732 bp
243 amino acids  =  26.9 kDa
Product: enhanced derivative of PhiYFP
mammalian codon-optimized
hGH poly(A) signal
1866 .. 2488  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
1866 .. 2488  =  623 bp
human growth hormone polyadenylation signal
ori
4453 .. 5038  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4453 .. 5038  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6201 .. 25  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6201 .. 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
2517 .. 2846  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
2517 .. 2846  =  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
3881 .. 4002  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3881 .. 4002  =  122 bp
SV40 polyadenylation signal
MCS
979 .. 1087  =  109 bp
multiple cloning sequence
MCS
979 .. 1087  =  109 bp
multiple cloning sequence
AmpR promoter
6070 .. 6174  =  105 bp
AmpR promoter
6070 .. 6174  =  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
2697 .. 2832  =  136 bp
SV40 origin of replication
SV40 ori
2697 .. 2832  =  136 bp
SV40 origin of replication
ORF:  3085 .. 3471  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  1088 .. 1819  =  732 bp
ORF:  243 amino acids  =  26.9 kDa
ORF:  5339 .. 5605  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2913 .. 3707  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1052 .. 1966  =  915 bp
ORF:  304 amino acids  =  30.4 kDa
ORF:  2230 .. 2454  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  5209 .. 6069  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1614 .. 1874  =  261 bp
ORF:  86 amino acids  =  9.7 kDa
ORF:  3222 .. 3758  =  537 bp
ORF:  178 amino acids  =  19.7 kDa
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