pCMV-Myc-C

Mammalian vector for CMV promoter-driven expression of an ORF with a C-terminal Myc tag.

Sequence Author: transOMIC, Clontech (TaKaRa)

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PluTI (3616) SfoI (3614) NarI (3613) KasI (3612) BstAPI (3562) SspI (3194) XmnI (2989) ScaI (2870) NmeAIII (2538) BsaI (2451) AhdI (2390) PspFI (1805) BfuAI - BspMI (12) SbfI (23) BtgZI (300) BsaAI - SnaBI (306) NcoI - StyI (326) Eco53kI (532) SacI (534) BbsI (600) BspEI (617) StuI (769) PspOMI * (820) ApaI * (824) EcoRI (832) SalI (839) AccI (840) BglII (847) MCS AvaI - BsoBI - PaeR7I - XhoI (853) Acc65I (858) KpnI (862) EagI - NotI (901) PsiI (1025) MfeI (1054) NheI (1136) BmtI (1140) M13 rev BspQI - SapI (1381) AflIII - PciI (1497) BseYI (1801) pCMV-Myc-C 3772 bp
PluTI  (3616)
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  (3614)
1 site
G G C G C C C C G C G G
NarI  (3613)
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  (3612)
1 site
G G C G C C C C G C G G
BstAPI  (3562)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
SspI  (3194)
1 site
A A T A T T T T A T A A
XmnI  (2989)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2870)
1 site
A G T A C T T C A T G A
NmeAIII  (2538)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsaI  (2451)
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.
AhdI  (2390)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
PspFI  (1805)
1 site
C C C A G C G G G T C G
BfuAI  (12)
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  (12)
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.
SbfI  (23)
1 site
C C T G C A G G G G A C G T C C
BtgZI  (300)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BsaAI  (306)
1 site
Y A C G T R R T G C A Y
SnaBI  (306)
1 site
T A C G T A A T G C A T
NcoI  (326)
1 site
C C A T G G G G T A C C
StyI  (326)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
Eco53kI  (532)
1 site
G A G C T C C T C G A G
SacI  (534)
1 site
G A G C T C C T C G A G
BbsI  (600)
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.
BspEI  (617)
1 site
T C C G G A A G G C C T
StuI  (769)
1 site
A G G C C T T C C G G A
PspOMI  (820)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI  (824)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI can be used between 25°C and 37°C.
EcoRI  (832)
1 site
G A A T T C C T T A A G
SalI  (839)
1 site
G T C G A C C A G C T G
AccI  (840)
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.
BglII  (847)
1 site
A G A T C T T C T A G A
AvaI  (853)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (853)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (853)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (853)
1 site
C T C G A G G A G C T C
Acc65I  (858)
1 site
G G T A C C C C A T G G
KpnI  (862)
1 site
G G T A C C C C A T G G
EagI  (901)
1 site
C G G C C G G C C G G C
NotI  (901)
1 site
G C G G C C G C C G C C G G C G
PsiI  (1025)
1 site
T T A T A A A A T A T T
MfeI  (1054)
1 site
C A A T T G G T T A A C
NheI  (1136)
1 site
G C T A G C C G A T C G
BmtI  (1140)
1 site
G C T A G C C G A T C G
BspQI  (1381)
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  (1381)
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.
AflIII  (1497)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (1497)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (1801)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AmpR
2317 .. 3177  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2317 .. 3108  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2317 .. 3177  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3109 .. 3177  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2317 .. 3177  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1558 .. 2146  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
1558 .. 2146  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
27 .. 330  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
27 .. 330  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
331 .. 534  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
331 .. 534  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
911 .. 1045  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
911 .. 1045  =  135 bp
SV40 polyadenylation signal
AmpR promoter
3178 .. 3282  =  105 bp
AmpR promoter
3178 .. 3282  =  105 bp
SV40 intron
672 .. 768  =  97 bp
modified SV40 intron with splice donor and acceptor sites
SV40 intron
672 .. 768  =  97 bp
modified SV40 intron with splice donor and acceptor sites
MCS
832 .. 863  =  32 bp
multiple cloning site
MCS
832 .. 863  =  32 bp
multiple cloning site
Myc
867 .. 896  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
867 .. 896  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
M13 rev
1156 .. 1172  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
1156 .. 1172  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  2447 .. 2713  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3563 .. 36  =  246 bp
ORF:  81 amino acids  =  9.5 kDa
ORF:  784 .. 1062  =  279 bp
ORF:  92 amino acids  =  10.2 kDa
ORF:  2317 .. 3177  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3523 .. 113  =  363 bp
ORF:  120 amino acids  =  13.4 kDa
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