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Plasmid Files

pmCherry-N1

Vector for fusing mCherry to the C-terminus of a partner protein.

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pmCherry-N1 Sequence and MappmCherry-N1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 AflIII - PciI (4664) EcoO109I (3844) BsaI (3735) PfoI (3521) RsrII (3262) BsrDI (2979) PflFI - Tth111I (2864) FspI (2848) BspDI * - ClaI * (2586) SfiI (2521) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) BglII (609) PaeR7I - XhoI (613) Eco53kI (618) SacI (620) HindIII (622) EcoRI (629) SalI (639) AccI (640) Acc65I (645) KpnI (649) SacII (652) PspOMI (653) TspMI - XmaI (656) ApaI (657) SmaI (658) BamHI (660) AgeI (666) AhdI (873) SbfI (1034) BbsI (1114) PflMI (1118) BbvCI - Bpu10I (1217) BsgI (1273) SgrAI (1360) XcmI (1365) BsrGI (1379) NotI (1390) XbaI * (1400) MfeI (1496) HpaI (1509) Bts α I (1585) AflII (1628) DraIII (1862) SexAI * (2335) pmCherry-N1 4722 bp
AflIII  (4664)
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  (4664)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3844)
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.
BsaI  (3735)
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.
PfoI  (3521)
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.
RsrII  (3262)
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.
BsrDI  (2979)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PflFI  (2864)
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  (2864)
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.
FspI  (2848)
1 site
T G C G C A A C G C G T
BspDI  (2586)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2586)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SfiI  (2521)
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.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
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  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
AfeI  (596)
1 site
A G C G C T T C G C G A
BglII  (609)
1 site
A G A T C T T C T A G A
PaeR7I  (613)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (613)
1 site
C T C G A G G A G C T C
Eco53kI  (618)
1 site
G A G C T C C T C G A G
SacI  (620)
1 site
G A G C T C C T C G A G
HindIII  (622)
1 site
A A G C T T T T C G A A
EcoRI  (629)
1 site
G A A T T C C T T A A G
SalI  (639)
1 site
G T C G A C C A G C T G
AccI  (640)
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.
Acc65I  (645)
1 site
G G T A C C C C A T G G
KpnI  (649)
1 site
G G T A C C C C A T G G
SacII  (652)
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.
PspOMI  (653)
1 site
G G G C C C C C C G G G
TspMI  (656)
1 site
C C C G G G G G G C C C
XmaI  (656)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
ApaI  (657)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SmaI  (658)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (660)
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.
AgeI  (666)
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.
AhdI  (873)
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.
SbfI  (1034)
1 site
C C T G C A G G G G A C G T C C
BbsI  (1114)
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.
PflMI  (1118)
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.
BbvCI  (1217)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1217)
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.
BsgI  (1273)
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).
SgrAI  (1360)
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.
XcmI  (1365)
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.
BsrGI  (1379)
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.
NotI  (1390)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1400)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1496)
1 site
C A A T T G G T T A A C
HpaI  (1509)
1 site
G T T A A C C A A T T G
BtsαI  (1585)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
AflII  (1628)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
DraIII  (1862)
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.
SexAI  (2335)
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.
NeoR/KanR
2618 .. 3412  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
2618 .. 3412  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
mCherry
679 .. 1389  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
mCherry
679 .. 1389  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
ori
4020 .. 4608  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4020 .. 4608  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1638 .. 2093  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1638 .. 2093  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
2226 .. 2583  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2226 .. 2583  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
1510 .. 1631  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1510 .. 1631  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2120 .. 2224  =  105 bp
AmpR promoter
2120 .. 2224  =  105 bp
MCS
591 .. 671  =  81 bp
multiple cloning site of fluorescent protein plasmids
MCS
591 .. 671  =  81 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3644 .. 3691  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3644 .. 3691  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2434 .. 2569  =  136 bp
SV40 origin of replication
SV40 ori
2434 .. 2569  =  136 bp
SV40 origin of replication
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