pPAmCherry1-N1

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

Sequence Author: Verkhusha Lab

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PciI (4665) BsaI (3736) PfoI (3522) RsrII (3263) BsrDI (2980) PflFI - Tth111I (2865) FspI (2849) BspDI * - ClaI * (2587) StuI (2568) SfiI (2522) 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) PmlI (768) SbfI (1034) BbsI (1114) PflMI (1118) BbvCI (1217) BsgI (1273) SgrAI (1360) XcmI (1365) BsrGI (1379) NotI (1391) XbaI * (1401) MfeI (1497) HpaI (1510) BtsI - BtsαI (1586) AflII (1629) DraIII (1863) CsiI - SexAI * (2336) pPAmCherry1-N1 4723 bp
PciI  (4665)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BsaI  (3736)
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  (3522)
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  (3263)
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  (2980)
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  (2865)
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  (2865)
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  (2849)
1 site
T G C G C A A C G C G T
BspDI  (2587)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2587)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2568)
1 site
A G G C C T T C C G G A
SfiI  (2522)
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

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
PmlI  (768)
1 site
C A C G T G G T G C A C
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
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  (1391)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1401)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1497)
1 site
C A A T T G G T T A A C
HpaI  (1510)
1 site
G T T A A C C A A T T G
BtsI  (1586)
1 site
G C A G T G N N C G T C A C
BtsαI  (1586)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
AflII  (1629)
1 site
C T T A A G G A A T T C
DraIII  (1863)
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.
CsiI  (2336)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (2336)
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
2619 .. 3413  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2619 .. 3413  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
PAmCherry1
679 .. 1389  =  711 bp
236 amino acids  =  26.8 kDa
Product: photoactivatable monomeric derivative of DsRed fluorescent protein
mammalian codon-optimized
PAmCherry1
679 .. 1389  =  711 bp
236 amino acids  =  26.8 kDa
Product: photoactivatable monomeric derivative of DsRed fluorescent protein
mammalian codon-optimized
ori
4021 .. 4609  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4021 .. 4609  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1639 .. 2094  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1639 .. 2094  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2227 .. 2584  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2227 .. 2584  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
60 .. 364  =  305 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
60 .. 364  =  305 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
1511 .. 1632  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1511 .. 1632  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2121 .. 2225  =  105 bp
AmpR promoter
2121 .. 2225  =  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
3645 .. 3692  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3645 .. 3692  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2435 .. 2570  =  136 bp
SV40 origin of replication
SV40 ori
2435 .. 2570  =  136 bp
SV40 origin of replication
ORF:  679 .. 1389  =  711 bp
ORF:  236 amino acids  =  26.8 kDa
ORF:  2791 .. 3177  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3434 .. 3883  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2619 .. 3413  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  447 .. 746  =  300 bp
ORF:  99 amino acids  =  10.6 kDa
ORF:  2928 .. 3464  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3639 .. 3872  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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