pHcRed1-C1

Vector for fusing HcRed1 to the N-terminus of a partner protein.

Sequence Author: Clontech (TaKaRa)

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PciI (4640) BsaI (3711) PfoI (3497) RsrII (3238) BsrDI (2955) PflFI - Tth111I (2840) MscI (2804) BspDI * - ClaI * (2562) StuI (2543) BseRI (2540) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (600) EcoNI (628) FspAI (643) BsrGI (653) PmlI (747) PpuMI (815) XcmI (1047) BssHII (1164) ScaI (1237) BspEI (1297) BglII (1306) PaeR7I - XhoI (1310) Eco53kI (1315) SacI (1317) HindIII (1319) EcoRI (1326) PstI (1335) SalI (1336) AccI (1337) Acc65I (1342) KpnI (1346) SacII (1349) PspOMI (1350) TspMI - XmaI (1353) ApaI (1354) SmaI (1355) BamHI (1357) XbaI * (1369) stop codons BclI * (1379) MfeI (1472) HpaI (1485) BtsI - BtsαI (1561) MluI (1608) DraIII (1838) CsiI - SexAI * (2311) pHcRed1-C1 4698 bp
PciI  (4640)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BsaI  (3711)
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  (3497)
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  (3238)
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  (2955)
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  (2840)
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  (2840)
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.
MscI  (2804)
1 site
T G G C C A A C C G G T
BspDI  (2562)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2562)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2543)
1 site
A G G C C T T C C G G A
BseRI  (2540)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
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
AgeI  (600)
1 site
A C C G G T T G G C C A
EcoNI  (628)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
FspAI  (643)
1 site
R T G C G C A Y Y A C G C G T R
BsrGI  (653)
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.
PmlI  (747)
1 site
C A C G T G G T G C A C
PpuMI  (815)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
XcmI  (1047)
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.
BssHII  (1164)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
ScaI  (1237)
1 site
A G T A C T T C A T G A
BspEI  (1297)
1 site
T C C G G A A G G C C T
BglII  (1306)
1 site
A G A T C T T C T A G A
PaeR7I  (1310)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1310)
1 site
C T C G A G G A G C T C
Eco53kI  (1315)
1 site
G A G C T C C T C G A G
SacI  (1317)
1 site
G A G C T C C T C G A G
HindIII  (1319)
1 site
A A G C T T T T C G A A
EcoRI  (1326)
1 site
G A A T T C C T T A A G
PstI  (1335)
1 site
C T G C A G G A C G T C
SalI  (1336)
1 site
G T C G A C C A G C T G
AccI  (1337)
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  (1342)
1 site
G G T A C C C C A T G G
KpnI  (1346)
1 site
G G T A C C C C A T G G
SacII  (1349)
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  (1350)
1 site
G G G C C C C C C G G G
TspMI  (1353)
1 site
C C C G G G G G G C C C
XmaI  (1353)
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  (1354)
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  (1355)
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  (1357)
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.
XbaI  (1369)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1379)
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.
MfeI  (1472)
1 site
C A A T T G G T T A A C
HpaI  (1485)
1 site
G T T A A C C A A T T G
BtsI  (1561)
1 site
G C A G T G N N C G T C A C
BtsαI  (1561)
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.
MluI  (1608)
1 site
A C G C G T T G C G C A
DraIII  (1838)
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  (2311)
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  (2311)
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
2594 .. 3388  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2594 .. 3388  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
HcRed1
613 .. 1296  =  684 bp
228 amino acids  =  25.8 kDa
Product: far-red fluorescent variant of Heteractis crispa chromoprotein
mammalian codon-optimized
HcRed1
613 .. 1296  =  684 bp
228 amino acids  =  25.8 kDa
Product: far-red fluorescent variant of Heteractis crispa chromoprotein
mammalian codon-optimized
ori
3996 .. 4584  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3996 .. 4584  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV promoter
52 .. 568  =  517 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
52 .. 568  =  517 bp
human cytomegalovirus (CMV) immediate early promoter
f1 ori
1614 .. 2069  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1614 .. 2069  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2202 .. 2559  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2202 .. 2559  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
1486 .. 1607  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1486 .. 1607  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2096 .. 2200  =  105 bp
AmpR promoter
2096 .. 2200  =  105 bp
MCS
1297 .. 1362  =  66 bp
multiple cloning site of fluorescent protein plasmids
MCS
1297 .. 1362  =  66 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3620 .. 3667  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3620 .. 3667  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
stop codons
1371 .. 1381  =  11 bp
stop codons in all three reading frames
stop codons
1371 .. 1381  =  11 bp
stop codons in all three reading frames
SV40 ori
2410 .. 2545  =  136 bp
SV40 origin of replication
SV40 ori
2410 .. 2545  =  136 bp
SV40 origin of replication
ORF:  613 .. 1377  =  765 bp
ORF:  254 amino acids  =  28.2 kDa
ORF:  3409 .. 3858  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2594 .. 3388  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2766 .. 3152  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  598 .. 1395  =  798 bp
ORF:  265 amino acids  =  26.0 kDa
ORF:  2903 .. 3439  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3614 .. 3847  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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