pPhi-Yellow-C

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

Sequence Author: Evrogen

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PciI (4649) EcoO109I (3829) PfoI (3506) RsrII (3247) BsrDI (2964) PflFI - Tth111I (2849) FspI (2833) EagI (2637) BspDI * - ClaI * (2571) StuI (2552) BseRI (2549) SfiI (2506) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (600) SgrAI (760) AleI (790) XmnI (850) PflMI (1014) BglII (1103) BlpI (1231) BssHII (1284) PaeR7I - XhoI (1319) Eco53kI (1324) SacI (1326) HindIII (1328) EcoRI (1335) PstI (1344) SalI (1345) AccI (1346) Acc65I (1351) KpnI (1355) PspOMI (1359) TspMI - XmaI (1362) ApaI (1363) SmaI (1364) BamHI (1366) XbaI * (1378) BclI * (1388) MfeI (1481) HpaI (1494) MluI (1617) DraIII (1847) CsiI - SexAI * (2320) pPhi-Yellow-C 4707 bp
PciI  (4649)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3829)
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.
PfoI  (3506)
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  (3247)
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  (2964)
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  (2849)
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  (2849)
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  (2833)
1 site
T G C G C A A C G C G T
EagI  (2637)
1 site
C G G C C G G C C G G C
BspDI  (2571)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2571)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2552)
1 site
A G G C C T T C C G G A
BseRI  (2549)
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.
SfiI  (2506)
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
AgeI  (600)
1 site
A C C G G T T G G C C A
SgrAI  (760)
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.
AleI  (790)
1 site
C A C N N N N G T G G T G N N N N C A C
XmnI  (850)
1 site
G A A N N N N T T C C T T N N N N A A G
PflMI  (1014)
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.
BglII  (1103)
1 site
A G A T C T T C T A G A
BlpI  (1231)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BssHII  (1284)
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.
PaeR7I  (1319)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1319)
1 site
C T C G A G G A G C T C
Eco53kI  (1324)
1 site
G A G C T C C T C G A G
SacI  (1326)
1 site
G A G C T C C T C G A G
HindIII  (1328)
1 site
A A G C T T T T C G A A
EcoRI  (1335)
1 site
G A A T T C C T T A A G
PstI  (1344)
1 site
C T G C A G G A C G T C
SalI  (1345)
1 site
G T C G A C C A G C T G
AccI  (1346)
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  (1351)
1 site
G G T A C C C C A T G G
KpnI  (1355)
1 site
G G T A C C C C A T G G
PspOMI  (1359)
1 site
G G G C C C C C C G G G
TspMI  (1362)
1 site
C C C G G G G G G C C C
XmaI  (1362)
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  (1363)
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  (1364)
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  (1366)
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  (1378)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1388)
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  (1481)
1 site
C A A T T G G T T A A C
HpaI  (1494)
1 site
G T T A A C C A A T T G
MluI  (1617)
1 site
A C G C G T T G C G C A
DraIII  (1847)
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  (2320)
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  (2320)
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
2603 .. 3397  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2603 .. 3397  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
PhiYFP
613 .. 1314  =  702 bp
234 amino acids  =  26.0 kDa
Product: Phialidium yellow fluorescent proteim
mammalian codon-optimized
PhiYFP
613 .. 1314  =  702 bp
234 amino acids  =  26.0 kDa
Product: Phialidium yellow fluorescent proteim
mammalian codon-optimized
ori
4005 .. 4593  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4005 .. 4593  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1623 .. 2078  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1623 .. 2078  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2211 .. 2568  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2211 .. 2568  =  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
1495 .. 1616  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1495 .. 1616  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2105 .. 2209  =  105 bp
AmpR promoter
2105 .. 2209  =  105 bp
MCS
1319 .. 1371  =  53 bp
multiple cloning site of fluorescent protein plasmids
MCS
1319 .. 1371  =  53 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3629 .. 3676  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3629 .. 3676  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2419 .. 2554  =  136 bp
SV40 origin of replication
SV40 ori
2419 .. 2554  =  136 bp
SV40 origin of replication
ORF:  613 .. 1386  =  774 bp
ORF:  257 amino acids  =  28.0 kDa
ORF:  3418 .. 3867  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2603 .. 3397  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2775 .. 3161  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  598 .. 1404  =  807 bp
ORF:  268 amino acids  =  26.7 kDa
ORF:  2912 .. 3448  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3623 .. 3856  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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