pLSSmOrange-C1

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

Sequence Author: Verkhusha Lab

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PciI (4658) ApaLI (4344) EcoO109I (3838) BsaI (3729) PfoI (3515) RsrII (3256) BsrDI (2973) PflFI - Tth111I (2858) EagI (2646) BspDI * - ClaI * (2580) SfiI (2515) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) FspAI (670) AhdI (801) SbfI (962) BbsI (1042) BstXI (1047) Bpu10I (1108) BsgI (1201) SgrAI (1288) XcmI (1293) BsrGI (1307) BspEI (1315) BglII (1324) PaeR7I - XhoI (1328) Eco53kI (1333) SacI (1335) HindIII (1337) EcoRI (1344) Acc65I (1360) KpnI (1364) SacII (1367) PspOMI (1368) TspMI - XmaI (1371) ApaI (1372) SmaI (1373) BamHI (1375) XbaI * (1387) stop codons BclI * (1397) MfeI (1490) HpaI (1503) BtsI - BtsαI (1579) MluI (1626) DraIII (1856) CsiI - SexAI * (2329) pLSSmOrange-C1 4716 bp
PciI  (4658)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (4344)
1 site
G T G C A C C A C G T G
EcoO109I  (3838)
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  (3729)
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  (3515)
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  (3256)
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  (2973)
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  (2858)
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  (2858)
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.
EagI  (2646)
1 site
C G G C C G G C C G G C
BspDI  (2580)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2580)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SfiI  (2515)
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
FspAI  (670)
1 site
R T G C G C A Y Y A C G C G T R
AhdI  (801)
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  (962)
1 site
C C T G C A G G G G A C G T C C
BbsI  (1042)
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.
BstXI  (1047)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
Bpu10I  (1108)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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  (1201)
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  (1288)
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  (1293)
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  (1307)
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.
BspEI  (1315)
1 site
T C C G G A A G G C C T
BglII  (1324)
1 site
A G A T C T T C T A G A
PaeR7I  (1328)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1328)
1 site
C T C G A G G A G C T C
Eco53kI  (1333)
1 site
G A G C T C C T C G A G
SacI  (1335)
1 site
G A G C T C C T C G A G
HindIII  (1337)
1 site
A A G C T T T T C G A A
EcoRI  (1344)
1 site
G A A T T C C T T A A G
Acc65I  (1360)
1 site
G G T A C C C C A T G G
KpnI  (1364)
1 site
G G T A C C C C A T G G
SacII  (1367)
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  (1368)
1 site
G G G C C C C C C G G G
TspMI  (1371)
1 site
C C C G G G G G G C C C
XmaI  (1371)
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  (1372)
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  (1373)
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  (1375)
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  (1387)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1397)
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  (1490)
1 site
C A A T T G G T T A A C
HpaI  (1503)
1 site
G T T A A C C A A T T G
BtsI  (1579)
1 site
G C A G T G N N C G T C A C
BtsαI  (1579)
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  (1626)
1 site
A C G C G T T G C G C A
DraIII  (1856)
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  (2329)
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  (2329)
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
2612 .. 3406  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2612 .. 3406  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
LSSmOrange
607 .. 1314  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric orange fluorescent protein with a large Stokes shift
mammalian codon-optimized
LSSmOrange
607 .. 1314  =  708 bp
236 amino acids  =  26.7 kDa
Product: monomeric orange fluorescent protein with a large Stokes shift
mammalian codon-optimized
ori
4014 .. 4602  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4014 .. 4602  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1632 .. 2087  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1632 .. 2087  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2220 .. 2577  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2220 .. 2577  =  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
1504 .. 1625  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1504 .. 1625  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2114 .. 2218  =  105 bp