pKatushka2S-C

Vector for fusing Katushka2S to the N-terminus of a partner protein.

Sequence Author: Evrogen

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PciI (4661) EcoO109I (3841) PfoI (3518) RsrII (3259) BsrDI (2976) PflFI - Tth111I (2861) FspI (2845) PluTI (2746) SfoI (2744) NarI (2743) KasI (2742) EagI (2649) BspDI * - ClaI * (2583) StuI (2564) BseRI (2561) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (600) AleI (613) BstXI (615) BsrGI (662) BbsI (920) BsgI (1117) PshAI (1252) BspEI (1318) BglII (1327) PaeR7I - XhoI (1331) Eco53kI (1336) SacI (1338) HindIII (1340) EcoRI (1347) PstI (1356) SalI (1357) AccI (1358) Acc65I (1363) KpnI (1367) SacII (1370) PspOMI (1371) TspMI - XmaI (1374) ApaI (1375) SmaI (1376) BamHI (1378) XbaI * (1390) MfeI (1493) HpaI (1506) MluI (1629) DraIII (1859) CsiI - SexAI * (2332) pKatushka2S-C 4719 bp
PciI  (4661)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3841)
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  (3518)
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  (3259)
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  (2976)
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  (2861)
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  (2861)
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  (2845)
1 site
T G C G C A A C G C G T
PluTI  (2746)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (2744)
1 site
G G C G C C C C G C G G
NarI  (2743)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (2742)
1 site
G G C G C C C C G C G G
EagI  (2649)
1 site
C G G C C G G C C G G C
BspDI  (2583)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2583)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2564)
1 site
A G G C C T T C C G G A
BseRI  (2561)
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
AleI  (613)
1 site
C A C N N N N G T G G T G N N N N C A C
BstXI  (615)
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.
BsrGI  (662)
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.
BbsI  (920)
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.
BsgI  (1117)
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).
PshAI  (1252)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BspEI  (1318)
1 site
T C C G G A A G G C C T
BglII  (1327)
1 site
A G A T C T T C T A G A
PaeR7I  (1331)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1331)
1 site
C T C G A G G A G C T C
Eco53kI  (1336)
1 site
G A G C T C C T C G A G
SacI  (1338)
1 site
G A G C T C C T C G A G
HindIII  (1340)
1 site
A A G C T T T T C G A A
EcoRI  (1347)
1 site
G A A T T C C T T A A G
PstI  (1356)
1 site
C T G C A G G A C G T C
SalI  (1357)
1 site
G T C G A C C A G C T G
AccI  (1358)
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  (1363)
1 site
G G T A C C C C A T G G
KpnI  (1367)
1 site
G G T A C C C C A T G G
SacII  (1370)
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  (1371)
1 site
G G G C C C C C C G G G
TspMI  (1374)
1 site
C C C G G G G G G C C C
XmaI  (1374)
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  (1375)
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  (1376)
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  (1378)
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  (1390)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1493)
1 site
C A A T T G G T T A A C
HpaI  (1506)
1 site
G T T A A C C A A T T G
MluI  (1629)
1 site
A C G C G T T G C G C A
DraIII  (1859)
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  (2332)
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  (2332)
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
2615 .. 3409  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2615 .. 3409  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
Katushka2S
613 .. 1317  =  705 bp
235 amino acids  =  26.4 kDa
Product: improved version of the oligomeric far-red fluorescent protein TurboFP635/Katushka (Luker et al., 2015)
mammalian codon-optimized
Katushka2S
613 .. 1317  =  705 bp
235 amino acids  =  26.4 kDa
Product: improved version of the oligomeric far-red fluorescent protein TurboFP635/Katushka (Luker et al., 2015)
mammalian codon-optimized
ori
4017 .. 4605  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4017 .. 4605  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1635 .. 2090  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1635 .. 2090  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2223 .. 2580  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2223 .. 2580  =  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
1507 .. 1628  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1507 .. 1628  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2117 .. 2221  =  105 bp
AmpR promoter
2117 .. 2221  =  105 bp
MCS
1318 .. 1383  =  66 bp
multiple cloning site
MCS
1318 .. 1383  =  66 bp
multiple cloning site
HSV TK poly(A) signal
3641 .. 3688  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
3641 .. 3688  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
SV40 ori
2431 .. 2566  =  136 bp
SV40 origin of replication
SV40 ori
2431 .. 2566  =  136 bp
SV40 origin of replication
ORF:  613 .. 1398  =  786 bp
ORF:  261 amino acids  =  28.8 kDa
ORF:  3430 .. 3879  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2615 .. 3409  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2787 .. 3173  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  598 .. 1416  =  819 bp
ORF:  272 amino acids  =  28.1 kDa
ORF:  726 .. 971  =  246 bp
ORF:  81 amino acids  =  9.2 kDa
ORF:  2924 .. 3460  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3635 .. 3868  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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