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Plasmid Files

pmKate2-C

Vector for fusing mKate2 to the N-terminus of a partner protein.

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pmKate2-C.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Evrogen
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PciI (4664) EcoO109I (3844) RsrII (3262) BsrDI (2979) PflFI - Tth111I (2864) FspI (2848) PluTI (2749) SfoI (2747) NarI (2746) KasI (2745) EagI (2652) BspDI * - ClaI * (2586) SfiI (2521) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (600) BsrGI (653) PshAI (1243) BspEI (1321) BglII (1330) PaeR7I - XhoI (1334) Eco53kI (1339) SacI (1341) HindIII (1343) EcoRI (1350) PstI (1359) SalI (1360) Acc65I (1366) KpnI (1370) SacII (1373) PspOMI (1374) TspMI - XmaI (1377) ApaI (1378) SmaI (1379) BamHI (1381) XbaI * (1393) BclI * (1403) MfeI (1496) HpaI (1509) Bts α I (1585) MluI (1632) SexAI * (2335) pmKate2-C 4722 bp
PciI  (4664)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3844)
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.
RsrII  (3262)
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  (2979)
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  (2864)
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  (2864)
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  (2848)
1 site
T G C G C A A C G C G T
PluTI  (2749)
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  (2747)
1 site
G G C G C C C C G C G G
NarI  (2746)
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  (2745)
1 site
G G C G C C C C G C G G
EagI  (2652)
1 site
C G G C C G G C C G G C
BspDI  (2586)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2586)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SfiI  (2521)
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
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.
PshAI  (1243)
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  (1321)
1 site
T C C G G A A G G C C T
BglII  (1330)
1 site
A G A T C T T C T A G A
PaeR7I  (1334)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1334)
1 site
C T C G A G G A G C T C
Eco53kI  (1339)
1 site
G A G C T C C T C G A G
SacI  (1341)
1 site
G A G C T C C T C G A G
HindIII  (1343)
1 site
A A G C T T T T C G A A
EcoRI  (1350)
1 site
G A A T T C C T T A A G
PstI  (1359)
1 site
C T G C A G G A C G T C
SalI  (1360)
1 site
G T C G A C C A G C T G
Acc65I  (1366)
1 site
G G T A C C C C A T G G
KpnI  (1370)
1 site
G G T A C C C C A T G G
SacII  (1373)
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  (1374)
1 site
G G G C C C C C C G G G
TspMI  (1377)
1 site
C C C G G G G G G C C C
XmaI  (1377)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
ApaI  (1378)
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  (1379)
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  (1381)
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  (1393)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1403)
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  (1496)
1 site
C A A T T G G T T A A C
HpaI  (1509)
1 site
G T T A A C C A A T T G
BtsαI  (1585)
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  (1632)
1 site
A C G C G T T G C G C A
SexAI  (2335)
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
2618 .. 3412  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2618 .. 3412  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
mKate2
613 .. 1308  =  696 bp
232 amino acids  =  26.1 kDa
Product: monomeric far-red fluorescent protein
mammalian codon-optimized
mKate2
613 .. 1308  =  696 bp
232 amino acids  =  26.1 kDa
Product: monomeric far-red fluorescent protein
mammalian codon-optimized
ori
4020 .. 4608  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4020 .. 4608  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1638 .. 2093  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1638 .. 2093  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2226 .. 2583  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2226 .. 2583  =  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
1510 .. 1631  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1510 .. 1631  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2120 .. 2224  =  105 bp
AmpR promoter
2120 .. 2224  =  105 bp
MCS
1321 .. 1386  =  66 bp
multiple cloning site of fluorescent protein plasmids
MCS
1321 .. 1386  =  66 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3644 .. 3691  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3644 .. 3691  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2434 .. 2569  =  136 bp
SV40 origin of replication
SV40 ori
2434 .. 2569  =  136 bp
SV40 origin of replication
ORF:  613 .. 1401  =  789 bp
ORF:  262 amino acids  =  28.7 kDa
ORF:  3433 .. 3882  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2618 .. 3412  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2790 .. 3176  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  598 .. 1419  =  822 bp
ORF:  273 amino acids  =  28.0 kDa
ORF:  717 .. 962  =  246 bp
ORF:  81 amino acids  =  9.1 kDa
ORF:  2927 .. 3463  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3638 .. 3871  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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