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Plasmid Files

pEX-K

Streamlined bacterial cloning vector with a kanamycin resistance marker.

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pEX-K.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Eurofins Genomics
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 PacI (2500) BmeT110I (2491) AvaI - BsoBI - PaeR7I - PspXI - XhoI (2490) HincII (2486) AccI (2485) SalI (2484) NdeI (2479) TatI (2462) pEX-For (2380 .. 2400) PfoI (2341) EcoO109I (2284) AatII (2230) ZraI (2228) AgeI (2104) RsrII (1953) NaeI (1939) NgoMIV (1937) MslI (1874) BtgI - NcoI (1869) SphI (1842) BssHII (1834) BanII (1802) BsaAI (1741) BsrDI (1670) NmeAIII (1661) PflFI - Tth111I (1555) FspI (1539) MscI (1519) PstI (1490) BmrI (1386) EagI (1343) BclI * (1278) BglII (1273) PacI (9) XbaI (13) NheI (19) BmtI (23) ApoI - EcoRI (25) pEX-Rev (75 .. 100) Bts α I (144) AflIII - PciI (358) AlwNI (774) BspEI (1170) XcmI (1253) pEX-K 2507 bp
PacI  (2500)
2 sites
T T A A T T A A A A T T A A T T
BmeT110I  (2491)
1 site
C Y C G R G G R G C Y C
AvaI  (2490)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2490)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
PaeR7I  (2490)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2490)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2490)
1 site
C T C G A G G A G C T C
HincII  (2486)
1 site
G T Y R A C C A R Y T G
AccI  (2485)
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.
SalI  (2484)
1 site
G T C G A C C A G C T G
NdeI  (2479)
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.
TatI  (2462)
1 site
W G T A C W W C A T G W
PfoI  (2341)
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.
EcoO109I  (2284)
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.
AatII  (2230)
1 site
G A C G T C C T G C A G
ZraI  (2228)
1 site
G A C G T C C T G C A G
AgeI  (2104)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
RsrII  (1953)
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.
NaeI  (1939)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
NgoMIV  (1937)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
MslI  (1874)
1 site
C A Y N N N N R T G G T R N N N N Y A C
BtgI  (1869)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1869)
1 site
C C A T G G G G T A C C
SphI  (1842)
1 site
G C A T G C C G T A C G
BssHII  (1834)
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.
BanII  (1802)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BsaAI  (1741)
1 site
Y A C G T R R T G C A Y
BsrDI  (1670)
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.
NmeAIII  (1661)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PflFI  (1555)
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  (1555)
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  (1539)
1 site
T G C G C A A C G C G T
MscI  (1519)
1 site
T G G C C A A C C G G T
PstI  (1490)
1 site
C T G C A G G A C G T C
BmrI  (1386)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
EagI  (1343)
1 site
C G G C C G G C C G G C
BclI  (1278)
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.
BglII  (1273)
1 site
A G A T C T T C T A G A
PacI  (9)
2 sites
T T A A T T A A A A T T A A T T
XbaI  (13)
1 site
T C T A G A A G A T C T
NheI  (19)
1 site
G C T A G C C G A T C G
BmtI  (23)
1 site
G C T A G C C G A T C G
ApoI  (25)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (25)
1 site
G A A T T C C T T A A G
BtsαI  (144)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
AflIII  (358)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (358)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (774)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
BspEI  (1170)
1 site
T C C G G A A G G C C T
XcmI  (1253)
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.
pEX-For
21-mer  /  67% GC
1 binding site
2380 .. 2400  =  21 annealed bases
Tm  =  63°C
forward sequencing primer
pEX-Rev
26-mer  /  50% GC
1 binding site
75 .. 100  =  26 annealed bases
Tm  =  62°C
reverse sequencing primer
NeoR/KanR
1309 .. 2103  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
1309 .. 2103  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
419 .. 1007  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
419 .. 1007  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
MCS
2478 .. 30  =  60 bp
multiple cloning site
MCS
2478 .. 30  =  60 bp
multiple cloning site
lac promoter
65 .. 95  =  31 bp
   Segment 3:  -10  
   65 .. 71  =  7 bp
promoter for the E. coli lac operon
lac promoter
65 .. 95  =  31 bp
   Segment 2:  
   72 .. 89  =  18 bp
promoter for the E. coli lac operon
lac promoter
65 .. 95  =  31 bp
   Segment 1:  -35  
   90 .. 95  =  6 bp
promoter for the E. coli lac operon
lac promoter
65 .. 95  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
41 .. 57  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
41 .. 57  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
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