pME-HA (linearized)

Linearized mammalian cell expression vector for directional recombination-based cloning of a gene in the Expresso® system.

Sequence Author: Lucigen

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Eco53kI (3324) CMV promoter SnaBI (3098) ApaI (2755) PspOMI (2751) ApaLI (2439) XcmI (2034) BsaI (1911) PfoI (1697) BstBI (1604) SacI (3326) pME Forward (3344 .. 3365) BtsI - BtsαI (3346) NheI (3382) BmtI (3386) BmgBI (12) HA AvaI - BsoBI - PaeR7I - XhoI (38) pME Reverse (59 .. 80) PsiI (208) ApoI (263) AseI (277) XmnI (278) BspHI (348) SspI (383) Bsu36I (402) CsiI - SexAI * (511) SfiI (697) BseRI (740) StuI (743) BsaBI * (781) EagI (828) KasI (921) NarI (922) SfoI (923) PluTI (925) DrdI (949) MscI (1004) FspI (1024) PflFI - Tth111I (1040) BsrDI (1155) BtgI (1354) RsrII (1438) pME-HA 3416 bp
Eco53kI  (3324)
1 site
G A G C T C C T C G A G
SnaBI  (3098)
1 site
T A C G T A A T G C A T
ApaI  (2755)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (2751)
1 site
G G G C C C C C C G G G
ApaLI  (2439)
1 site
G T G C A C C A C G T G
XcmI  (2034)
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.
BsaI  (1911)
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  (1697)
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.
BstBI  (1604)
1 site
T T C G A A A A G C T T
SacI  (3326)
1 site
G A G C T C C T C G A G
BtsI  (3346)
1 site
G C A G T G N N C G T C A C
BtsαI  (3346)
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.
NheI  (3382)
1 site
G C T A G C C G A T C G
BmtI  (3386)
1 site
G C T A G C C G A T C G
BmgBI  (12)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
AvaI  (38)
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  (38)
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  (38)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (38)
1 site
C T C G A G G A G C T C
PsiI  (208)
1 site
T T A T A A A A T A T T
ApoI  (263)
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.
AseI  (277)
1 site
A T T A A T T A A T T