pGen2.1

CMV promoter-based mammalian expression vector encoding an N-terminal FLAG tag.

Sequence Author: GenScript

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SgrDI (5772) ScaI (5331) PvuI (5221) PciI (3961) BstZ17I (3582) BsmI (3530) PfoI (3385) BstBI (3292) RsrII (3126) BssHII (3007) BglII (12) MfeI (161) NruI (208) MluI (228) SpeI (249) NdeI (484) SnaBI (590) T7 primer (863 .. 882) NheI (895) BmtI (899) PmeI (904) AflII (908) HindIII (911) ATG FLAG NotI (976) XbaI (994) BamHI (1000) PspOMI (1031) ApaI (1035) SP6 promoter AleI (1136) Reverse sequencing primer (1173 .. 1192) BsgI (1320) BbvCI (1362) BlpI (1406) BsmBI - Esp3I (1426) BstXI (1513) BbsI (1563) DraIII (1877) CsiI - SexAI * (2167) BseRI (2396) StuI (2399) AvrII (2400) TspMI - XmaI (2421) SmaI (2423) BsaBI * (2469) KasI (2609) NarI (2610) SfoI (2611) PluTI (2613) PstI (2663) PflFI - Tth111I (2728) pGen2.1 5774 bp
SgrDI  (5772)
1 site
C G T C G A C G G C A G C T G C
ScaI  (5331)
1 site
A G T A C T T C A T G A
PvuI  (5221)
1 site
C G A T C G G C T A G C
PciI  (3961)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstZ17I  (3582)
1 site
G T A T A C C A T A T G
BsmI  (3530)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
PfoI  (3385)
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  (3292)
1 site
T T C G A A A A G C T T
RsrII  (3126)
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.
BssHII  (3007)
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.
BglII  (12)
1 site
A G A T C T T C T A G A
MfeI  (161)
1 site
C A A T T G G T T A A C
NruI  (208)
1 site
T C G C G A A G C G C T
MluI  (228)
1 site
A C G C G T T G C G C A
SpeI  (249)
1 site
A C T A G T T G A T C A
NdeI  (484)
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  (590)
1 site
T A C G T A A T G C A T
NheI  (895)
1 site
G C T A G C C G A T C G
BmtI  (899)
1 site
G C T A G C C G A T C G
PmeI  (904)
1 site
G T T T A A A C C A A A T T T G
AflII  (908)
1 site
C T T A A G G A A T T C
HindIII  (911)
1 site
A A G C T T T T C G A A
NotI  (976)
1 site
G C G G C C G C C G C C G G C G
XbaI  (994)
1 site
T C T A G A A G A T C T
BamHI  (1000)
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.
PspOMI  (1031)
1 site
G G G C C C C C C G G G
ApaI  (1035)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AleI  (1136)
1 site
C A C N N N N G T G G T G N N N N C A C
BsgI  (1320)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14