pDream2.1 MCS

Vector for expression of N-terminally FLAG-tagged proteins in bacterial, insect, and mammalian cells.

Sequence Author: GenScript

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SalI (243) ScaI (6790) AlwNI (5991) PciI (5576) AsiSI (5432) BmgBI (5390) baculovirus recombination region (ORF1629) SwaI (4976) AfeI (4844) β-globin poly(A) signal BspHI (4486) BsmI (4434) BstBI (4196) RsrII (4030) BssHII (3911) PflFI - Tth111I (3632) PstI (3567) PluTI (3517) SfoI (3515) AccI (244) HincII (245) MluI (399) baculovirus recombination region (lef2/ORF603) FseI (1017) PaeR7I - PspXI - XhoI (1020) T7 primer (1615 .. 1634) T7 promoter lac operator PpuMI (1660) PacI (1732) attB1 ATG FLAG BamHI (1837) BglII (1844) EcoRI (1851) Acc65I (1857) KpnI (1861) Eco53kI (1865) SacI (1867) HindIII (1869) NotI (1876) SP6 primer (1883 .. 1904) attB2 AleI (1988) hGH poly(A) signal BsgI (2172) BlpI (2258) T7 terminator XbaI (2456) DraIII (2781) CsiI - SexAI * (3071) BseRI (3300) StuI (3303) AvrII (3304) TspMI - XmaI (3325) SmaI (3327) BclI * (3355) BsaBI * (3373) KasI (3513) NarI (3514) pDream2.1/MCS 7159 bp
SalI  (243)
1 site
G T C G A C C A G C T G
ScaI  (6790)
1 site
A G T A C T T C A T G A
AlwNI  (5991)
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.
PciI  (5576)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AsiSI  (5432)
1 site
G C G A T C G C C G C T A G C G
BmgBI  (5390)
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.
SwaI  (4976)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
AfeI  (4844)
1 site
A G C G C T T C G C G A
BspHI  (4486)
1 site
T C A T G A A G T A C T
BsmI  (4434)
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.
BstBI  (4196)
1 site
T T C G A A A A G C T T
RsrII  (4030)
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  (3911)
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.
PflFI  (3632)
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  (3632)
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.
PstI  (3567)
1 site
C T G C A G G A C G T C
PluTI  (3517)
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  (3515)
1 site
G G C G C C C C G C G G
AccI  (244)
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.
HincII  (245)
1 site
G T Y R A C C A R Y T G
MluI  (399)
1 site
A C G C G T T G C G C A
FseI  (1017)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
PaeR7I  (1020)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1020)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1020)
1 site
C T C G A G G A G C T C
PpuMI  (1660)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
PacI  (1732)
1 site
T T A A T T A A A A T T A A T T
BamHI  (1837)
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.
BglII  (1844)
1 site
A G A T C T T C T A G A
EcoRI  (1851)
1 site
G A A T T C C T T A A G
Acc65I  (1857)
1 site
G G T A C C C C A T G G
KpnI  (1861)
1 site
G G T A C C C C A T G G
Eco53kI  (1865)
1 site
G A G C T C C T C G A G
SacI  (1867)
1 site
G A G C T C C T C G A G
HindIII  (1869)
1 site
A A G C T T T T C G A A
NotI  (1876)
1 site
G C G G C C G C C G C C G G C G
AleI  (1988)
1 site
C A C N N N N G T G G T G N N N N C A C
BsgI  (2172)
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).
BlpI  (2258)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
XbaI  (2456)
1 site
T C T A G