pFLD(alpha)

Pichia pastoris vector for methylamine-inducible expression of a secreted protein.

Sequence Author: Thermo Fisher (Invitrogen)

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BglII (4689) MluI (4595) BsrGI (4588) PpuMI (4514) EcoRV (4421) StuI (4387) DraIII (4336) FseI (4259) CsiI - SexAI * (4146) SgrAI (4097) SmaI (4073) TspMI - XmaI (4071) MauBI (4019) MscI (3986) NcoI - StyI (3981) HaeII (3265) AlwNI (3096) AhdI (2617) BmrI (2577) FspI (2394) SphI (5) BbvCI (49) NsiI (284) NdeI (405) BspDI - ClaI (492) BstAPI (676) PsiI (817) PaeR7I - XhoI (865) BsaAI - PmlI (899) SfiI (909) Acc65I (925) KpnI (929) SacII (933) PspOMI (949) ApaI (953) SalI (1009) AccI (1010) 6xHis AgeI (1091) BsiWI (1223) BclI * (1239) PvuII (1257) BamHI (1372) AscI (1379) SspI (1812) XmnI (2017) PvuI (2248) pFLD α 4694 bp
BglII  (4689)
1 site
A G A T C T T C T A G A
MluI  (4595)
1 site
A C G C G T T G C G C A
BsrGI  (4588)
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.
PpuMI  (4514)
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.
EcoRV  (4421)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
StuI  (4387)
1 site
A G G C C T T C C G G A
DraIII  (4336)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
FseI  (4259)
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.
CsiI  (4146)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (4146)
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.
SgrAI  (4097)
1 site
C R C C