pVP68K

Bacterial Flexi® Vector with a kanamycin resistance marker, for appending an N-terminal 8xHis-MBP-HRV 3C cassette to an expressed protein.

Sequence Author: Center for Eukaryotic Structural Genomics

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FspI (5948) NaeI (5548) NgoMIV (5546) RsrII (5531) AvrII (5379) PciI (4558) NdeI (4381) BstZ17I (4331) BstEII (3525) EcoRV (3261) PaeR7I - XhoI (117) AanI (165) MfeI (175) lac operator RBS NcoI (229) BseRI (231) ATG NsiI (263) BsaI (327) AanI (616) BmgBI (806) AfeI (1174) PacI (1369) AsiSI - SgfI (1417) NotI (1420) Bpu10I (1871) PasI (2330) SspI (2410) ScaI (2515) PmeI (2549) Eco53kI (2562) SacI (2564) SbfI (2597) BsrGI (2703) AscI (2720) SgrDI (2735) BspDI - ClaI (2748) BstBI (2759) HindIII (2762) HpaI (3205) pVP68K 6353 bp
FspI  (5948)
1 site
T G C G C A A C G C G T
NaeI  (5548)
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  (5546)
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.
RsrII  (5531)
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.
AvrII  (5379)
1 site
C C T A G G G G A T C C
PciI  (4558)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NdeI  (4381)
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.
BstZ17I  (4331)
1 site
G T A T A C C A T A T G
BstEII  (3525)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
EcoRV  (3261)
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.
PaeR7I  (117)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (117)
1 site
C T C G A G G A G C T C
AanI  (165)
2 sites
T T A T A A A A T A T T
MfeI  (175)
1 site
C A A T T G G T T A A C
NcoI  (229)
1 site
C C A T G G G G T A C C