Bacterial expression plasmid for SpRY-HF1 with an n-terminal MKIEE MBP tag, and c-terminal bi-partite NLS and 6x-His tag

Vector backbonepET28

Vector typeBacterial Expression ; in vitro transcription; T7 promoter

GROWTH IN BACTERIA

Bacterial Resistance(s)Kanamycin, 50 μg/mL

Growth Temperature37°C

Growth Strain(s)DH5alpha

Copy numberLow Copy

GENE/INSERT

Gene/Insert namebacterial codon optimized SpRY-HF1

Alt namepMM-gbr2102

SpeciesSynthetic

MutationSpRY-HF1, SpRY=A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R; HF1=N497A/R661A/Q695A/Q926A

PromoterT7

Tags / Fusion Proteins

MKIEE MBP tag (N terminal on insert)

BPNLS(SV40)-6xHis (C terminal on insert)

CLONING INFORMATION

Cloning methodGibson Cloning

5′ sequencing primeroBK311-GCCATTCGATGGTGTCCGG

3′ sequencing primeroBK2433-GAGGCCCCAAGGGGTTATGCTAG

Map：