831 pMiniT3

Digest  832 pMiniT-VVTopo with EcoRV, religate and transform NEB Turbo

PCR screening colonies using pMiniT-seq-F and pMiniT-seq-R

831-pET28-F

ATCGGATCCGAATTCAGGAGGTAAAAACCATGATATCAGATGGCGCTGGGCGCAATG

831-pET28-R
CTGTTAATTAATTATTATCAGATCAGCTGTTTGATGGTGGTTAACGGCGGGA
(TCCCGCCGTTAACCACCATCAAA CAG CTG ATCTGATAATAATTAATTAA cag  )

PCR fragment from pET28b

ligate to pMiniT2.
positive clone: digest with BamH1 + PacI, religate to cut out insert